СПОСОБ ПОЛУЧЕНИЯ РЕБАУДИОЗИДА N С ПРИМЕНЕНИЕМ ФЕРМЕНТАТИВНОГО СПОСОБА

ШТАММ ГРИБА AUREOBASIDIUM PULLULANS - ПРОДУЦЕНТ ЭКЗОПОЛИСАХАРИДА

Изобретение относится к микробиологической промышленности и касается нового штамма гриба - продуцента внеклеточного полисахарида, предназначенного для изготовления различных лекарственных форм. Цель изобретения - получение штамма гриба Aureobasidium pullulans ВКПМ F-370 (лабораторный номер 16а), способного к повышенному синтезу экзополисахарида, отличающегося по химическому строению, молекулярной массе и другим свойствам от экзополисахаридов, синтезируемых известными штаммами. Полисахарид представляет собой глюкан, содержащий 21% - 1,3, 52% - 1,4 и 27% - 1,6 гликозидных связей. Вязкость 0,1%-ного водного раствора полисахарида 2,5, мол.м. около 1,3·106 Da . После 4 сут выращивания на среде с сахаром, сернокислым аммонием, минеральными солями и дрожжевым экстрактом образуется до 18,6 г/л нативного и 9,2 г/л чистого полисахарида. 3 табл.

Мутантная АТФ-зависимая протеаза и способ получения L-аминокислоты с ее применением

Изобретение относится к биотехнологии. Предложен полипептид, участвующий в получении аминокислоты с разветвленной цепью, содержащий делецию аминокислот, соответствующих положениям 431-433 на основании аминокислотной последовательности, представленной SEQ ID NO: 5, где аминокислота с разветвленной цепью представляет собой L-валин или L-изолейцин. Также предложен микроорганизм рода Corynebacterium для получения указанных аминокислот с разветвленной цепью, в котором активность АТФ(аденозинтрифосфат)-зависимой протеазы ослаблена по сравнению с немодифицированным микроорганизмом, где указанный микроорганизм содержит вышеуказанный полипептид или полинуклеотид, имеющий делецию нуклеотидов, соответствующих положениям 1291-1299 на основании последовательности, представленной SEQ ID NO: 6. Также предложены способ получения указанных аминокислот с разветвленной цепью с использованием указанного микроорганизма, а также применение микроорганизма или полипептида для получения указанных аминокислот с ...

Способ получения клинического декстрана

Verfahren zur Herstellung von Antitumordextran

WATER SOLUBLE IRON-DEXTRAN PREPARATION

Production and fractionation of dextrans

Addition products are obtained by adding alkaline earth hydroxides to solutions of dextran, and these may be decomposed by acids to give free dextran in solution and insoluble alkaline earth metal salts. The process is particularly applicable to the purification of dextran from fermentation residues containing it and to the fractionation of dextran and/or partially degraded dextrans of different molecular weight. The latter may be achieved either by fractional addition of alkaline earth hydroxide, the lower molecular weight complexes being precipitated first, or by fractional acidification of the precipitate when the higher molecular weight complexes are preferentially dissolved. The inorganic salts can be removed from the dextran solution by dialysis or adsorption on ion-exchange resins. When calcium hydroxide is added to fermentation residues containing dextran and fructose the precipitated complex contains both these but with strontium and barium only the dextran is precipitated. If ...

USE OF A GLUKAN FROM LACTIC ACID BACTERIA AS CORROSION PROTECTION AGENTS

PRODUCTION FROM DEXTRAN LEUCONOSTOC DEXTRANICUM

Polysaccharide fibers

GLUCO-OLIGOSACCHARIDE MIXTURE AND A PROCESS FOR ITS MANUFACTURE

METHOD FOR THE PRODUCTION OF DEXTRAN

Method for the production of dextran comprising the following steps: prepare a culture medium containing the appropriated mixture and balance of ingredients, mainly after accurate selection of nature and concentration of carbon and nitrogen sources, with a specific initial pH, inoculate the culture medium with an appropriated quantity of bacteria strain (to standardize the production and avoid as much as possible the variability of the system); carry out the fermentation for a given time and at a given temperature; precipitate the dextran to separate the product from the culture medium; the bacteria strain is a strain of Weissella cibaria.

GELLING DEXTRAN ETHERS

Compositions are disclosed herein comprising at least one dextran ether compound that comprises uncharged, anionic, and/or cationic organic groups. The degree of substitution of one or more dextran ether compounds is about 0.0025 to about 3Ø Dextran from which the disclosed ether compounds can be derived can have a weight-average molecular weight of about 50-200 million Daltons and/or a z-average radius of gyration of about 200-280 nm. Also disclosed are methods of producing dextran ether compounds, as well as methods of using these ether compounds in various applications.

ENZYMATIC HYDROLYSIS OF DISACCHARIDES AND OLIGOSACCHARIDES USING ALPHA-GLUCOSIDASE ENZYMES

A method is disclosed for hydro!yzing an alpha-1,5 giucosyl-fructose linkage in a saccharide (disaccharide or oligosaccharide) such as leucrose. This method comprises contacting the saccharide with an alpha-glucosidase enzyme such as transgiucosidase or glucoamylase under suitable conditions, during which contacting step the enzyme hydrolyzes at least one aipha-1,5 glucosyl-fructose linkage of the saccharide. This method is useful for reducing the amount of leucrose in a filtrate isolated from a glucan synthesis reaction, for example.

Verfahren zur Herstellung eines Dextran-Abbauproduktes für pharmazeutische Zwecke.

Verfahren zur Herstellung von kolloidal löslichen Dextranen

Procédé de production de dextrane utilisable à des fins thérapeutiques.

Verfahren zur Herstellung von Dextran aus Saccharose

Verfahren zur biologischen Herstellung von Dextranen, insbesondere für klinische Zwecke

Способ получения бета-циклодекстрина

Изобретение относится к биотехнологии, в частности, к способам получения микробиологическим синтезом бета-циклодекстринов. Штамм-продуцент циклодекстринглюконотрансферазы выращивают в жидкой среде, из культуральной жидкости выделяют фермент, который далее используют в качестве биокатализатора в реакции биотрансформации бета-циклодекстрина из крахмала. В качестве штамма-продуцента циклодекстринглюконотрансферазы используют штамм Bacillus halophilus BIO-12H. Обеспечивается повышение выхода бета-циклодекстрина в 1,5-2 раза.

Штамм бактериальной культуры LEUCONOSTOC MESENTEROIDES БМП-98 для производства декстрана

Штамм бактериальной культуры Leuconostoc mesenteroides БМП-98 (КПМКБ В18-1) для производства декстрана - субстанции плазмозамещающих препаратов.

An active fermented product and its preparation method and application

Yeast glucan production process

Containing β-1, 3-D-glucan extract and use thereof

A containing Candida utilis β - D - glucan of facial cleanser

Spatial hericium coralloides ST21-2 for producing polysaccharide and application thereof to improvement of biological oxidization resistance

A linkage producing high-purity β 1, 3-glucan preparation process

Method for preparing resistant dextrin through debranching and crystallization

The invention relates to the technical field of deep processing of starch, in particular to a method for preparing resistant dextrin by debranching and crystallization, which specifically comprises the following steps: gelatinizing starch at high temperature; secondly, pullulanase is used for enzymolysis to remove amylopectin, namely part of alpha-1, 4 glucosidic bonds in the starch are broken, new glucosidic bonds connected with alpha-1, 2 and alpha-1, 3 bonds and dextran, beta-1, 6 glucosidic bonds and the like formed by intramolecular dehydration on part of reducing ends are regenerated, and then the resistant dextrin has digestion resistance; further preparing resistant dextrin by adopting different debranching and crystallizing methods; and finally, carrying out enzyme hydrolysis by using porcine pancreas alpha-amylase to purify the resistant dextrin. The whole process is mild in condition, no chemical reagent is added, no wastewater is generated, the production process is pollution-free ...

CONSTRUCTION THE NEW VARIABLE ONES OF ENZYME DEXTRANE-SACCHARASE DSR-S BY MOLECULAR INGIENERIE.

Process for the manufacture of iso-malto-oligosaccharide monovalent haptens

Iso-malto-oligosaccharide monovalent haptens were prepared by adding an aqueous sucrose solution to an aqueous solution of D-glucose containing more than 300 mmol glucose per 1000 U α(1→6)-D-glucosyl transferase at 265 to 310 K and a pH value of from 4.5 to 8.0 and a molar ratio of sucrose to glucose of from 0.5 to 2.0. After consumption of the sucrose, glucose, liberated fructose and undesired oligosaccharides are separated in a known manner. The process of the invention allows a particularly economical preparation of the monovalent haptens which serve for the prophylaxis of undesired dextran induced anaphylactoid side effects (DIAR).

POLYPEPTIDE HAVING THE CAPACITY TO FORM ALPHA-1,3 GLUCOSYL UNIT BRANCHINGS ON AN ACCEPTOR

Polypeptides having the ability to specifically form connections of glucosyl units in alpha 1,3 on an acceptor having at least one hydroxyl moiety are presented. The polypeptides include i) the pattern I of sequence SEQ ID NO: 1, ii) the pattern II of sequence SEQ ID NO: 2, iii) the pattern III of sequence SEQ ID NO: 3, and iv) the pattern IV of sequence SEQ ID NO: 4, or derivates from one or several of said patterns, wherein the polypeptide furthermore has an aspartic residue (D) at position 5 of the pattern II (SEQ ID NO: 2), a glutamic acid residue (E) at position 6 of the pattern III (SEQ ID NO: 3) and an aspartic acid residue (D) at position 6 of the pattern IV (SEQ ID NO: 4). Methods for producing acceptors connected to glucosyl units in alpha 1,3 using the polypeptides are also provided.

A PROCESS FOR PRODUCING ALTERNAN-OLIGOSACCHARIDE

A process (S1) for producing alternan-oligosaccharide (8), comprising contacting in a reactor (11) sucrose (9) with a catalytically effective amount of alternansucrase enzyme (13) and acceptor molecules (12), wherein the alternansucrase enzyme (13) and acceptor molecules (12) are present in the reactor (11) in an aqueous liquid (4) and the sucrose (9) is continuously or half-continuously fed to the reactor (11), and wherein the sucrose (9) and the acceptor molecules (12) are converted to alternan-oligosaccharide (8), and fructose (6) is formed as a by-product, continuously or half-continuously removing at least a part of the fructose (6) from the reactor (11) by membrane filtration (17).

Process for producing polysaccharide ron substance

A biologically active polysaccharide RON substance which is excellent in such biological activities as antitumor activities, immunomodulating activities and host defense activities against infectious diseases, and process for producing said RON substance by treating sucrose with said RON substance synthetase.

BIOREACTOR WITH IMMOBILIZED LACTIC ACID BACTERIA AND THE USE THEREOF

MODIFIKATION VON POLYSACCHARIDEN

A process for the enzymatic production of dextran from saccharose

Dextran is produced from sucrose solutions of concentration 30 per cent or over by the action of an enzyme preparation made by culturing Leuconostoc meserentoides in a buffered dilute sucrose solution containing hydrolyzed protein nutrient, e.g. in a liquid containing 15 g. of sucrose, 10 g. of hydrolyzed protein and 5 g. of pulverized chalk per litre, and either adding ethyl alcohol to precipitate protein together with the enzymes and dextran so produced and dissolving the precipitate in a buffer solution of pH 5.5 or by adding tricalcium phosphate to adsorb protein together with the enzymes and then eluting the adsorbent with a buffer solution of pH 5.5.

Preparation and standardization of immunomodulatory peptide-linked glucans with verifiable oral absorbability from coriolus versicolor

IMPROVED PROCEDURE FOR THE PRODUCTION OF EXOZELLULAEREN BIO POLYMERS.

GLUCO OLIGOSACCHARID MIXTURE AND PROCEDURE FOR ITS PRODUCTION.

ALTERNANSUCRASE CODING NUCLEIC ACID OF MOLECULES

PROCEDURE FOR THE PRODUCTION OF AN IMPROVED STRUCTURAL STRUCTURE OF BAKING GOODS

NOVEL GLUCANS AND NOVEL GLUCANSUCRASES DERIVED FROM LACTIC ACID BACTERIA

Method for preparing a self-sufficient fermentation medium

ENZYME PRODUCTION AND PURIFICATION

Способ получения альфа-циклодекстрина

Изобретение относится к биотехнологии, в частности, к микробиологическим способам получения альфа-циклодекстринов. Штамм-продуцент циклодекстринглюконотрансферазы выращивают в жидкой питательной среде, из культуральной жидкости выделяют фермент, который используют в качестве биокатализатора в процессе трансформации альфа-циклодекстрина из крахмала. Трансформацию проводят при температуре 50-55°С, при отсутствии органических растворителей. В качестве источника биокатализатора используют штамм бактерий Thermoactinomyces vulgaris Act-8 (INMIA-3554). Достигается повышение выхода альфа-циклодекстрина в 1,5 раза.

Expression of dextran sucrase genetic engineering bacteria, construction method and use thereof

The invention relates to a genetic engineering strain with the expression of a dextransucrase, a dextransucrase gene (dexYG) is inserted in an expression plasmid pET28a to construct a recombinant expression plasmid pET28-dexYG, and the recombinant expression plasmid is converted into E.coli BL21(DE3), thereby obtaining the genetic engineering strain BL21(DE3)/pET28-dexYG with the high efficient expression of the dextransucrase. The engineering strain can carry out high-expression and stable fermentation for producing the dextransucrase under the appropriate induction conditions. Sucrose is taken as a substrate, the dextransucrase is used for producing dextran under the simpler catalytic conditions, and the molar conversation rate can achieve 70 to 85 percent. The genetic engineering strain can fundamentally upgrade the domestic prior dextran production process and obtain the dextran production process with advanced process and great quality.

Proceeded of preparation of products containing dextrane intend for pharmaceutical and therapeutic preparations

processo de obtenção de dextrana clínica, oligossacarídeos e frutose a partir da sacarose

Process for obtaining improved structure build-up of baked products

A process for obtaining improved structure build-up of baked products includes the steps of incorporating a sufficient amount of exopolysaccharides into a dough to show a rise in viscosity with time and thereafter maintaining the achieved viscosity. New dextrans and new micro-organisms producing them can be used in the process. Thus, the dough and baked products containing these dextrans are produced.

SYNTHESIS OF GLUCAN COMPRISING ALPHA-1,3 GLYCOSIDIC LINKAGES WITH PHOSPHORYLASE ENZYMES

Reaction compositions are disclosed herein comprising at least water, beta-glucose-1-phosphate (beta-G1P), an acceptor molecule, and an alpha-1,3-glucan phosphorylase enzyme. These reactions can synthesize oligosaccharides and polysaccharides with alpha-1,3 glycosidic linkages. Further disclosed are alpha-1,3-glucan phosphorylase enzymes and methods of use thereof.

ALPHA-1,3-GLUCAN GRAFT COPOLYMERS

Compositions are disclosed herein comprising a graft copolymer that comprises: (i) a backbone comprising dextran that has been modified with about 1%-25% alpha-1,2 branches, and (ii) one or more alpha-1,3-glucan side chains comprising at least about 50% alpha-1,3 glycosidic linkages. Further disclosed are reactions for producing such graft copolymers, as well as their use in derivatives, films and various other applications.

WASSERLOESLICHES EISENDEXTRAN AND PROCEDURES FOR ITS PRODUCTION.

Method of increasing the alpha-amylase-resistant starch content (rs content) of a polysaccharide, polysaccharides, the use thereof and food containing said polysaccharides

Enzymatic hydrolysis of disaccharides and oligosaccharides using alpha-glucosidase enzymes

A method is disclosed for hydro!yzing an alpha-1,5 giucosyl-fructose linkage in a saccharide (disaccharide or oligosaccharide) such as leucrose. This method comprises contacting the saccharide with an alpha-glucosidase enzyme such as transgiucosidase or glucoamylase under suitable conditions, during which contacting step the enzyme hydrolyzes at least one aipha-1,5 glucosyl-fructose linkage of the saccharide. This method is useful for reducing the amount of leucrose in a filtrate isolated from a glucan synthesis reaction, for example.

COMPOUNDS AND METHODS FOR DIAGNOSIS AND TREATMENT OF VIRAL INFECTIONS

Compositions and methods of using these compositions that can include a targeting moiety and a therapeutic agent are described herein. These compositions can be used for di-agnosing and/or treating flaviviridae-family viruses including Zika virus, dengue virus, and yellow fever.

ENZYMATIC SYNTHESIS OF SOLUBLE GLUCAN FIBER

An enzymatically produced soluble a-glucan fiber composition is provided suitable for use as a digestion resistant fiber in food and feed applications. The soluble a-glucan fiber composition can be blended with one or more additional food ingredients to produce fiber-containing compositions. Methods for the production and use of compositions comprising the soluble a-glucan fiber are also provided.

PREPARATION AND STANDARDIZATION OF IMMUNOMODULATORY PEPTIDE-LINKED GLUCANS WITH VERIFIABLE ORAL ABSORBABILITY FROM CORIOLUS VERSICOLOR

... ²²²This invention provides compositions and methods for stimulating the immune ²system. Such methods include administering an extract, purified peptide-linked ²glucan or active component thereof from Coriolus Versicolor. The methods are ²particularly useful for prophylactic and therapeutic treatment of secondary ²immunodeficiency, wherein the immunodeficiency is the result of an infection, ²a malignant neoplastic disease, an autoimmune disease, a protein losing state, ²an immunosuppressive treatment, surgery or anesthesia.² ...

MODIFICATION OF POLYSACCHARIDES

The present invention relates to a method for manufacturing modified polysaccharides comprising of placing the polysaccharide in contact with a sugar group-transferring enzyme and a sugar group donor. The placing of the polysaccharide in contact with a sugar group-transferring enzyme and a sugar group donor can be effected in vivo as well as in vitro. The result of the method is modified polysaccharides which can be used for different food and non-food applications.

Fermentation and separation process for obtaining great amount of extracellular mycose

Water-soluble yeast beta-dextran and preparation thereof

The invention relates to a water-soluble barm Beta-dextran and a preparation method thereof. In the water-soluble barm Beta-dextran chain, Beta-1, 3-dextran is taken as a main chain, Beta-1, 6-dextran is taken as a branched chain, the molecular weight is 0.08 to 0.2 million Daltun. The preparation method of the water-soluble barm Beta-dextran includes the following steps of: water extraction, alkali extraction and impurity processing. The preparation technique of the invention is simple; the technique parameters are easy to be operated and controlled; both the yield and the purity of the product water-soluble barm Beta-dextran are higher. The obtained water-soluble barm Beta-dextran has remarkable immune activation effect and especially can improve the phagocytic function and the antibodyformation capacity of a macrophage to a large extent, thereby improving the body immunity and resistance. The water-soluble barm Beta-dextran can be widely applied in the fields such as food, dairy food ...

Apparatus of vulcanization to autonomous heating

Dextran A method of manufacturing

Cement grout, container of the dextrane, with weak water loss

HYDROLYSIS METHOD, COMPOSITION AND METHODS OF ENRICHING FRUCTOSE AND FERMENTATION

POLYSIALIC ACID, BLOOD GROUP ANTIGENS AND GLYCOPROTEIN EXPRESSION

Process for obtaining improved structure build-up of baked products

A process for obtaining improved structure build-up of baked products includes the steps of incorporating a sufficient amount of exopolysaccharides into a dough to show a rise in viscosity with time and thereafter maintaining the achieved viscosity. New dextrans and new micro-organisms producing them can be used in the process. Thus, the dough and baked products containing these dextrans are produced.

PROTEIN WITH DEXTRAN-SACCHARASE ACTIVITY, AND USES

The invention relates to: a protein having dextran-saccharase activity and the sequence SEQ ID NO: 1 for an no acid sequence; a protein having dextran-saccharase activity and the sequence SEQ ID NO: 2 for an amino acid sequence; a complex containing a substrate and a protein with dextran-saccharase activity a method for synthesizing dextrans; and dextrans. The invention also relates to a method for synthesizing gluco-oligosaccharides and gluco-conjugates and to the resulting products.

ENZYMATIC HYDROLYSIS OF DISACCHARIDES AND OLIGOSACCHARIDES USING ALPHA-GLUCOSIDASE ENZYMES

RESISTANT STARCH

STARCH DECOMPOSITION PRODUCT, AND FOOD ADDITIVE, FOOD AND DRINK AND DRUG CONTAINING THE STARCH DECOMPOSITION PRODUCT, AND METHOD FOR PRODUCING THE STARCH DECOMPOSITION PRODUCT

PROBLEM TO BE SOLVED: To provide a new starch decomposition product showing a low viscosity, low sweet taste, and a low osmotic pressure, compared to an existing starch decomposition product showing the same DE value. SOLUTION: This starch decomposition product is such that a content of components with a glucose polymerization degree (DP) of 8-19 is ≥32%, and a content of components with a glucose polymerization degree (DP) of ≥20 is ≤30%. The starch decomposition product shows a low viscosity, low sweet taste, and a low osmotic pressure, compared to an existing starch decomposition product showing the same DE value. A method for producing the starch decomposition product, including at least a process of making debranching enzyme act on starch, and a process of making branching enzyme act on starch is also provided. COPYRIGHT: (C)2011,JPO&INPIT ...

СПОСОБ ПОЛУЧЕНИЯ РЕБАУДИОЗИДА N С ПРИМЕНЕНИЕМ ФЕРМЕНТАТИВНОГО СПОСОБА

Изобретение относится к биотехнологии. Предложен способ получения ребаудиозида N с применением ферментативного способа, включающий использование ребаудиозида А или ребаудиозида J в качестве субстрата и получение субстрата в присутствии донора гликозила в реакции в условиях катализа UDP-гликозилтрансферазой и/или содержащими UDP-гликозилтрансферазу рекомбинантными клетками. Способ позволяет получить ребаудиозид N высокой чистоты с низкой стоимостью в течение короткого производственного цикла. 11 з.п. ф-лы, 8 пр.

Штамм @ @ ЛУ-122-продуцент декстрана,обладающего иммуностимулирующим действием

Штамм Leuconostoc dextranicum Лу-122 ВКМ В1526Д (Всесоюзная коллекция микроорганизмов при Институте биохимии и физиологии микроорганизмов АН СССР) - продуцент декстрана, обладающего иммуностимулирующим действием .

BIOREAKTOR MIT IMMOBILISIERTEN MILCHSÄUREBAKTERIEN UND SEINE VERWENDUNG

Verfahren zur Herstellung monovalenter Haptene

Improvements in and relating to the production of dextran from sucrose by means of dextran-forming bacteria

In the process of producing dextran from sucrose-containing material by treatment with dextran-forming bacteria, synthetic p-amino-benzoic acid is added to the culture medium. The bacteria employed is preferably Betacoccus arabinosaceus.

PROCEDURE FOR THE PRODUCTION OF POLYSACCHARIDE ABSTENTION NEW ONE DRIED COMPOSITIONS.

PROCEDURE FOR THE PRODUCTION OF MONOVALENTER HAPTENE.

MODIFICATION OF POLYSACCHARIDEN

A method for synthesis of polysaccharides

ENZYMATIC SYNTHESIS OF SOLUBLE GLUCAN FIBER

An enzymatically produced soluble a-glucan fiber composition is provided suitable for use as a digestion resistant fiber in food and feed applications. The soluble a-glucan fiber composition can be blended with one or more additional food ingredients to produce fiber-containing compositions. Methods for the production and use of compositions comprising the soluble a-glucan fiber are also provided.

PROCESS FOR OBTAINING IMPROVED STRUCTURE BUILD-UP OF BAKED PRODUCTS

The present invention is related to a process for obtaining improved structure build-up of baked products comprising the steps of incorporating into a dough sufficient amount of exopolysaccharides, said exopolysaccharides showing in a solution a rise in viscosity with time and thereafter maintaining the achieved viscosity. The present invention is also related to new dextrans and new micro-organisms producing them, and the dough and baked products containing these dextrans.

Method for preparing yeast cell wall beta-1,3-dextran

The invention relates to a preparation method for yeast cytoderm beta-1,3-glucans, comprising the following steps: firstly, impurity treatment, namely yeast dry powder is dispersed into water and impurities in the yeast dry powder are filtered; secondly, alkali treatment, namely alkalis are added into filtered yeast suspension and final concentration of alkalis in the suspension is still 0.5 to 7 percent; thirdly, cleaning, namely supernatants are absorbed by means of siphonage until that the supernatants are clear without impurities and PH value is 7 to 8; fourthly, sponging drying, and beta-1,3-glucan solid powder is obtained finally. The invention has the advantages that: firstly, commercial process cost is reduced; secondly, because operating procedures and devices used are simple, the method is easy to realize commercial process; thirdly, protein removal rate in raw materials is high, and yield and purity of the product beta-1,3-glucans are high.

Method for recovering glucomannan and protein in konjac flying powder by enzyme process

The invention relates to a method for recovering glucomannan and protein in konjac flying powder by the enzyme process, which comprises the following process steps: (1) liquification: adding alpha-amylase into water solution of the konjac flying powder and liquifying till starch is fully degraded; (2) saccharification: adding amyloglucosidase into solution in the step (1) for saccharification; (3) centrifugation: regulating the pH of the solution in the step (2) to 3.5-5.0, centrifugating and getting precipitate; (4) acid washing: adding water into the precipitate in the step (3), regulating the pH of the solution to acidic condition, stirring, centrifugating, and getting the precipitate; and (5) drying: regulating the pH of the precipitate after the acid washing of the step (4) to be neutral, drying and finally getting a final extract. The process flow is simple, the operation is easy, the content of the glucomannan and crude protein in the got extract is high, and the extract can beused ...

Enzymatic process of production of the dextrane with parth of saccharose

CONSTRUCTION THE NEW VARIABLE ONES OF MOLECULAR ENZYME DEXTRANE-SACCHARASE DSR-S PARINGIENERIE.

La présente invention concerne un procédé recombinant pour la production de dextrane-saccharases tronquées et/ou mutées tout en conservant leur activité enzymatique. Plus précisément, la présente invention concerne des séquences d'acides nucléiques de dextrane-saccharases tronquées et/ou mutées, des vecteurs contenant de telles séquences d'acides nucléiques et des cellules hôtes transformées par les séquences encodant pour les dextrane-saccharases tronquées et/ou mutées. Selon un autre aspect de l'invention, celle-ci concerne un procédé pour produire de façon recombinante des dextrane-saccharases tronquées et/ou mutées qui conservent leur activité enzymatique ainsi que des procédés pour produire des dextranes de haute masse molaire aux propriétés rhéologiques modifiées par rapport à celles du dextrane synthétisé par l'enzyme native dans les mêmes conditions et des dextranes ou des isomaltooligosaccharides de masse molaire contrôlée. Les dextranes et IMO de l'invention peuvent être utilisés ...

Leuconostoc mesenteroides CJNU 0705 and use thereof

METHOD FOR PREPARING AN AGAROOLIGOSACCHARIDE WITHOUT DETERIORATION AND DISCOLORATION BY DISSOLVING AGAR AT LOW TEMPERATURE

PURPOSE: A method for preparing an agarooligosaccharide is provided to obtain the agarooligosaccharide easily having a desired molecular weight without deterioration or discoloration by dissolving agar at low temperature, improve the medicinal effect of the agar by maintaining a molecular weight in accordance with desired function and reduce the total manufacturing cost. CONSTITUTION: A method for preparing an agarooligosaccharide comprises the steps of: (a) dissolving agar in water at high temperature, thereby preventing the agar from being solidified even the temperature gets lowered; and (b) filtering and drying a reaction product after adding 0.5-10 wt.% of β-agarase to a solution obtained from the step(a) and allowing it to react for a certain period of time. © KIPO 2008 ...

COMPOUNDS AND METHODS FOR DIAGNOSIS AND TREATMENT OF VIRAL INFECTIONS

FOOD FOR DIABETICS

The invention relates to a carbohydrate mixture which is provided with at least one modified carbohydrate made of a base body and a carbohydrate residue coupled therewith. The base body is a digestable, glucose-containing carbohydrate in the form of a digestable glucan or a non-digestable storage carbohydrate, skeletal carbohydrate or low-molecular component thereof. The base body is coupled to a carbohydrate residue. Glucose release from the inventive carbohydrate mixture is thus reduced by at least 10 %, detected in an in-vivo digestion system based on pancreatine and compared to a carbohydrate mixture which contains the same amount of weight of non-modified carbohydrates. The postprandial blood glucose concentration increase after eating can be moderated by means of the inventive carbohydrate mixture. The glucose can thus be metabolised by diabetics in spite of the existing lack of insulin. The inventive carbohydrate mixture can be used in food for diabetics and in pharmaceuticals.

POLYSACCHARIDE FIBERS

L'invention concerne de nouvelles fibres obtenues à partir de polysaccharides 'alpha'(1};3), ainsi que leur processus de production. Les fibres de l'invention présentent des propriétés proches de celles du coton mais elles peuvent être produites sous forme de filaments continus tout au long de l'année. Ces fibres conviennent dans des applications textiles.

Modification of polysaccharides

The present invention relates to a method for manufacturing modified polysaccharide in contact with a sugar group transferring enzyme and a sugar group donor. The result of the method is modified polysaccharides, which can be used for different food and non-food applications.

NEW METHOD FOR MANUFACTURE OF DEXTRAN, DEXTRAN SOLUTION OBTAINED, AND USES

Extrakt von Peptid-verknüpften Glucanen und Peptid-verknüpfte Clucane mit nachweisbarer oraler Resorbierbarkeit aus Coriolus Versicolor und deren Reinigungsverfahren und Verwendungen

Gereinigter Extrakt von Coriolus versicolor, enthaltend mindestens ein Peptid-verknüpftes Glucan, das durch eine (1→3)-Bindung verknüpfte Glucosemoleküle umfasst, ein durchschnittliches Molekulargewicht von 0,7 kDa bis 3 kDa, bestimmt durch Größenausschluss-Chromatografie aufweist und immunstimulierende Aktivität hat.

PREPARATION AND PURIFICATION PROCESS

Hmo synthesis

The present invention relates to a cell to be stably cultured in a medium, which cell is adjusted for the production of oligosaccharides, the cell being transformed to comprise at least one nucleic acid sequence coding for an enzyme involved in oligosaccharide synthesis. In addition the cell is transformed to comprise at least one nucleic acid sequence coding for a protein of the sugar efflux transporter family, a functional homolog or derivative thereof. Further, the invention concerns a method for the production of oligosaccharides involving above cell.

Glycosyltransferase promoter

A glycosyltransferase promoter and a recombinant nucleic acid, plant cell and transgenic plant containing thereof are provided. The promoter includes a nucleotide sequence as set forth in any one of SEQ ID NOs: 1˜7, a fragment having at least 10 contiguous bases of any one of SEQ ID NOs: 1˜7 or a combination thereof, or a nucleotide sequence having 90% or more identity to the nucleotide sequence as set forth in any one of SEQ ID NOs: 1˜7.

Production of Multi-Antennary N-Glycan Structures in Plants

The invention provides methods for producing multi-antennary glycoproteins in plant and plant cells. In particular the invention provides plants comprising a chimeric gene comprising glucosaminyltransferase IV and plants comprising two chimeric genes comprising glucosaminyltransferase IV and V.

Production of galactosylated n-glycans in plants

The invention provides methods for increasing the levels of bi-antennary mono- and fully galactosylated N-glycans, and for decreasing the levels of hybrid-type galactosylated N-glycans on glycoproteins produced in plants or plant cells. In addition, the invention provides methods for the production of heterologous glycoproteins with increased levels of bi-antennary mono- and fully galactosylated N-glycans, or decreased levels of hybrid-type galactosylated N-glycans in plants or plant cells.

Enhanced citric acid production in aspergillus with inactivated asparagine-linked glycosylation protein 3 (alg3), and/or increased laea expression

Provided herein are fungi, such as Aspergillus niger , having a dolichyl-P-Man:Man(5)GlcNAc(2)-PP-dolichyl mannosyltransferase (Alg3) gene genetic inactivation, increased expression of a loss of aflR expression A (Lae), or both. In some examples, such mutants have several phenotypes, including an increased production of citric acid relative to the parental strain. Methods of using the disclosed fungi to make citric acid are also provided, as are compositions and kits including the disclosed fungi.

INCREASED POLY (ALPHA 1,3 GLUCAN) YIELD USING BORIC ACID

A process for production of poly(α1,3 glucan) from a renewable feedstock, for applications in fibers, films, and pulps. The effect of addition of boric acid in increasing the yield of the desired end products, poly(α1,3 glucan) and fructose, and decreasing formation of the undesired by-product leucrose. 2) The reaction solution of further comprising at least one primer.3) The reaction solution of wherein the primer is dextran.4) The reaction solution of wherein the primer is hydrolyzed poly(α1 claim 2 ,3 glucan).5) The reaction solution of wherein the primer is from any low to med molecular weight (340 Dalton-50 claim 2 ,000 Dalton) glucose-based carbohydrate.6) The reaction solution of wherein the primer is from any low to med (340 Dalton-50 claim 2 ,000 Dalton) non-glucose-based carbohydrate.7) The reaction solution of wherein the primer is from any combination of any low to med molecular weight glucose-based carbohydrate.8) The reaction solution of wherein the enzyme of (a) is a primer dependent enzyme.9) The reaction solution of wherein the primer is glucose.10) The reaction solution of wherein the enzyme of (a) is a primer independent enzyme.11) The reaction solution of wherein more than one enzyme of (a) is present in the reaction solution.12) The reaction solution of wherein one gtf enzyme is primer dependent and one gtf enzyme is a primer independent enzyme.13) The reaction solution of wherein the concentration of boric acid in the reaction solution is from about 100 millimolar to about 600 millimolar.14) The reaction solution of wherein the reaction pH is maintained from 6.5 to 8.1.15) The reaction solution of wherein the yield of poly(α1 claim 1 ,3 glucan) formed in the reaction solution improves from 0.08-0.1 g glucan/g sucrose to 0.25 g glucan/g sucrose.17) The process of wherein the yield of leucrose formed decreases from 44% sucrose to 4% of sucrose converted.18) The process of wherein the reaction pH is maintained from 6.5 to 8.0.19) The process of ...

INCREASED POLY (ALPHA 1,3 GLUCAN) YIELD USING TETRABORATE

A process for production of poly (α 1,3 glucan) from a renewable feedstock, for applications in fibers, films, and pulps. The effect of addition of tetraborate in increasing the yield of the desired end products, poly (α 1,3 glucan) and fructose, and decreasing formation of the undesired by-product leucrose. 2) The reaction solution of further comprising at least one primer.3) The reaction solution of wherein the primer is dextran.4) The reaction solution of wherein the primer is hydrolyzed poly (α 1 claim 2 ,3 glucan).5) The reaction solution of wherein the primer is from any low to med molecular weight (340 Dalton-50 claim 2 ,000 Dalton) glucose-based carbohydrate.6) The reaction solution of wherein the primer is from any low to med (340 Dalton-50 claim 2 ,000 Dalton) non-glucose-based carbohydrate.7) The reaction solution of wherein the primer is from any combination of any low to med molecular weight glucose-based carbohydrate.8) The reaction solution of wherein the enzyme of (a) is a primer dependent enzyme.9) The reaction solution of wherein the primer is glucose.10) The reaction solution of wherein the enzyme of (a) is a primer independent enzyme.11) The reaction solution of wherein more than one enzyme of (a) is present in the reaction solution.12) The reaction solution of wherein one gtf enzyme is primer dependent and one gtf enzyme is a primer independent enzyme.13) The reaction solution of wherein the concentration of tetraborate in the reaction solution is from about 100 millimolar to about 150 millimolar.14) The reaction solution of wherein the reaction pH is maintained from 6.0 to 7.75.15) The reaction solution of wherein the yield of poly (α 1 claim 1 ,3 glucan) formed in the reaction solution improves from 0.16 g glucan/g sucrose to 0.30 g glucan/g sucrose.17) The process of wherein the yield of leucrose formed decreases from 44% sucrose to 4% of sucrose converted.18) The process of wherein the reaction pH is maintained from 6.5 to 8.0.19) The ...

PmST2 ENZYME FOR CHEMOENZYMATIC SYNTHESIS OF ALPHA-2-3-SIALYLGLYCOLIPIDS

The present invention provides novel methods for preparing glycolipid products. Novel sialyltransferases are also disclosed. 1. A method of preparing a glycolipid product , the method comprising: SEQ ID NO:4 (PmST2),', {'sub': '6', 'SEQ ID NO:5 (PmST2-His), and'}, {'sub': '6', 'SEQ ID NO:6 (MBP-PmST2-His),'}], 'a) forming a reaction mixture comprising an acceptor glycolipid, a donor substrate comprising a sugar moiety and a nucleotide, and a polypeptide selected from the group consisting ofwherein the reaction mixture is formed under conditions sufficient to transfer the sugar moiety from the donor substrate to the acceptor glycolipid, thereby forming the glycolipid product.2. The method of claim 1 , wherein the acceptor glycolipid comprises a galactoside moiety.3. The method of claim 2 , wherein the galactoside moiety is selected from the group consisting of a β1-4 linked galactoside moiety and a β1-3 linked galactoside moiety.4. The method of claim 2 , wherein the acceptor glycolipid comprises a lactoside moiety or an N-acetyl lactoside moiety.5. The method of claim 2 , wherein the acceptor glycolipid comprises a Galβ1-3GlcNAc moiety or a Galβ1-3GalNAc moiety.6. The method of claim 1 , wherein the nucleotide sugar comprises a cytidine 5′-monophosphate (CMP)-sialic acid.7. The method of claim 6 , wherein the CMP-sialic acid comprises cytidine 5′-monophosphate N-acetylneuraminic acid (CMP-Neu5Ac) or a CMP-Neu5Ac analog.8. The method of claim 7 , further comprising:b) forming a reaction mixture comprising a CMP-sialic acid synthetase, cytidine triphosphate, and N-acetylneuraminic acid (Neu5Ac) or a Neu5Ac analog under conditions sufficient to form the CMP-Neu5Ac or CMP-Neu5Ac analog.9. The method of claim 8 , wherein steps a) and b) are performed in one pot.10. The method of claim 8 , further comprising:c) forming a reaction mixture comprising a sialic acid aldolase, pyruvic acid or derivatives thereof, and N-acetylmannosamine or derivatives thereof under conditions ...

FUCOSYL TRANSFERASE GENE

A DNA molecule is provided which comprises a sequence according to SEQ ID NO: 1 having an open reading frame from base pair 211 to base pair 1740 or having at least 50% homology to the above-indicated sequence, or hybridizing with the above-indicated sequence under stringent conditions, or comprising a sequence which has degenerated to the above-indicated DNA sequence because of the genetic code, the sequence coding for a plant protein having fucosyltransferase activity or being complementary thereto. 142.-. (canceled)43. A method of producing a recombinant glycoprotein , comprising expressing a recombinant glycoprotein in plants or plant cells , wherein an endogenous α1 ,3-fucosyltransferase production is suppressed or completely stopped ,wherein said endogenous α1,3-fucosyltransferase is identified by sequence comparison with the α1,3-fucosyltransferase sequence according to SEQ ID NO: 1 with an open reading frame from base pair 211 to base pair 1740, and at least suppressing said endogenous α1,3-fucosyltransferase production.441. The method of claim , wherein said endogenous α1 ,3-fucosyltransferase can be identified by sequence comparison with the α1 ,3-fucosyltransferase sequence according to SEQ ID NO: 1 with an open reading frame from base pair 211 to base pair 1740 by the program fastDB.451. The method according to claim , wherein the glycoprotein is a human protein.461. The method of claim , wherein the expression of the α1 ,3-fucosyltransferase is suppressed or completely blocked by a knock-out mutation of the endogenous α1 ,3-fucosyltransferase gene in said plant or plant cell.471. The method of claim , wherein the expression of the α-1 ,3-fucosyltransferase is suppressed or completely blocked by antisense inhibition in said plant or plant cell.481. The method of claim , wherein the expression of the α1 ,3-fucosyltransferase is suppressed or completely blocked by transfection with a polynucleotide comprising a sequence of at least 50 nucleotides which is ...

NOVEL FUCOSYLTRANSFERASES AND THEIR APPLICATIONS

The present invention relates to nucleic acid and amino acid sequences from and from , coding for/representing novel alpha-1,3-fucosyltransferases. The invention also provides uses and methods for using the alpha-1,3-fucosyltransferases to generate fucosylated products, such as oligosaccharides, (glyco)proteins, or (glyco)lipids, in particular of 3-fucosyllactose. 1. Isolated polynucleotide encoding a polypeptide with alpha-1 ,3-fucosyltransferase activity and comprising a nucleic acid sequence selected from the group consisting of:a) SEQ ID Nos. 1, 3, or 5 of the attached sequence listing;b) a nucleic acid sequence complementary to SEQ ID Nos. 1, 3, or 5;c) nucleic acid sequences which hybridize under stringent conditions to the nucleic acid sequences defined in a) and b) or their complementary strands.2. The isolated polynucleotide of consisting of the SEQ ID Nos. 1 claim 1 , 3 claim 1 , or 5 encoding a polypeptide with alpha-1 claim 1 ,3 fucosyltransferase activity.3. Vector claim 1 , containing a nucleic acid sequence as claimed in encoding a polypeptide with alpha-1 claim 1 ,3-fucosyltransferase activity claim 1 , the nucleic acid sequence being operably linked to control sequences recognized by a host cell transformed with the vector.4. An isolated polypeptide with alpha-1 claim 1 ,3-fucosyltransferase activity consisting of an amino acid sequence selected from the group consisting of:(a) an amino acid sequence shown in SEQ ID NO. 2, 4, or 6;b) an amino acid sequence of an allelic variant of an amino acid sequence shown in SEQ ID No. 2, 4, or 6, wherein said allelic variant is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID Nos. 1, 3, or 5;c) an amino acid sequence of an ortholog of an amino acid sequence shown in SEQ ID No. 2, 4, or 6, wherein said ortholog is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a ...

Alpha 1-3 n-galactosylstransferase with altered donor specificities, compositions and methods of use

The invention generally features compositions and methods based on the structure-based design of alpha 1-3 N-Acetylgalactosaminyltransferase (alpha 3 GalNAc-T) enzymes from alpha 1-3galactosyltransferase (a3Gal-T) that can transfer 2′-modified galactose from the corresponding UDP-derivatives due to substitutions that broaden the alpha 3Gal-T donor specificity and make the enzyme a3 GalNAc-T.

HIGH TITER PRODUCTION OF HIGHLY LINEAR POLY (ALPHA 1,3 GLUCAN)

A process for enzymatic preparation of a highly linear poly(α 1, 3 glucan) from sucrose is disclosed. The glucosyltransferase enzyme (gtfJ) from is used to convert sucrose to a highly linear poly(α 1, 3 glucan) in high titers. Hydrolyzed poly(α 1, 3 glucan) is used as the primer for the gtfJ enzyme reaction resulting in the formation of highly linear poly(α 1, 3 glucan). 1. A process for the synthesis of a highly linear poly(a 1 , 3 glucan) comprising an enzyme reaction solution comprising:a) sucrose;b) at least one glucosyltransferase enzyme; andc) at least one primer wherein said primer is hydrolyzed poly(α 1, 3 glucan);wherein said primer initiates the synthesis of said highly linear poly(α 1,3 glucan) through the action of the glucosyltransferase enzyme on the sucrose.2. The process of wherein the novel poly(α 1 claim 1 , 3 glucan) formed does not contain branches in its structure.3Streptococcus salivarius. The process of wherein the glucosyltransferase enzyme of is gtfJ.49. The process if wherein the at least one glucosyltransferase enzyme is a primer-independent enzyme.5. The process of wherein the at least one glucosyltransferase enzyme is a primer-dependent enzyme.6. The process of wherein more than one glucosyltransferase enzyme is present in the enzyme reaction solution.7. The process of wherein the more than one glucosyltransferase enzyme comprises a mixture of at least one primer-dependent enzyme and at least one primer-independent enzyme.8. A process for producing a highly linear poly(α 1 claim 6 , 3 glucan) in a reaction system comprising two chambers claim 6 , separated by a semi-permeable membrane claim 6 , wherein: i) sucrose;', 'ii) at least one glucosyltransferase enzyme; and', 'iii) at least one primer wherein said primer is hydrolyzed poly(α 1, 3 glucan); and, 'a) a first chamber comprises an enzyme reaction solution comprisingb) a second chamber, separated from the first chamber by a semi-permeable membrane in contact with the enzyme reaction ...

HIGH TITER PRODUCTION OF POLY (ALPHA 1,3 GLUCAN)

A process for enzymatic preparation of poly (α1, 3 glucan) from sucrose is disclosed. The glucosyltransferase enzyme (gtfJ) from is used to convert sucrose to fructose and poly (α1, 3 glucan). Application of semi-permeable membranes to continuously remove fructose, a by-product of the gtf enzyme, thus increasing the poly (α1, 3 glucan) liter, is disclosed. 1. A process for producing poly (α1 , 3 glucan) in a reaction system comprising two chambers , separated by a semi-permeable membrane , wherein: i) sucrose; and', 'ii) at least one glucosyltransferase enzyme; and, 'a) a first chamber comprises an enzyme reaction solution comprisingb) a second chamber, separated from the first chamber by a semi-permeable membrane in contact with the enzyme reaction solution wherein the semi-permeable membrane is permeable to fructose but impermeable to poly (α1, 3 glucan), facilitates continuous removal of fructose and other low molecular weight moieties while retaining poly (α1, 3 glucan) and the at least one glucosyltransferase enzyme inside the first chamber.2. The process of further comprising at least one primer.3. The process if wherein the glucosyltransferase enzyme is a primer-independent enzyme.4. The process of wherein the glucosyltransferase enzyme is a primer-dependent enzyme.5. The process of wherein the semi-permeable membrane facilitates accumulation of poly (α1 claim 1 , 3 glucan) to a concentration ranging from 30 grams per liter to 200 grams per liter.6. The process of wherein the semi-permeable membrane has a molecular weight cut-off from 12 claim 5 ,000 to 100 claim 5 ,000 Daltons.7. The process of wherein the semi-permeable membrane is a dialysis tubing.8Streptococcus salivarius.. The process of wherein the glucosyltransferase enzyme is gtfJ from9. The process of wherein the at least one primer is dextran.10. The process of wherein more than one glucosyltransferase enzyme is present in the enzyme reaction solution.11. The process of wherein the more than one ...

CHEMOENZYMATIC SYNTHESIS OF STRUCTURALLY HOMOGENEOUS ULTRA-LOW MOLECULAR WEIGHT HEPARINS

Methods for preparing synthetic heparins are provided. Synthetic heparin compounds, including ultralow molecular weight heparin compounds are provided. Also provided are methods of chemoenzymatically synthesizing structurally homogeneous ultra-low molecular weight heparins. Heparin compounds provided herein can have anticoagulant activity.

TREHALOSE PHOSPHORYLASES AND THEIR USE IN THE BIOCATALYTIC PRODUCTION OF TREHALOSE-ANALOGUES AND GLYCOSYL PHOSPHATES

The present invention relates to trehalose phosphorylases which are useful for the industrial production of trehalose-analogues and glycosyl phosphates. More specifically, the invention discloses trehalose phosphorylases which are mutated in specific amino acid regions. These specific mutations result in modified substrate specificities of the enzymes. In addition, the present invention discloses a wild type trehalose phosphorylase from the marine organism , and mutated types thereof, which are highly thermostable and have a broad acceptor and donor specificity. 1. A mutated trehalose phosphorylase containing at least one mutation in the amino acid positions 371 , 442 , 450 , 649 , 659 or 693 wherein said amino acid positions correspond to the amino acid positions as determined by SEQ ID N° 1 or to corresponding amino acid positions in a trehalose phosphorylase having an amino acid sequence which is at least 75% identical to SEQ ID N° 1 , and wherein said mutated trehalose phosphorylase has an increased catalytic efficiency towards its acceptor or donor substrate compared to the corresponding wild-type trehalose phosphorylase.2. The mutated trehalose phosphorylase according to wherein said mutation is a substitution at amino acid positions 371 claim 1 , 442 claim 1 , 450 claim 1 , 649 claim 1 , 659 and 693 of SEQ ID N° 1 or at corresponding amino acid positions in a trehalose phosphorylase having an amino acid sequence which is at least 75% identical to SEQ ID N° 1.36-. (canceled)7. The mutated trehalose phosphorylase of claim 1 , wherein said trehalose phosphorylase has a mutation at amino acid positions 442 claim 1 , 450 or 659 of SEQ ID N° 1.810-. (canceled)11. A mutated trehalose phosphorylase according to which is recombinantly expressed in a host cell.1213-. (canceled)14. A method to produce trehalose-analogues comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'contacting a mutated trehalose phosphorylase according to with β-glucose-1-phosphate and ...

METHODS FOR ALTERING THE REACTIVITY OF PLANT CELL WALLS

Methods and means are provided for the modification of the reactivity of plant cell walls, particularly as they can be found in natural fibers of fiber producing plants by inclusion of positively charged oligosaccharides or polysaccharides into the cell wall. This can be conveniently achieved by expressing a chimeric gene encoding an N-acetylglucosamine transferase, particularly an N-acetylglucosamine transferase, capable of being targeted to the membranes of the Golgi apparatus in cells of a plant. 1. A method for increasing the amount of positively charged oligosaccharides or polysaccharides in the cell wall , particularly the secondary cell wall of a plant cell , said method comprising 1. a plant-expressible promoter;', '2. a DNA region coding for an N-acetylglucosamine transferase, wherein said N-acetylglucosamine transferase can be targeted to the membranes of the Golgi-apparatus; and', '3. a transcription termination and polyadenylation region., 'i. Introducing or providing a chimeric gene to the plant cell, said chimeric gene comprising2ArabidopsisArabidopsis thaliana. The method of claim 1 , wherein said N-acetylglucosamine transferase comprises a signal anchor sequence selected from the signal anchor sequence of a rat sialyl transferase claim 1 , the signal anchor sequence of a human galactosyl transferase claim 1 , the signal anchor sequence of the homologue of the yeast HDEL receptor (AtERD2) claim 1 , the signal anchor sequence of the α-2 claim 1 ,6-sialyltransferase claim 1 , the signal anchor sequence of β1 claim 1 ,2-xylosyltransferase from claim 1 , the signal anchor sequence of N-acetylgluosoaminyl transferase I from tobacco or the amino acid sequence YYHDL or LKLEI.3. The method of claim 2 , wherein said N-acetylglucosamine transferase comprises the amino acid sequence of SEQ ID No. 15 from position 1 to position 35.4. The method of or claim 2 , wherein said N-acetylglucosamine transferase comprises the amino acid sequence of SEQ ID No. 15.5. The ...

THERMOPHILIC AND THERMOACIDOPHILIC BIOPOLYMER-DEGRADING GENES AND ENZYMES FROM ALICYCLOBACILLUS ACIDOCALDARIUS AND RELATED ORGANISMS, METHODS

Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from are provided. Further provided are methods of at least partially degrading, cleaving, or removing polysaccharides, lignocellulose, cellulose, hemicellulose, lignin, starch, chitin, polyhydroxybutyrate, heteroxylans, glycosides, xylan-, glucan-, galactan-, or mannan-decorating groups using isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from 1. An isolated or purified nucleic acid sequence comprising a nucleic acid sequence encoding a polypeptide selected from the group consisting of polypeptides having at least 90% sequence identity to SEQ ID No. 2 , 19 , 52 , 69 , 86 , 102 , 119 , 136 , 153 , 168 , 185 , 202 , 219 , 236 , 253 , 270 , 287 , 304 , 321 , 337 , 354 , 371 , 388 , 405 , 422 , and 439; at least 93% sequence identity to SEQ ID No. 462; at least 94% sequence identity to SEQ ID No. 36; at least 96% sequence identity to SEQ ID No. 460; at least 99% sequence identity to SEQ ID No. 464; at least 99.6% sequence identity to SEQ ID No. 458; and at least 99.7% sequence identity to SEQ ID No. 456.2. The isolated or purified nucleic acid sequence of claim 1 , wherein the polypeptide has enzymatic activity at or below about pH 7.3. The isolated or purified nucleic acid sequence of claim 1 , wherein the polypeptide has enzymatic activity at a temperature at or above about 50 degrees Celsius.4. The isolated or purified nucleic acid sequence of claim 1 , wherein the nucleic acid sequence is present in a vector.5. An isolated or purified polypeptide comprising a polypeptide selected from the group consisting of polypeptide having at least 90% sequence identity to SEQ ID No. 2 claim 1 , 19 claim 1 , 52 claim 1 , 69 claim 1 , 86 claim 1 , 102 claim 1 , 119 claim 1 , 136 claim 1 , 153 claim 1 , 168 claim 1 , 185 claim 1 , 202 claim 1 , 219 claim 1 , 236 claim 1 , 253 claim 1 , 270 claim 1 , 287 claim 1 , 304 claim 1 , 321 claim 1 , 337 claim 1 , ...

Method for industrially producing cyclic-structure-containing branched glucan

An object of the present invention is to provide a method for industrially producing a branched glucan having a cyclic structure. The method for producing a branched glucan having a cyclic structure comprises the steps of: (1) preparing a mixed liquid which contains a branching enzyme in which starch granules are suspended at a concentration of 5% by weight or more and 50% by weight or less, and allowing the branching enzyme to act on starch in the starch granules, wherein a temperature of the mixed liquid at the time of preparation is 0° C. or higher and not higher than the gelatinization starting temperature of the starch granule; and (2) elevating the temperature of the mixed liquid to 85° C. or higher and 129° C. or lower, wherein in the method, none of α-amylase, β-amylase, amyloglucosidase and a transglucosidase is added to the mixed liquid.

COMPOSITIONS RELATING TO A MUTANT CLOSTRIDIUM DIFFICILE TOXIN AND METHODS THEREOF

In one aspect, the invention relates to an immunogenic composition that includes a mutant toxin A and/or a mutant toxin B. Each mutant toxin includes a glucosyltransferase domain having at least one mutation and a cysteine protease domain having at least one mutation, relative to the corresponding wild-type toxin. The mutant toxins may further include at least one amino acid that is chemically crosslinked. In another aspect, the invention relates to antibodies or binding fragments thereof that binds to said immunogenic compositions. In further aspects, the invention relates to isolated nucleotide sequences that encode any of the foregoing, and methods of use of any of the foregoing compositions. 1. A composition comprising a polypeptide comprising SEQ ID NO: 84.2. The composition according to claim 1 , wherein the polypeptide comprises SEQ ID NO: 4.3. The composition according to claim 2 , wherein the methionine residue at position 1 is optionally not present.4. The composition according to claim 1 , wherein the polypeptide further comprises a beta-alanine moiety crosslinked to a side chain of a lysine residue of the polypeptide.5. The composition according to claim 4 , wherein the polypeptide further comprises a crosslink between a side chain of a second lysine residue of the polypeptide and a side chain of an aspartic acid residue of the polypeptide.6. The composition according to claim 4 , wherein the polypeptide further comprises a crosslink between a second lysine residue of the polypeptide and a side chain of a glutamic acid residue of the polypeptide.7. The composition according to claim 1 , wherein the polypeptide further comprises a crosslink between a side chain of a lysine residue of the polypeptide and a side chain of an aspartic acid residue of the polypeptide.8. The composition according to claim 1 , wherein the polypeptide further comprises a crosslink between a lysine residue of the polypeptide and a side chain of a glutamic acid residue of the ...

Fusion Enzymes

The present disclosure relates to recombinant proteins having N-acetylglucosaminyltransferase activity. The present disclosure further relates to methods for producing complex N-glycans including the steps of providing host cells containing such recombinant proteins and culturing the host cells such that the recombinant proteins are expressed. 1. A recombinant protein having N-acetylglucosaminyltransferase activity , wherein the recombinant protein catalyzes the transfer of N-acetylglucosamine to a terminal Manα3 residue and catalyzes the transfer of N-acetylglucosamine to a terminal Manα6 residue of an acceptor glycan , and wherein the recombinant protein comprises a catalytic domain from at least two different enzymes.2. The recombinant protein of claim 1 , wherein the recombinant protein is a fusion protein comprising an N-acetylglucosaminyltransferase I catalytic domain and an N-acetylglucosaminyltransferase II catalytic domain.3. The recombinant protein of claim 2 , wherein the N-acetylglucosaminyltransferase I catalytic domain and the N-acetylglucosaminyltransferase II catalytic domain are from human enzymes.4. The recombinant protein of claim 3 , wherein the N-acetylglucosaminyltransferase I catalytic domain comprises a sequence that is at least 70% claim 3 , at least 75% claim 3 , at least 80% claim 3 , at least 85% claim 3 , at least 90% claim 3 , at least 95% claim 3 , at least 96% claim 3 , at least 97% claim 3 , at least 98% claim 3 , at least 99% claim 3 , or 100% identical to amino acid residues 105-445 of SEQ ID NO: 1.5. The recombinant protein of claim 3 , wherein the N-acetylglucosaminyltransferase II catalytic domain comprises a sequence that is at least 70% claim 3 , at least 75% claim 3 , at least 80% claim 3 , at least 85% claim 3 , at least 90% claim 3 , at least 95% claim 3 , at least 96% claim 3 , at least 97% claim 3 , at least 98% claim 3 , at least 99% claim 3 , or 100% identical amino acid residues 30-447 of SEQ ID NO: 21.6. The ...

GNTIII EXPRESSION IN PLANTS

The invention relates to the field of glycoprotein processing in transgenic plants used as cost efficient and contamination safe factories for the production of recombinant biopharmaceutical proteins or pharmaceutical compositions comprising these glycoproteins. The invention provides a plant comprising a functional mammalian enzyme providing mammalian GnTIII that is normally not present in plants, said plant additionally comprising at least a second mammalian protein or functional fragment thereof that is normally not present in plants. 154.-. (canceled)55. A hybrid protein comprising:(a) an active domain of a GlcNAc-transferase III (GnTIII), and(b) a transmembrane region (Tm) of a protein that resides in the endoplasmic reticulum (ER) or Golgi apparatus of a eukaryotic cell or an ER retention signal peptide.56. The hybrid protein of claim 55 , wherein the protein that resides in the ER or Golgi apparatus of a eukaryotic cell is a glycosyltransferase.57. The hybrid protein of claim 56 , wherein the glycosyltransferase is selected from the group consisting of mannosidase I claim 56 , mannosidase II claim 56 , GlcNAc-transferase I claim 56 , GlcNAc-transferase II claim 56 , xylosyltransferase claim 56 , and fucosyltransferase.58. The hybrid protein of claim 56 , wherein the glycosyltransferase is a plant glycosyltransferase.59. The hybrid protein of claim 57 , wherein the hybrid protein is TmGnTI-GnTIII claim 57 , TmManII-GnTIII claim 57 , or TmXyl-GnTIII.60. The hybrid protein of claim 55 , wherein the ER retention signal peptide is KDEL (SEQ ID NO:28).61. The hybrid protein of claim 55 , wherein the GnTIII is a human GnTIII.62. The hybrid protein of claim 60 , wherein the human GnTIII comprises the amino acid sequence of SEQ ID NO:2.63. A plant host system claim 55 , which expresses the hybrid protein of .64. The plant host system of claim 63 , wherein the system is a plant cell.65. The plant host system of claim 63 , wherein the system is a whole plant.66. The ...

MALTOTRIOSYL TRANSFERASE, PROCESS FOR PRODUCTION THEREOF, AND USE THEREOF

The object is to provide a novel glycosyltransferase and the use thereof, the glycosyltransferase catalyzes transglucosylation of maltotriose units under conditions which can be employed for the processing of foods or the like. Provided is a maltotriosyl transferase which acts on polysaccharides and oligosaccharides having α-1,4 glucoside bonds, and has activity for transferring maltotriose units to saccharides, the maltotriosyl transferase acting on maltotetraose as substrate to give a ratio between the maltoheptaose production rate and maltotriose production rate of 9:1 to 10:0 at any substrate concentration ranging from 0.67 to 70% (w/v). 119-. (canceled)20. A maltotriosyl transferase gene comprising any of the DNAs selected from the group consisting of the following (a) to (e):(a) DNA coding the amino acid sequence set forth in SEQ ID NO: 7 or 8;(b) DNA comprising the sequence set forth in SEQ ID NO: 6;(c) DNA hybridizing with the complementary sequence of the sequence set forth in SEQ ID NO: 6 under stringent conditions;(d) DNA which is a degenerate of the DNA sequence of the sequence set forth in SEQ ID NO: 6;(e) DNA coding comprising a sequence including substitution, deletion, insertion, addition, or inversion of one or a plurality of bases with reference to the sequence set forth in SEQ ID NO: 6, and coding a protein having maltotriosyl transferase activity.21. A recombinant vector comprising the maltotriosyl transferase gene of .22. The recombinant vector of claim 21 , which is an expression vector.23. A transformant into which the maltotriosyl transferase gene of has been introduced.24. A transformant into which the recombinant vector of has been introduced.25. The transformant according to claim 23 , which is a bacterial cell claim 23 , a yeast cell claim 23 , or a fungal cell.26. A method for producing a maltotriosyl transferase claim 23 , comprising the following steps (i) and (ii):{'claim-ref': {'@idref': 'CLM-00023', 'claim 23'}, '(i) culturing the ...

Method for utilizing monoterpene glycosyltransferase

The object of the present invention is to provide a novel method for producing a terpene 8-glycoside. The present invention provides a method for producing a terpene 8-glycoside by means of glycosyltransferase acting on the 8-position of terpenes. The present invention provides a transformant transformed with a gene for the glycosyltransferase acting on the 8-position of terpenes and a method for producing such a transformant. The present invention provides a plant modified to suppress the expression of a protein having glycosylation activity on the 8-position of a monoterpene compound.

NOVEL USE OF MALTOTRIOSYL TRANSFERASE

The present invention is intended to provide a novel use of a maltotriosyl transferase. The present invention provides a method for producing rice cakes or noodles, including a step of heat-treating a dough containing maltotriosyl transferase thereby gelatinizing starch in the dough. The present invention also provides a method for producing an indigestible saccharide, including a step of allowing maltotriosyl transferase to act on a saccharide. 1. A method for producing rice cakes or noodles , comprising a step of heat-treating a dough containing maltotriosyl transferase thereby gelatinizing the starch in the dough.2. The production method according to claim 1 , wherein the dough is obtained by kneading a raw material powder which has been mixed with the maltotriosyl transferase claim 1 , or obtained by adding the maltotriosyl transferase during kneading a raw material powder.3. The production method according to claim 1 , wherein the main ingredient of the dough is rice flour claim 1 , glutinous rice claim 1 , wheat claim 1 , foxtail millet claim 1 , Japanese millet claim 1 , common millet claim 1 , buckwheat claim 1 , aroid claim 1 , sweet potato claim 1 , potato claim 1 , common sorghum claim 1 , corn claim 1 , green bean claim 1 , or soybean.4. The production method according to claim 1 , wherein the dough contains a pH controlling agent.5. The production method according to claim 1 , wherein the heat treatment step is achieved by steaming.6. A method for producing an indigestible saccharide claim 1 , comprising a step of allowing maltotriosyl transferase to act on a saccharide.7. The production method according to claim 6 , wherein the step is carried out using one or more enzymes selected from the group consisting of cyclodextrin glucanotransferase claim 6 , α-amylase (only those having transglycosylation activity) claim 6 , pullulanase claim 6 , and isoamylase in combination with the maltotriosyl transferase.8. The production method according to claim 6 , ...

Activated Sugars

Kinase and nucleotidyltransferase enzymes for the production of activated sugars have been developed. These enzymes have improved stability for industrial application and relaxed specificity towards a variety of sugars. These enzymes are useful in, for example, the production of diverse NDP-sugars for glycosylation of aglycones of interest, production of oligosaccharides, production of other important glycosylated sugars, and in drug discovery applications. 1. An isolated sugar-1-kinase , wherein the isolated sugar-1-kinase has sugar-1-kinase activity in a sugar-1-kinase assay and has a Thalf-life at 30° C. of greater than 10 minutes.2. The isolated sugar-1-kinase of claim 1 , wherein the sugar-1-kinase assay is a 3 claim 1 ,5-dinitrosalicylic acid (DNS) assay claim 1 , a thin layer chromatography assay or a high-performance liquid chromatography assay.3. The isolated sugar-1-kinase of claim 1 , comprising at least 90% amino acid sequence identity to SEQ ID NO:12 claim 1 , SEQ ID NO:8 claim 1 , SEQ ID NO:9 claim 1 , or SEQ ID NO:10 claim 1 , wherein the isolated sugar-1-kinase has sugar-1-kinase activity in a 3 claim 1 ,5-dinitrosalicylic acid (DNS) assay.4. The isolated sugar-1-kinase of claim 3 , comprising: (i) N120S; D183E; T191S; Y376F; and T381S;', '(ii) E71D and VI991;', '(iii) D221G; or', '(iv) a combination of one or more of the following mutations: N120S; D183E; T191S; Y376F; T381S; E71D; VI991; D221G; I341T; I341L, F375P F375M; F375Y; Y376K; Y376T; Y376P; and Y376F;, '(a) SEQ ID NO:8 with the following mutations (i) N119H; K130N; S239G; F238Y; and I312L;', '(ii) I312T and L332H;', '(iii) Y341P and F342K;', '(iv) Y341M and F342T;', '(v) I312T; L332H; Y341P; and F342K; or', '(vi) a combination of one or more of the following mutations: N119H; K130N; S239G; F238Y; I312L; I312T; L332H; Y341P; F342K; and Y341M; F342T; T168S; Y341P; Y341M; Y341F; F342K; F342T; F342P; F342Y;, '(b) SEQ ID NO:10 with the following mutations(c) SEQ ID NO:9 with the following ...

NOVEL FUCOSYLTRANSFERASES AND THEIR APPLICATIONS

The present invention relates to nucleic acid and amino acid sequences from serogroup O126, coding for/representing a novel alpha-1,2-fucosyltransferase. The invention also provides uses and methods for using the alpha-1,2-fucosyltransferase to generate fucosylated products, such as oligosaccharides, (glyco)proteins, or (glyco)lipids, in particular oligosaccharides found in human milk, such as 2′-fucosyllactose. 1. An isolated host cell for the production of a fucosylated oligosaccharide , the host cell comprising a nucleic acid sequence consisting of a polynucleotide encoding a polypeptide with alpha-1 ,2-fucosyltransferase activity and comprising a sequence selected from the group consisting of , a) the nucleic acid sequence shown in SEQ ID NO: 1 , b) a nucleic acid sequence complementary to SEQ ID NO: 1 , or c) nucleic acid sequences which hybridize under stringent conditions to the nucleic acid sequences defined in a) and b) or their complementary strands , wherein the nucleic acid sequence is a sequence foreign to the host cell and wherein the nucleic acid sequence is integrated in the genome of the host cell , or containing a vector comprising said nucleic acid sequence , wherein the nucleic acid sequence being operably linked to control sequences recognized by a host cell transformed with the vector.2. The isolated host cell according to claim 1 , wherein the host cell is selected from the group consisting of fungi including yeast claim 1 , bacteria claim 1 , insect claim 1 , animal and plant cells.3Escherichia coli. The isolated host cell according to claim 1 , wherein the host cell is an cell.4. The isolated host cell according to claim 1 , wherein the nucleic acid sequence consisting of the polypeptide encoding the polypeptide with alpha-1 claim 1 ,2-fucosyltransferase activity is adapted to the codon usage of the respective cell.5. A method for producing fucosylated oligosaccharides comprising the steps of:a) providing a polypeptide with alpha-1,2- ...

NOVEL GLYCOSYLTRANSFERASE GENE AND USE THEREOF

Provided is a polynucleotide encoding a protein having an activity to transfer a sugar to the hydroxy groups at the 4′- and 7-positions of a flavone. The polynucleotide is selected from the group consisting of: (a) a polynucleotide which comprises a base sequence represented by SEQ ID NO: 1, 3, or 12; (b) a polynucleotide which hybridizes to a polynucleotide comprising a base sequence complementary to a base sequence represented by SEQ ID NO: 1, 3, or 12 under high stringency conditions, and encodes a protein having an activity to transfer a sugar to the hydroxy groups at the 4′- and 7-positions of a flavone; (c) a polynucleotide which encodes a protein comprising an amino acid sequence represented by SEQ ID NO: 2, 4, or 13; (d) a polynucleotide which encodes a protein comprising an amino acid sequence in which one or more amino acids have been deleted, substituted, inserted, and/or added in an amino acid sequence represented by SEQ ID NO: 2, 4, or 13 and having an activity to transfer a sugar to the hydroxy groups at the 4′- and 7-positions of a flavone; etc. 1. A polynucleotide selected from the group consisting of:(a) a polynucleotide comprising a base sequence defined in SEQ ID NO: 1, 3 or 12;(b) a polynucleotide which hybridizes with a polynucleotide comprising a base sequence complementary to a base sequence defined in SEQ ID NO: 1, 3 or 12 under a stringent condition and encodes a protein having an activity of transferring a glycosyl to both of the hydroxyl groups at 4′- and 7-positions of a flavone;(c) a polynucleotide which encodes a protein comprising an amino acid sequence defined in SEQ ID NO: 2, 4 or 13;(d) a polynucleotide which encodes a protein comprising an amino acid sequence in which one or several amino acids have been deleted, substituted, inserted, and/or added in an amino acid sequence defined in SEQ ID NO: 2, 4 or 13 and having an activity of transferring a glycosyl to both of the hydroxyl groups at 4′- and 7-positions of a flavone; and(e) a ...

HIGH LEVEL EXPRESSION OF RECOMBINANT TOXIN PROTEINS

The present invention relates to the field of recombinant toxin protein production in bacterial hosts. In particular, the present invention relates to production processes for obtaining high levels of a recombinant CRM197, Diphtheria Toxin, Pertussis Toxin, Tetanus Toxoid Fragment C, Cholera Toxin B, Cholera holotoxin, and Exotoxin A, from a bacterial host. 1. A method for producing a recombinant toxin protein in a Pseudomonad host cell , said method comprising:ligating into an expression vector a nucleotide sequence encoding the toxin protein;transforming the Pseudomonad host cell with the expression vector; and {'i': 'C. difficile', 'wherein the recombinant toxin protein is Toxin B.'}, 'culturing the transformed Pseudomonad host cell in a culture media suitable for the expression of the recombinant toxin protein;'}2. The method of claim 1 , wherein the recombinant protein is produced at a yield of soluble and/or active toxin protein of 0.2 grams per liter to about 12 grams per liter.3. The method of claim 2 , wherein the yield of soluble and/or active toxin protein is about 0.2 grams per liter to about 12 grams per liter is about 0.2 g/L claim 2 , about 0.3 g/L claim 2 , about 0.4 g/L claim 2 , about 0.5 g/L claim 2 , about 0.6 g/L claim 2 , about 0.7 g/L claim 2 , about 0.8 g/L claim 2 , about 0.9 g/L claim 2 , about 1 g/L claim 2 , about 1.5 g/L claim 2 , about 2 g/L claim 2 , about 2.5 g/L claim 2 , about 3 g/L claim 2 , about 3.5 g/L claim 2 , about 4 g/L claim 2 , about 4.5 g/L claim 2 , about 5 g/L claim 2 , about 5.5 g/L claim 2 , about 6 g/L claim 2 , about 6.5 g/L claim 2 , about 7 g/L claim 2 , about 7.5 g/L claim 2 , about 8 g/L claim 2 , about 8.5 g/L claim 2 , about 9 g/L claim 2 , about 9.5 g/L claim 2 , about 10 g/L claim 2 , about 10.5 g/L claim 2 , about 11 g/L claim 2 , about 12 g/L claim 2 , about 0.2 g/L to about 0.5 g/L claim 2 , about 0.2 g/L to about 1 g/L claim 2 , about 0.2 to about 2 g/L claim 2 , about 0.3 g/L to about 0.6 g/L claim 2 , ...

LARGE SCALE ENZYMATIC SYNTHESIS OF OLIGOSACCHARIDES

A novel UDP-Gal regeneration process and its combined use with a galactosyltransferase to add galactose to a suitable acceptor substrate. Also described herein are synthetic methods for generating Globo-series oligosaccharides in large scale, wherein the methods may involve the combination of a glycosyltransferase reaction and a nucleotide sugar regeneration process. 1. A method for enzymatically synthesizing an oligosaccharide , comprising:(i) producing UDP-Gal from galactose in the presence of a set of UDP-Gal regeneration enzymes, wherein the set of UDP-Gal regeneration enzymes comprises a galactokinase, an UDP-sugar pyrophosphorylase, and a pyruvate kinase; and{'sup': 1A', '1A', '1A, '(ii) converting Lac-ORinto Gb3-ORin the presence of the UDP-Gal and an alpha-1,4 galactosyltransferase, wherein Ris hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or an oxygen protecting group.'}2. The method of claim 1 , wherein the set of UDP-Gal regeneration enzymes further comprises pyrophosphatase.3. The method of claim 1 , wherein (i) and (ii) occur in a Gb3-synthesis reaction mixture comprising galactose claim 1 , PEP claim 1 , ATP claim 1 , UTP claim 1 , the Lac-OR claim 1 , the alpha-1 claim 1 ,4-galactosyltransferase claim 1 , and the set of UDP-Gal regeneration enzymes.4. The method of claim 3 , wherein claim 3 , before occurrence of any enzymatic reactions claim 3 , the molar ratio of the Lac-ORand galactose in the Gb3-synthesis reaction mixture is 1:1.5. The method of claim 1 , wherein Ris hydrogen claim 1 , allyl claim 1 , substituted alkyl claim 1 , biotin claim 1 , or a ceramide.6N. meningitidesE. coliA. thalianaE. coliE. coli.. The method of claim 1 , wherein the alpha-1 claim 1 ,4 galactosyltransferase is LgtC from claim 1 , the ...

ENHANCED CELLODEXTRIN METABOLISM

The present disclosure relates to host cells containing two or more of a recombinant cellodextrin transporter, a recombinant cellodextrin phosphorylase, a recombinant β-glucosidase, a recombinant phosphoglucomutase, or a recombinant hexokinase; and to methods of using such cells for degrading cellodextrin, for producing hydrocarbons or hydrocarbon derivatives from cellodextrin, and for reducing ATP consumption during glucose utilization. 1. A method for degrading cellodextrin , comprising:a) providing a host cell comprising a recombinant cellodextrin transporter and a recombinant polypeptide comprising Y-x(2)-G-x-[KR]-E-N-[AG]-[AG]-[IV]-F-x(2)-[ANST]-[NST]-x(2)-[AIV]-x(2)-[AGT]-x(4)-[AG]-x(4)-[ADNS] (SEQ ID NO: 233), Y-Q-[CN]-M-[IV]-T-F-[CN]-[FILMV]-[AS]-RS-[ST]-[AS]-S-[FY]-[FY]-E-[STV]-G-x-[GS]-R-G-[IM]-G-F-R-D-S-[ACNS]-Q-D-[ILV]-[ILMV]-G-x-V-H-x-[IV]-P-[ADEST]-x-[AV]-[KR]-[AEQ]-x-[IL]-[FIL]-D (SEQ ID NO: 14), or G-x(2)-[FY]-x-N-[AGS]-x-[AS]-W-[APS]-V-[IL]-[AS]-x(2)-A-x(2)-[DE]-x-[AI]-x(3)-[LMV]-[DEN]-[ASV]-[ILV]-x(3)-L-x-T-x(2)-G-[ILV]-x(2)-[SV]-x-P-[AG] (SEQ ID NO: 15), wherein the recombinant polypeptide has cellodextrin phosphorylase activity; andb) culturing the host cell in a medium comprising cellodextrin or a source of cellodextrin, whereby cellodextrin is transported into the cell and degraded by said recombinant polypeptide.2. (canceled)3. (canceled)4. The method of claim 1 , wherein the recombinant polypeptide comprises an amino acid sequence that has at least 29% claim 1 , at least 30% claim 1 , at least 35% claim 1 , at least 40% claim 1 , at least 45% claim 1 , at least 50% claim 1 , at least 55% claim 1 , at least 60% claim 1 , at least 65% claim 1 , at least 70% claim 1 , at least 75% claim 1 , at least 80% claim 1 , at least 85% claim 1 , at least 90% claim 1 , at least 95% claim 1 , at least 99% claim 1 , or at least 100% amino acid identity to the amino acid sequence of CDP_Clent claim 1 , CDP_Ctherm claim 1 , or CDP_Acell.5. The method of claim ...

METHOD FOR PRODUCING SIALIC-ACID-CONTAINING SUGAR CHAIN

[Problem to be Solved] 1. A method for producing a sialylated second sugar chain or a derivative thereof , comprisingreacting a first sugar chain or a derivative thereof with CMP-sialic acid in the presence of sialyltransferase and phosphataseto transfer sialic acid to a non-reducing end of the first sugar chain or a derivative thereof.2. The method according to claim 1 , wherein the first sugar chain or a derivative thereof is a triantennary or tetraantennary N-linked complex sugar chain or a derivative thereof.4. The method according to claim 1 , wherein the sialylated second sugar chain or a derivative thereof is a triantennary or tetraantennary N-linked complex sugar chain claim 1 , wherein the sugar chain is a compound having sialic acid at each of all non-reducing ends or a derivative thereof.7. A method for producing a sugar chain sialylated at its non-reducing end or a derivative thereof claim 1 , comprising the following steps:(a) reacting an agalacto biantennary complex sugar chain or a derivative thereof with UDP-GlcNAc in the presence of MGAT4 and MGAT5;(b) reacting the product of the step (a) with UDP-Gal in the presence of β4GalT1; and(c) reacting the product of the step (b) with CMP-sialic acid in the presence of sialyltransferase and phosphatase.8. The method according to claim 1 , wherein the sialyltransferase is α2 claim 1 ,6-sialyltransferase.9. The method according to claim 1 , wherein the sialyltransferase is human-derived sialyltransferase.10. The method according to claim 1 , wherein the sialyltransferase is ST6Gal-I.11. The method according to claim 1 , wherein the CMP-sialic acid is CMP-Neu5Ac.12. The method according to claim 1 , wherein the phosphatase is alkaline phosphatase.13E. coli. The method according to claim 1 , wherein the phosphatase is -derived alkaline phosphatase.14. A compound having sialic acid at each of all non-reducing ends of a tetraantennary N-linked complex sugar chain or a derivative thereof.15. A compound having α-2 ...

Glucosyltransferase enzymes for production of glucan polymers

Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan.

PRODUCTION OF 3-FUCOSYLLACTOSE AND LACTOSE CONVERTING ALPHA-1,3-FUCOSYLTRANSFERASE ENZYMES

Methods for producing 3-fucosyllactose (3-FL) as well as novel fucosyltransferases, more specifically novel lactose binding alpha-1,3-fucosyltransferase polypeptides, and their applications. Furthermore, methods are provided for producing 3-fucosyllactose (3-FL) using the novel lactose binding alpha-1,3-fucosyltransferases. 139.-. (canceled)40. A method of producing α-1 ,3-fucosyllactose , the method comprising:contacting a polypeptide with a mixture comprising GDP-fucose as donor substrate, and lactose as acceptor substrate, under conditions wherein the polypeptide catalyzes the transfer of a fucose residue from the donor substrate to the acceptor substrate, i) an amino acid sequence comprising a conserved GDP-fucose binding domain [Y/W/L/H/F/M]X[T/S/C][E/Q/D/A][K/R] (SEQ ID NO: 33);', 'ii) an amino acid sequence comprising a conserved [K/D][L/K/M]XXX[F/Y] domain (SEQ ID NO: 34), and', 'iii) if ii) is DM[A/S]VSF (SEQ ID NO: 36), then a conserved motif [N/H]XDPAXLD (SEQ ID NO: 35) is present at the N-terminal region;, 'wherein the polypeptide has α-1,3-fucosyltransferase activity and is able to use lactose as acceptor substrate, wherein the polypeptide compriseswherein X can be any distinct amino acid; andwherein the C-terminus of the polypeptide has less than or equal to 100 amino acids starting from the first amino acid of the GDP-fucose binding domain;so as to thereby produce α-1,3-fucosyllactose.41. The method according to claim 40 , wherein the polypeptide is provided in a cell-free system.42. The method according to claim 40 , wherein the polypeptide is produced by a cell comprising a polynucleotide encoding the polypeptide.43. The method according to claim 42 , wherein GDP-fucose and/or lactose is provided by a cell producing the GDP-fucose and/or lactose.44. The method according to claim 42 , wherein the cell is genetically modified to produce α-1 claim 42 ,3-fucosyllactose claim 42 , and wherein the cell comprises at least one polynucleotide encoding an ...

POLYPEPTIDE

The present invention relates to a polypeptide, or a fragment thereof, capable of enhancing callose biosynthesis and/or accumulation, wherein at least one of the conserved amino acid residues selected from the group consisting of residue corresponding to R84 of SEQ ID NO: 1, residue corresponding to R1926 or SEQ ID NO: 1 and residue corresponding to P189 of SEQ ID NO: 1, of the polypeptide or a fragment thereof, is modified by a mutation selected from the group consisting of substitution and deletion. 1. A polypeptide or a fragment thereof capable of enhancing callose biosynthesis and/or accumulation , characterised in that at least one conserved amino acid residue selected from the group consisting of residue corresponding to R84 of SEQ ID NO: 1 , residue corresponding to R1926 or SEQ ID NO: 1 and residue corresponding to P189 of SEQ ID NO: 1 , of the polypeptide or a fragment thereof is modified by a mutation selected from the group consisting of substitution and deletion.2. The polypeptide according to or a fragment thereof claim 1 , wherein the mutation is a substitution selected from the group consisting of the substitution of R to K and the substitution of P to L.3. The polypeptide according to comprising a sequence having at least 95% claim 1 , preferably at least 99% claim 1 , sequence identity to a sequence selected from the group consisting of SEQ ID NO:s 1-48 claim 1 , or a fragment thereof.4. The polypeptide according to claim comprising a sequence having at least 95% claim 1 , preferably at least 99% claim 1 , sequence identity to a sequence selected from the group consisting of SEQ ID NO:s 1-5 claim 1 , 8 claim 1 , 10 claim 1 , 12-31 claim 1 , 33-40 claim 1 , 42 and 44-47 claim 1 , or a fragment thereof.5. The polypeptide according to comprising a sequence that is at least 95% claim 1 , or at least 99% claim 1 , identical to a sequence selected from the group SEQ ID NO:s 49-55.6. The polypeptide according to or a fragment thereof comprising a sequence ...

PROCESS FOR PRODUCING APLHA-1,3-GLUCAN POLYMER WITH REDUCED MOLECULAR WEIGHT

A process for producing poly alpha-1,3-glucan with reduced molecular weight is disclosed. The process comprises contacting water, sucrose, a polar organic solvent, and a glucosyltransferase enzyme in a solution to produce poly alpha-1,3-glucan. This contacting step results in the production of poly alpha-1,3-glucan having a reduced molecular weight compared to the molecular weight of a poly alpha-1,3-glucan made in the absence of the polar organic solvent. 115-. (canceled)16. A truncated version of SEQ ID NO:32 comprising an amino acid sequence that is at least 90% identical with SEQ ID NO:14 , wherein said truncated version has glucosyltransferase enzyme activity.17. The truncated version of claim 16 , wherein the truncated version comprises an amino acid sequence that is at least 95% identical with SEQ ID NO:14.18. The truncated version of claim 17 , wherein the truncated version comprises an amino acid sequence that is at least 97% identical with SEQ ID NO:14.19. The truncated version of claim 16 , wherein the truncated version consists of an amino acid sequence that is at least 90% identical with SEQ ID NO:14.20. The truncated version of claim 19 , wherein the truncated version consists of an amino acid sequence that is at least 95% identical with SEQ ID NO:14.21. The truncated version of claim 20 , wherein the truncated version consists of an amino acid sequence that is at least 97% identical with SEQ ID NO:14.22. The truncated version of claim 16 , wherein said glucosyltransferase enzyme activity produces poly alpha-1 claim 16 ,3-glucan having at least 50% alpha-1 claim 16 ,3 glycosidic linkages.23. A reaction solution comprising water claim 16 , sucrose and a truncated version of SEQ ID NO:32 comprising an amino acid sequence that is at least 90% identical with SEQ ID NO:14 claim 16 , wherein said truncated version has glucosyltransferase enzyme activity.24. The reaction solution of claim 23 , wherein the truncated version comprises an amino acid sequence ...

GLUCOSYLTRANSFERASE AMINO ACID MOTIFS FOR ENZYMATIC PRODUCTION OF LINEAR POLY ALPHA-1,3-GLUCAN

Reactions comprising water, sucrose, and one or more glucosyltransferase enzymes are disclosed herein. Glucosyltransferase enzymes used in these reactions comprise certain motifs allowing production of insoluble poly alpha-1,3-glucan having at least 95% alpha-1,3 glycosidic linkages. 1. A reaction solution comprising water , sucrose , and a glucosyltransferase enzyme , wherein said glucosyltransferase enzyme comprises a catalytic domain comprising the following three motifs:(i) a motif comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:78,(ii) a motif comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:79, and(iii) a motif comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:80;wherein said glucosyltransferase enzyme does not comprise residues 54-957 of SEQ ID NO:65, residues 55-960 of SEQ ID NO:30, residues 55-960 of SEQ ID NO:4, residues 55-960 of SEQ ID NO:28, or residues 55-960 of SEQ ID NO:20;{'sub': 'w', 'and wherein the glucosyltransferase enzyme produces insoluble poly alpha-1,3-glucan having at least 95% alpha-1,3 glycosidic linkages and a weight average degree of polymerization (DP) of at least 100.'}2. The reaction solution of claim 1 , wherein the catalytic domain comprises an amino acid sequence that is at least 90% identical to amino acid positions 54-957 of SEQ ID NO:65.3. The reaction solution of claim 2 , wherein:(A) the position of the amino acid sequence that is at least 90% identical to SEQ ID NO:78 aligns with amino acid positions 231-243 of SEQ ID NO:65;(B) the position of the amino acid sequence that is at least 90% identical to SEQ ID NO:79 aligns with amino acid positions 396-425 of SEQ ID NO:65; and/or(C) the position of the amino acid sequence that is at least 90% identical to SEQ ID NO:80 aligns with amino acid positions 549-567 of SEQ ID NO:65.4. The reaction solution of claim 1 , wherein motif (i) comprises SEQ ID NO:78 claim 1 , motif (ii) comprises SEQ ID NO ...

Modified glucosyltransferases for producing branched alpha-glucan polymers

Glucosyltransferase enzymes are disclosed herein that produce branched alpha-glucan polymer. Also disclosed, for example, are polynucleotides encoding these enzymes, as well as methods of producing branched alpha-glucan polymer.

Fusion Enzymes

The present disclosure relates to recombinant proteins having N-acetylglucosaminyltransferase activity. The present disclosure further relates to methods for producing complex N-glycans including the steps of providing host cells containing such recombinant proteins and culturing the host cells such that the recombinant proteins are expressed. 169-. (canceled)70. A fungal host cell comprising an expression vector comprising a polynucleotide encoding a fusion protein comprising an N-acetylglucosaminyltransferase I catalytic domain and an N-acetylglucosaminyltransferase II catalytic domain , wherein the N-acetylglucosaminyltransferase II catalytic domain is positioned N-terminal to the N-acetylglucosaminyltransferase I catalytic domain , and wherein the fusion protein catalyzes the transfer of N-acetylglucosamine to a terminal Manα3 residue and N-acetylglucosamine to a terminal Manα6 residue of an acceptor glycan , wherein the acceptor glycan is attached to a heterologous polypeptide.711Trichoderma, Aspergillus, Fusarium, Chrysosporium, Magnaporthe, Mycellopthora, Neurospora,Penicillium.. The fungal host cell of claim , wherein the host cell is selected from the group consisting of or This application claims the benefit of U.S. Provisional Application No. 61/417,144, filed Nov. 24, 2010, which is hereby incorporated by reference in its entirety.The content of the following submission on ASCII text file is incorporated herein by reference in its entirety: a computer readable form (CRF) of the Sequence Listing (file name: 619672001040SEQLIST.txt, date recorded: Nov. 22, 2011, size: 305 KB).The present disclosure relates to compositions and methods useful for the production of N-glycans.Posttranslational modification of proteins is often necessary for proper protein folding and function. A common protein modification is the addition of oligosaccharides (glycans) to nascent polypeptides in the endoplasmic reticulum to form glycoproteins, a process known as glycosylation. ...

Pichia pastoris surface display system

This disclosure relates to novel Pichia pastoris display systems, e.g., display systems featuring the Pichia pastoris strains (such as SuperMan5) with substantially homogeneous N-glycans displayed on cell surface proteins.

ANTIGEN BINDING MOLECULES WITH INCREASED FC RECEPTOR BINDING AFFINITY AND EFFECTOR FUNCTION

The present invention relates to antigen binding molecules (ABMs). In particular embodiments, the present invention relates to recombinant monoclonal antibodies, including chimeric, primatized or humanized antibodies specific for human CD20. In addition, the present invention relates to nucleic acid molecules encoding such ABMs, and vectors and host cells comprising such nucleic acid molecules. The invention further relates to methods for producing the ABMs of the invention, and to methods of using these ABMs in treatment of disease. In addition, the present invention relates to ABMs with modified glycosylation having improved therapeutic properties, including antibodies with increased Fc receptor binding and increased effector function. 1: An isolated polynucleotide comprisinga. a sequence selected from the group consisting of: SEQ ID NO.:5, SEQ ID NO.: 6 and SEQ ID NO:7; andb. a sequence selected from the group consisting of: SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO:23; andc. sequence ID NO:24.2259-. (canceled) This application is a divisional of U.S. patent application Ser. No. 10/981,738, filed Nov. 5, 2004, which claims the benefit of U.S. Provisional Application No. 60/517,096, filed Nov. 5, 2003, the disclosures of which are herein incorporated by reference in their entirety.The content of the following submission on ASCII text file is incorporated herein by reference in its entirety: a computer readable form (CRF) of the Sequence Listing (file name: 146392023111SeqList.txt, date recorded: Oct. 1, 2015, size: 58 KB).The present invention relates to antigen binding molecules (ABMs). In particular embodiments, the present invention relates to recombinant monoclonal antibodies, including chimeric, primatized or humanized antibodies specific for human CD20. In addition, the present invention relates to nucleic acid molecules encoding such ABMs, and vectors and host cells comprising such nucleic acid molecules. The invention further relates to methods for ...

HETERO-TRANSGLYCOSYLASE AND USES THEREOF

The present invention relates to a hetero-transglycosylase protein having cellulose:xyloglucan endotransglucosylase (CXE) activity in addition to mixed-linkage beta-glucan:xyloglucan endotransglucosylase (MXE) activity. The protein may comprise the amino acid sequence of any one of SEQ ID NOs: 2, 6 and 8 or a functional fragment thereof; or an amino acid sequence having at least 60% sequence identity to any one of SEQ ID NO: 2, 6 and 8, or to SEQ ID NO: 2 from amino acid 22 to 280, to SEQ ID NO: 6 from amino acid 26 to 283, or to SEQ ID NO: 8 from amino acid 29 to 287. The invention furthermore relates to an isolated nucleic acid encoding the protein described herein, a chimeric gene comprising, inter alia, the nucleic acid described herein, a vector comprising said chimeric gene, a host cell comprising said vector or said chimeric gene and an according transgenic plant. Further disclosed herein in are a method of producing a transgenic plant and a method of improving properties of cellulosic material. 1. A protein having cellulose:xyloglucan endotransglucosylase activity.2Equisetum.. The protein of which is derived from3. The protein of or comprising(a) the amino acid sequence of any one of SEQ ID NOs: 2, 6 and 8 or a functional fragment thereof; or(b) an amino acid sequence having at least 60% sequence identity to the sequence of any one of SEQ ID NOs: 2, 6 and 8, or(c) an amino acid sequence having at least 60% sequence identity to the sequence of SEQ ID NO: 2 from amino acid 22 to 280, or to the sequence of SEQ ID NO: 6 from amino acid 26 to 283, or to the sequence of SEQ ID NO: 8 from amino acid 29 to 287.4. The protein of any one of to claim 1 , wherein said cellulose:xyloglucan endotransglucosylase activity is one of the predominant activities of the protein.5. An isolated nucleic acid selected from the group consisting of:{'claim-ref': [{'@idref': 'CLM-00001', 'claims 1'}, {'@idref': 'CLM-00004', '4'}], '(a) a nucleic acid sequence encoding the protein of ...

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan. 1. A reaction solution comprising water , sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1 ,3-glucan , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:14.2. The reaction solution of claim 1 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 1 ,3-glucan having at least 50% alpha-1 claim 1 ,3 glycosidic linkages and a number average degree of polymerization of at least 100.3. The reaction solution of claim 2 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 2 ,3-glucan having 100% alpha-1 claim 2 ,3 glycosidic linkages and a number average degree of polymerization of at least 100.4. The reaction solution of claim 3 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 3 ,3-glucan having 100% alpha-1 claim 3 ,3 glycosidic linkages and a number average degree of polymerization of at least 250.5. The reaction solution of claim 1 , further comprising a primer.6. The reaction solution of claim 5 , wherein the primer is dextran.7. The reaction solution of claim 5 , wherein the primer is hydrolyzed glucan.8. A method for producing poly alpha-1 claim 5 ,3-glucan comprising: 'whereby poly alpha-1,3-glucan is produced; and', 'a) contacting at least water, sucrose, and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan, wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:14;'}b) optionally, isolating the poly alpha- ...

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan. 1. A reaction solution comprising water , sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1 ,3-glucan , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:20.2. The reaction solution of claim 1 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 1 ,3-glucan having at least 50% alpha-1 claim 1 ,3 glycosidic linkages and a number average degree of polymerization of at least 100.3. The reaction solution of claim 2 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 2 ,3-glucan having 100% alpha-1 claim 2 ,3 glycosidic linkages and a number average degree of polymerization of at least 100.4. The reaction solution of claim 3 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 3 ,3-glucan having 100% alpha-1 claim 3 ,3 glycosidic linkages and a number average degree of polymerization of at least 250.5. The reaction solution of claim 1 , further comprising a primer.6. The reaction solution of claim 5 , wherein the primer is dextran.7. The reaction solution of claim 5 , wherein the primer is hydrolyzed glucan.8. A method for producing poly alpha-1 claim 5 ,3-glucan comprising: 'whereby poly alpha-1,3-glucan is produced; and', 'a) contacting at least water, sucrose, and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan, wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:20;'}b) optionally, isolating the poly alpha- ...

Glucosyltransferase enzymes for production of glucan polymers

Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan.

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan. 1. A reaction solution comprising water , sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1 ,3-glucan , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:28.2. The reaction solution of claim 1 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 1 ,3-glucan having at least 50% alpha-1 claim 1 ,3 glycosidic linkages and a number average degree of polymerization of at least 100.3. The reaction solution of claim 2 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 2 ,3-glucan having 100% alpha-1 claim 2 ,3 glycosidic linkages and a number average degree of polymerization of at least 100.4. The reaction solution of claim 3 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 3 ,3-glucan having 100% alpha-1 claim 3 ,3 glycosidic linkages and a number average degree of polymerization of at least 250.5. The reaction solution of claim 1 , further comprising a primer.6. The reaction solution of claim 5 , wherein the primer is dextran.7. The reaction solution of claim 5 , wherein the primer is hydrolyzed glucan.8. A method for producing poly alpha-1 claim 5 ,3-glucan comprising: 'whereby poly alpha-1,3-glucan is produced; and', 'a) contacting at least water, sucrose, and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan, wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:28;'}b) optionally, isolating the poly alpha- ...

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan. 1. A reaction solution comprising water , sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1 ,3-glucan , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:30.2. The reaction solution of claim 1 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 1 ,3-glucan having at least 50% alpha-1 claim 1 ,3 glycosidic linkages and a number average degree of polymerization of at least 100.3. The reaction solution of claim 2 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 2 ,3-glucan having 100% alpha-1 claim 2 ,3 glycosidic linkages and a number average degree of polymerization of at least 100.4. The reaction solution of claim 3 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 3 ,3-glucan having 100% alpha-1 claim 3 ,3 glycosidic linkages and a number average degree of polymerization of at least 250.5. The reaction solution of claim 1 , further comprising a primer.6. The reaction solution of claim 5 , wherein the primer is dextran.7. The reaction solution of claim 5 , wherein the primer is hydrolyzed glucan.8. A method for producing poly alpha-1 claim 5 ,3-glucan comprising: 'whereby poly alpha-1,3-glucan is produced; and', 'a) contacting at least water, sucrose, and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan, wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:30;'}b) optionally, isolating the poly alpha- ...

Heat stable mutants of starch biosynthesis enzymes

The subject invention pertains to novel mutant polynucleotide molecules that encode enzymes that have increased heat stability. These polynucleotides, when expressed in plants, result in increased yield in plants grown under conditions of heat stress. The polynucleotide molecules of the subject invention encode maize endosperm ADP glucose pyrophosphorylase (AGP) and soluble starch synthase (SSS) enzyme activities. Plants and plant tissue bred to contain, or transformed with, the mutant polynucleotides, and expressing the polypeptides encoded by the polynucleotides, are also contemplated by the present invention. The subject invention also concerns methods for isolating polynucleotides and polypeptides contemplated within the scope of the invention. Methods for increasing yield in plants grown under conditions of heat stress are also provided.

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan. 1. A reaction solution comprising water , sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1 ,3-glucan , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:10.2. The reaction solution of claim 1 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 1 ,3-glucan having at least 50% alpha-1 claim 1 ,3 glycosidic linkages and a number average degree of polymerization of at least 100.3. The reaction solution of claim 2 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 2 ,3-glucan having 100% alpha-1 claim 2 ,3 glycosidic linkages and a number average degree of polymerization of at least 100.4. The reaction solution of claim 3 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1 claim 3 ,3-glucan having 100% alpha-1 claim 3 ,3 glycosidic linkages and a number average degree of polymerization of at least 250.5. The reaction solution of claim 1 , further comprising a primer.6. The reaction solution of claim 5 , wherein the primer is dextran.7. The reaction solution of claim 5 , wherein the primer is hydrolyzed glucan.8. A method for producing poly alpha-1 claim 5 ,3-glucan comprising: 'whereby poly alpha-1,3-glucan is produced; and', 'a) contacting at least water, sucrose, and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan, wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:10;'}b) optionally, isolating the poly alpha- ...

Glucosyltransferase enzymes for production of glucan polymers

Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan.

GLYCOSYLATION MODIFICATION OF BIOACTIVE COMPOUNDS AND DRUGS BY PLANT GLYCOSYLTRANSFERASES (UGTS)

In alternative embodiments, provided are methods for the glycosylation modification of bioactive compounds and drugs using isolated, recombinant or genetically modified uridine diphosphate glycosyl-transferases (UGTs). In alternative embodiments, provided are methods for modifying UGTs to generate recombinant UGTs with altered donor and/or acceptor specificities. In alternative embodiments, provided are methods for screening for recombinantly engineered UGTs with new or altered properties, for example, for new or altered donor and/or acceptor specificities, where in alternative embodiments the screening comprise use of bacterial, yeast or baculovirus expression system. 1. A method for identifying or screening for a recombinant or genetically modified uridine diphosphate glycosyl-transferase (UGT) having a modified sequence such that the modification of the UGT results in the glycosylation of or adding a sugar moiety to an otherwise unglycosylated bioactive compound or target drug , or results in generating a modified glycosylation of a bioactive compound or a target drug by adding a sugar moiety , the method comprising:(a) providing or having provided a recombinant or genetically modified UGT, wherein the expressed recombinant or genetically modified UGT has an altered or new donor and/or acceptor specificity,(b) providing or having provided an acceptor molecule,(c) expressing the recombinant or genetically modified UGT in an expression system,(d) contacting the recombinant or genetically modified UGT with a bioactive compound or a target drug in the expression system, and(e) screening for the generation of a UGT that results in the glycosylation of an otherwise unglycosylated acceptor molecule, or results in generating a modified glycosylation of the acceptor molecule.2. The method of claim 1 , wherein the recombinant or genetically modified UGT uses UDP-glucose claim 1 , UDP-glucuronic acid and/or UDP-rhamnose as a donor claim 1 , thereby adding a glucose claim 1 ...

OVEREXPRESSION OF N-GLYCOSYLATION PATHWAY REGULATORS TO MODULATE GLYCOSYLATION OF RECOMBINANT PROTEINS

Methods of modulating the properties of a cell culture expressing a protein of interest are provided. In various embodiments the methods relate to the overexpression of proteins involved in the N-glycosylation pathway. 110.-. (canceled)11. A method of regulating the high mannose glycoform content of a recombinant protein during a mammalian cell culture process , comprising:transfecting a mammalian host cell to overexpress a protein that is involved in an N-glycosylation pathway, wherein the protein involved in the N-glycosylation pathway is selected from the group consisting of N-acetyl-glucosaminyltransferase-1 (encoded by Mgat1); N-acetyl-glucosaminyltransferase-2 (encoded by Mgat2); a UDP-Galactose transporter encoded by Slc35a2; and combinations thereof, wherein the regulated high mannose glycoform content of the recombinant protein is less than or equal to 10%; andwherein the mammalian cell culture is perfused using alternating tangential flow (ATF).12. The method of claim 11 , wherein perfusion begins on or about day 1 to on or about day 9 of the cell culture.13. The method of claim 11 , wherein perfusion begins on or about day 3 to on or about day 7 of the cell culture.14. The method of claim 11 , wherein perfusion begins when the cells have reached a production phase.15. The method of claim 14 , wherein perfusion is accomplished by alternating tangential flow using an ultrafilter or a microfilter.16. A method of regulating the high mannose glycoform content of a recombinant protein during a mammalian cell culture process claim 14 , comprising:transfecting a mammalian host cell to overexpress a protein that is involved in an N-glycosylation pathway, wherein the protein involved in the N-glycosylation pathway is selected from the group consisting of N-acetyl-glucosaminyltransferase-1 (encoded by Mgat1); N-acetyl-glucosaminyltransferase-2 (encoded by Mgat2); a UDP-Galactose transporter encoded by Slc35a2; and combinations thereof, wherein the regulated high ...

METABOLICALLY ENGINEERED ORGANISMS FOR THE PRODUCTION OF ADDED VALUE BIO-PRODUCTS

The present invention relates to genetically engineered organisms, especially microorganisms such as bacteria and yeasts, for the production of added value bio-products such as specialty saccharide, activated saccharide, nucleoside, glycoside, glycolipid or glycoprotein. More specifically, the present invention relates to host cells that are metabolically engineered so that they can produce said valuable specialty products in large quantities and at a high rate by bypassing classical technical problems that occur in biocatalytical or fermentative production processes. 1. A metabolically engineered organism for the production of at least one specialty product chosen from the group consisting of a saccharide , an activated saccharide , a nucleoside , a glycoside , a glycolipid and a glycoprotein , characterized in that:a) said organism is genetically modified with the introduction of at least: i) a gene encoding for a carbohydrate hydrolase in combination with a gene encoding for a carbohydrate kinase, ii) a gene encoding for a carbohydrate synthase, or, iii) a gene encoding for a carbohydrate phosphorylase, so that said organism is capable to split a disaccharide, oligosaccharide, polysaccharide or a mixture thereof into an activated saccharide and a saccharide, andb) said organism is further genetically modified so that at least one other gene than any of the introduced genes of step a) of said organism is rendered less-functional or non-functional and wherein said other gene encodes for an enzyme which converts said activated saccharide into biomass and/or bio-catalytic enzymes.2. A metabolically engineered organism according to wherein said organism is further genetically modified with the introduction of at least one other gene which converts said activated saccharide into a specialty product claim 1 , or claim 1 , wherein at least one other endogenous gene of said organism which converts said activated saccharide into a specialty product is over-expressed.3. A ...

CATALYTIC DOMAINS OF BETA(1,4)-GALACTOSYLTRANSFERASE I HAVING ALTERED METAL ION SPECIFICITY

Disclosed are mutants of galactosyltransferases that can catalyze formation of oligosaccharides in the presence of magnesium; mutants of galactosyltransferases having altered donor and acceptor specificity which can catalyze formation of oligosaccharides in the presence of magnesium; methods and compositions that can be used to synthesize oligosaccharides; methods for increasing the immunogenicity of an antigen; and methods to stabilize platelets. 1. A purified and isolated catalytic domain from a β(1 ,4)-galactosyltransferase I that catalyzes formation of a galactose-β(1 ,4)-N-acetylglucosamine bond in the presence of magnesium.2. The catalytic domain according to claim 1 , wherein the rate of formation of the galactose-β(1 claim 1 ,4)-N-acetylglucosamine bond is at least two-fold claim 1 , five-fold claim 1 , ten-fold claim 1 , or one hundred-fold greater than wild-type β(1 claim 1 ,4)-galactosyltransferase in the presence of magnesium.3. The catalytic domain according to claim 1 , wherein the catalytic domain has a conservative amino acid exchange at an amino acid position corresponding to amino acid position 344 of SEQ ID NO: 6.4. The catalytic domain according to claim 3 , wherein histidine is exchanged for methionine at an amino acid position corresponding to amino acid position 344 of SEQ ID NO: 6.5. The catalytic domain according to claim 1 , further comprising a conservative amino acid substitution at an amino acid position corresponding to amino acid position 342 of SEQ ID NO: 6.6. The catalytic domain according to claim 5 , wherein threonine is exchanged for cysteine at amino acid position 342.7. A polypeptide comprising the catalytic domain according to .8. A purified and isolated catalytic domain from a β(1 claim 1 ,4)-galactosyltransferase I that catalyzes formation of a glucose-β(1 claim 1 ,4)-N-acetylglucosamine bond in the presence of magnesium.9. The catalytic domain according to claim 8 , wherein the rate of formation of the glucose-β(1 claim 8 ,4 ...

METABOLICALLY ENGINEERED ORGANISMS FOR THE PRODUCTION OF ADDED VALUE BIO-PRODUCTS

The present invention relates to genetically engineered organisms, especially microorganisms such as bacteria and yeasts, for the production of added value bio-products such as specialty saccharide, activated saccharide, nucleoside, glycoside, glycolipid or glycoprotein. More specifically, the present invention relates to host cells that are metabolically engineered so that they can produce said valuable specialty products in large quantities and at a high rate by bypassing classical technical problems that occur in biocatalytical or fermentative production processes. 1. A metabolically engineered bacterium or yeast for production of a carbohydrate specialty product , characterized in that said bacterium or yeast:a) has been genetically modified by introducing a heterologous gene encoding a sucrose phosphorylase capable of splitting sucrose into glucose-1-phosphate and fructose, or a heterologous gene encoding a carbohydrate hydrolase capable of splitting sucrose into glucose and fructose;b) comprises a fructokinase to catalyze conversion of fructose to fructose-6-phosphate; andc) has been further genetically modified to prevent loss of fructose-6-phosphate via glycolysis due to genetic disruption of an endogenous gene encoding a phosphofructokinase, a phosphoglucose isomerase, or a combination thereof.2. The metabolically engineered bacterium or yeast according to claim 1 , wherein an endogenous gene encoding a phosphofructokinase has been disrupted.3. The metabolically engineered bacterium or yeast according to claim 1 , wherein an endogenous gene encoding a phosphoglucose isomerase has been disrupted.4. The metabolically engineered bacterium or yeast according to claim 1 , wherein an endogenous gene encoding a phosphofructokinase and an endogenous gene encoding a phosphoglucose isomerase have been disrupted.5. The metabolically engineered bacterium or yeast according to claim 1 , wherein said bacterium or yeast has been further genetically modified to prevent ...

HIGH MOLECULAR WEIGHT HEPAROSAN POLYMERS AND METHODS OF PRODUCTION AND USE THEREOF

High molecular weight heparosan polymers are described, as are methods of producing and using the high molecular weight heparosan polymers. 1. A method for recombinantly producing a high molecular weight heparosan polymer , the method comprising the steps of: (a) the polypeptide having heparosan synthase activity is at least 90% identical to at least one of SEQ ID NOS:2, 4, and 6-8, and the nucleotide sequence encoding the polypeptide has been gene-optimized for expression in the recombinant host cell;', '(b) the polypeptide having heparosan synthase activity has 1-20 amino acid additions, deletions, and/or substitutions when compared to at least one of SEQ ID NOS:2, 4, and 6-8, and the nucleotide sequence encoding the polypeptide has been gene-optimized for expression in the recombinant host cell;', '(c) the polypeptide is encoded by the nucleotide sequence of at least one of SEQ ID NOS:9-11;', '(d) the polypeptide is encoded by a nucleotide sequence that is at least 90% identical to at least one of SEQ ID NOS:9-11; and, 'culturing a recombinant host cell containing a nucleotide sequence encoding a polypeptide having heparosan synthase activity under conditions appropriate for the expression of the heparosan synthase, wherein at least one of{'sub': 'n', 'isolating heparosan polymer produced by the heparosan synthase, wherein the isolated heparosan polymer is biocompatible with a mammalian patient and biologically inert within extracellular compartments of a mammalian patient, and wherein the isolated heparosan polymer is represented by the structure (-GlcUA-beta1,4-GlcNAc-alpha-1,4-), wherein n is a positive integer greater than or equal to 2,000.'}2. The method of claim 1 , wherein the isolated heparosan polymer is further defined as having a value for n in a range of from about 2 claim 1 ,000 to about 17 claim 1 ,000.3. The method of claim 1 , wherein the recombinant host cell further comprises at least one gene encoding an enzyme for synthesis of a heparosan ...

Microorganisms and Methods for Producing Sialylated and N-Acetylglucosamine-Containing Oligosaccharides

The invention provides compositions and methods for engineering bacteria to produce sialylated and N-acetylglucosamine-containing oligosaccharides, and the use thereof in the prevention or treatment of infection. 1. A method for producing a sialylated oligosaccharide in a bacterium comprising:providing a bacterium, said bacterium comprising an exogenous sialyl-transferase, a deficient sialic acid catabolic pathway, a sialic acid synthetic capability, and a functional lactose permease gene; andculturing said bacterium in the presence of lactose.2. The method of claim 1 , wherein said deficient sialic acid catabolic pathway comprises a mutation in any one of the genes selected from endogenous N-acetylneuraminate lyase (nanA) gene claim 1 , endogenous N-acetylmannosamine kinase gene (nanK) claim 1 , endogenous N-acetylmannosamine-6-phosphate epimerase gene (nanE) claim 1 , and endogenous N-acetylneuraminic acid transporter gene (nanT) claim 1 , or any combination thereof.3. The method of claim 1 , wherein said deficient sialic acid catabolic pathway comprises a mutation in endogenous N-acetylneuraminate lyase (nanA) gene claim 1 , and optionally claim 1 , a mutation in endogenous N-acetylneuraminic acid transporter gene (nanT).4. The method of claim 2 , wherein said deficient sialic acid catabolic pathway further comprises an endogenous N-acetylmannosamine kinase gene (nanK) and endogenous N-acetylmannosamine-6-phosphate epimerase gene (nanE) that are not mutated.5. The method of claim 1 , wherein said deficient sialic acid catabolic pathway comprises a mutation in endogenous N-acetylneuraminate lyase (nanA) gene claim 1 , a mutation in endogenous N-acetylmannosamine-6-phosphate epimerase gene (nanE) claim 1 , and optionally claim 1 , a mutation in endogenous N-acetylneuraminic acid transporter gene (nanT).6. The method of claim 1 , wherein the mutation comprises a null mutation.7. The method of claim 5 , wherein said deficient sialic acid catabolic pathway further ...

Microbial production of steviol glycosides

The invention provides methods for making steviol glycosides, including RebM and glycosylation products that are minor products in stevia leaves, and provides enzymes, encoding polynucleotides, and host cells for use in these methods. The invention provides engineered enzymes and engineered host cells for producing steviol glycosylation products, such as RebM, at high purity and/or yield. The invention further provides methods of making products containing steviol glycosides, such as RebM, including food products, beverages, oral care products, sweeteners, and flavoring products.

COMPOSITIONS AND METHODS RELATING TO A MUTANT CLOSTRIDIUM DIFFICILE TOXIN

In one aspect, the invention relates to an immunogenic composition that includes a mutant toxin A and/or a mutant toxin B. The mutant toxin may include a glucosyltransferase domain having at least one mutation and a cysteine protease domain having at least one mutation, relative to the corresponding wild-type toxin. The mutant toxins may include at least one amino acid that is chemically crosslinked. In another aspect, the invention relates to methods and compositions for use in culturing and in producing toxins. 1. An immunogenic composition comprising a lyophilized polypeptide comprising the amino acid sequence set forth in any one of SEQ ID NOs: 178-761; a saponin adjuvant; and a stabilizer selected from the group consisting of sorbitol , mannitol , starch , dextran , sucrose , trehalose , lactose , and glucose; wherein the composition does not comprise formaldehyde.2. The composition according to claim 1 , wherein the lyophilized polypeptide comprises at least 90% purity by weight of the total protein content in the composition.3. The composition according to claim 2 , wherein the lyophilized polypeptide comprises at least 95% purity by weight of the total protein content in the composition.4. The composition according to claim 2 , wherein the lyophilized polypeptide comprises at least 97% purity by weight of the total protein content in the composition.5. The composition according to claim 2 , wherein the purity is measured by size exclusion chromatography.6. The composition according to claim 1 , wherein the stabilizer comprises sucrose.7. The composition according to claim 1 , wherein the stabilizer comprises trehalose.8. The composition according to claim 1 , wherein the saponin adjuvant comprises an immune stimulating Complex (ISCOM).9. The composition according to claim 1 , wherein the saponin adjuvant comprises QS-2110. The composition according to claim 1 , wherein the composition further comprises 3-De-O-acylated monophosphoryl lipid A.11. The ...

FORMULATIONS COMPRISING POLYMERIC XYLOGLUCAN AS A CARRIER FOR AGRICULTURALLY BENEFICIAL AGENTS

The present invention relates to formulations comprising one or more agriculturally beneficial agents formulated with polymeric xyloglucan as a carrier and their use. 1. A formulation comprising one or more agriculturally beneficial agents formulated with a composition selected from the group consisting of (a) a composition comprising a xyloglucan endotransglycosylase , a polymeric xyloglucan , and a functionalized xyloglucan oligomer comprising a chemical group; (b) a composition comprising a xyloglucan endotransglycosylase , a polymeric xyloglucan functionalized with a chemical group , and a functionalized xyloglucan oligomer comprising a chemical group; (c) a composition comprising a xyloglucan endotransglycosylase , a polymeric xyloglucan functionalized with a chemical group , and a xyloglucan oligomer; (d) a composition comprising a xyloglucan endotransglycosylase , a polymeric xyloglucan , and a xyloglucan oligomer; (e) a composition comprising a xyloglucan endotransglycosylase and a polymeric xyloglucan functionalized with a chemical group; (f) a composition comprising a xyloglucan endotransglycosylase and a polymeric xyloglucan; (g) a composition comprising a xyloglucan endotransglycosylase and a functionalized xyloglucan oligomer comprising a chemical group; (h) a composition comprising a xyloglucan endotransglycosylase and a xyloglucan oligomer , and (i) a composition of (a) , (b) , (c) , (d) , (e) , (f) , (g) , or (h) without a xyloglucan endotransglycosylase , wherein the formulation provides an agricultural benefit.2. The formulation of claim 1 , wherein the one or more agriculturally beneficial agents are linked to claim 1 , coated by claim 1 , embedded in claim 1 , or encapsulated by the polymeric xyloglucan or the polymeric xyloglucan functionalized with a chemical group.3. The formulation of claim 2 , wherein the linking of the one or more agriculturally beneficial agents to the polymeric xyloglucan or the polymeric xyloglucan functionalized with a ...

DITERPENE PRODUCTION

The present invention relates to a recombinant microorganism comprising one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity, whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol. The recombinant microorganism may also be capable of expressing one or more UDP-glucosyltransferases such that the microorganism is capable of producing one or more steviol glycosides. 1. A recombinant microorganism comprising at least one nucleotide sequence encoding:a polypeptide having ent-copalyl pyrophosphate synthase activity;a polypeptide having ent-Kaurene synthase activity;a polypeptide having ent-Kaurene oxidase activity; anda polypeptide having kaurenoic acid 13-hydroxylase activity,whereby expression of the nucleotide sequence confers on the microorganism an ability to produce at least steviol.2. A recombinant microorganism according to claim 1 , wherein the microorganism comprises one or more nucleotide sequences encoding a polypeptide having UDP-glucosyltransferase activity claim 1 ,whereby expression of the nucleotide sequence confers on the microorganism an ability to produce at least one of steviolmonoside, steviolbioside, stevioside or rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, rubusoside, dulcoside A.3. A recombinant microorganism according to claim 2 , wherein the microorganism comprises a nucleotide sequence encoding a polypeptide capable of catalyzing an addition of a C-13-glucose to steviol claim 2 ,whereby expression of the nucleotide sequence confers on the microorganism an ability to produce at least steviolmonoside.4. A recombinant microorganism according to claim 2 , wherein the microorganism comprises a nucleotide sequence ...

METHOD FOR IMPROVING CROP PRODUCTIVITY

Plants and plant cells derived therefrom are disclosed having one or more improved or introduced desirable traits such as increased plant biomass including both increased shoot and root growth and increased seed yield. Also disclosed is a method for the production of plants with increased cold tolerance, salt tolerance, drought tolerance, and resistance to parasitic plants. Plants can be produced which have an exogenous polynucleotide molecule encoding a glycosyltransferase (UGT) and/or an exogenous polynucleotide molecule which increases transcription of an endogenous UGT. 1. A plant cell of , or derived from , a plant comprising an exogenous polynucleotide molecule encoding a uridine diphosphate glucose (UDP) glycosyltransferase (UGT) and/or an exogenous polynucleotide molecule which increases transcription of an endogenous gene encoding a UGT , wherein said UGT is a UGT which glycosylates one or more strigolactone or strigolactone-like compound (ie an SL-UGT).2. The plant cell of claim 1 , wherein the plant cell is selected from crop and forage species of plants claim 1 , and hybrids thereof.3Brassica napus. The plant cell of claim 2 , wherein the plant cell is of the species (canola).4. The plant cell of claim 1 , wherein the plant cell is selected from species and hybrids of lawn grasses and ornamental plants.5. The plant cell of claim 1 , which forms part of a multicellular structure.6. The plant cell of claim 1 , wherein the exogenous polynucleotide molecule encodes a SL-UGT.7. The plant cell of claim 6 , wherein the exogenous polynucleotide molecule encodes a SL-UGT comprising an amino acid sequence that has at least 35% sequence identity to the amino acid sequence shown as SEQ ID NO: 3 or a biologically active fragment thereof.8. The plant cell of claim 6 , wherein the exogenous polynucleotide molecule encodes a SL-UGT comprising an amino acid sequence that has at least 98% sequence identity to the amino acid sequence shown as SEQ ID NO: 3 or a biologically ...

CREATION OF CHRYSANTHEMUM WITH BLUE FLOWER COLOR

Provided are transformed plants having blue flower color, their self-fertilized progenies or cross-fertilized progenies thereof, a vegetative propagated plants thereof, and a part, a tissue or a cell of the plant body. Anthocyanin 3′,5′-O-glucosyltransferase gene (CtA3′5′GT) derived from and flavonoid 3′,5′-hydroxylase gene derived from (CamF3′5′H) are coexpressed in petals. 1: An expression cassette comprising: (1-a) a polynucleotide comprising the nucleotide sequence listed as SEQ ID NO: 1;', '(1-b) a polynucleotide that hybridizes with a polynucleotide comprising the nucleotide sequence complementary to the nucleotide sequence listed as SEQ ID NO: 1 under stringent conditions, the polynucleotide encoding a protein with activity of transferring sugars to the and 5′-hydroxyl groups of anthocyanins;', '(1-c) a polynucleotide encoding a protein comprising the amino acid sequence listed as SEQ ID NO: 2;', '(1-d) a polynucleotide comprising an amino acid sequence which is the amino acid sequence listed as SEQ ID NO: 2 with a deletion, substitution, insertion and/or addition of one or more amino acids, and encoding a protein with activity of transferring sugars to the 3′- and 5′-hydroxyl groups of anthocyanins; and', '(1-e) a polynucleotide having an amino acid sequence with at least 90% identity with the amino acid sequence listed as SEQ ID NO: 2, and encoding a protein with activity of transferring sugars to the 3′- and 5′-hydroxyl groups of anthocyanins, and, 'a first polynucleotide selected from the group consisting of the following (1-a) to (1-e) (2-a) a polynucleotide comprising the nucleotide sequence listed as SEQ ID NO: 3;', '(2-b) a polynucleotide that hybridizes with a polynucleotide comprising the nucleotide sequence complementary to the nucleotide sequence listed as SEQ ID NO: 3 under stringent conditions, the polynucleotide encoding a protein with activity of hydroxylating the 3′- and 5′-positions of flavonoids;', '(2-c) a polynucleotide encoding a protein ...

FERMENTATION PROCESS FOR PRODUCING MONOSACCHARIDES IN FREE FORM FROM NUCLEOTIDE-ACTIVATED SUGARS

The present invention relates to a process for producing a monosaccharide, e.g. L-fucose, in free form using a microbial fermentation process. The used microorganism exhibits hydrolase activity on nucleotide-activated sugars and releases the monosaccharide in an unmodified free form. The free monosaccharide is retrieved from the supernatant of the cultivated microorganism. 1. Process for producing a monosaccharide of interest in free form using a microorganism , the process comprisinga.) providing a microorganism for the synthesis of the monosaccharide comprising an enzyme capable of catalyzing the hydrolysis of a nucleotide-activated monosaccharide to release the monosaccharide of interest from the nucleotide-activated monosaccharide, andb.) cultivating the microorganism in a medium suitable for growing the microorganism, wherein the microorganism is unable to metabolize the monosaccharide to a significant extent, so that the monosaccharide of interest accumulates during cultivation.2. The process of claim 1 , wherein a recombinant microorganism is used claim 1 , wherein the recombinant microorganism comprises a heterologous nucleic acid encoding an enzyme capable of catalyzing the hydrolysis of a nucleotide-activated monosaccharide.3. The process of claim 1 , wherein the enzyme is a glycosyltransferase claim 1 , optionally a fucosyltransferase claim 1 , the enzyme being able to catalyze the hydrolysis of the nucleotide-activated monosaccharide GDP-fucose in the absence of an acceptor molecule.4Escherichia coliHelicobacter pylori. The process of claim 1 , wherein the enzyme is a variant of the 2-fucosyltransferase encoded by the wbgL gene from claim 1 , or a variant of the 1 claim 1 ,2-fucosyltransferase encoded by the futC gene from claim 1 , the variant carrying at least one claim 1 , optionally at least two claim 1 , and optionally more than two modifications as compared to the wild type 2-fucosyltransferase encoded by the wbgL gene or to the wild type 1 claim 1 ...

Microbial production of triterpenoids including mogrosides

The present invention provides host cells and methods for making mogrol glycosides, including Mogroside V (Mog. V), Mogroside VI (Mog. VI), Iso-Mogroside V (Isomog. V), and glycosylation products that are minor products in Siraitia grosvenorii. The invention provides engineered enzymes and engineered host cells for producing mogrol glycosylation products, such as Mog, V. Mog. VI, and Isomog. V, at high purity and/or yield. The present technology further provides methods of making products containing mogrol glycosides, such as Mog. V, Mog. VI, and Isomog. V, including food products, beverages, oral care products, sweeteners, and flavoring products.

TRANSGENIC SILKWORM HAVING MAMMALIAN-TYPE SUGAR CHAIN ATTACHED THERETO

It is intended to develop and provide a technique of conveniently allowing a transgenic silkworm by itself and at an individual level to produce a recombinant protein having a mammalian-type sugar chain sialic acid attached thereto, without the need of a baculovirus expression system or oral and transdermal administration of sialic acid. An expression vector was developed which can induce the expression of a mammalian-type glycosylation-related gene group only in a silk gland such that the recombinant protein modified with the mammalian-type sugar chain has no adverse effect on the silkworm itself. A transgenic silkworm harboring the expression vector was prepared. 1. A mammalian-type glycosylation agent comprising one to three independent expression vector(s) comprisinga silk-spinning insect-derived middle and/or posterior silk gland promoter anda gene encoding β1,4-galactosyltransferase or a nucleotide encoding an active fragment of the enzyme, functionally linked downstream of the promoter, andgenes encodingUDP-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase,α2,6-sialyltransferase, andNeu5Ac9-phosphate synthase and/or Neu5Ac9-phosphate phosphatase, ornucleotides encoding active fragments of the enzymes, whereinthe genes encoding the enzymes or the nucleotides encoding active fragments of the enzymes are arranged so as to be under direct or indirect expression control of the middle and/or posterior silk gland promoter.2. The mammalian-type glycosylation agent according to claim 1 , wherein the β1 claim 1 ,4-galactosyltransferase is GalT2.3. The mammalian-type glycosylation agent according to claim 1 , wherein the middle silk gland promoter is a promoter of sericin 1 gene claim 1 , sericin 2 gene claim 1 , or sericin 3 gene.4. The mammalian-type glycosylation agent according to claim 1 , wherein the posterior silk gland promoter is a promoter of fibroin H chain gene claim 1 , fibroin L chain gene claim 1 , or p25 gene.5. The mammalian-type glycosylation ...

VACCINES AGAINST CLOSTRIDIUM DIFFICILE COMPRISING RECOMBINANT TOXINS

The present invention relates to recombinant toxin A (TcdA) and toxin B (TcdB) and binary toxin A (CDTa) proteins comprising specifically defined mutations relative to the native toxin sequence that substantially reduce or eliminate toxicity. The invention also relates to vaccines and immunogenic compositions comprising these recombinant toxins, as well as combinations of these toxins with binary toxin B (CDTb), which are capable of providing protection against infection and/or the effects thereof. The invention also relates to methods of inducing an immune response to comprising administering the vaccines and immunogenic compositions described herein to a patient. The invention also encompasses methods of expressing recombinant toxin A and toxin B and CDTa mutants and CDTb in recombinant expression systems. In exemplary embodiments, TcdA, TcdB, and CDTa mutant toxins comprising sufficient mutations to substantially reduce or eliminate toxicity are expressed in the baculovirus/insect cell expression system. 1C. difficile. A recombinant toxin A (TcdA) protein comprising at least 4 mutations selected from the group consisting of: W101A , D287A , E514Q , D285A , S517A , W519A , and C700A.2. The TcdA protein of claim 1 , wherein the protein comprises a set of mutations selected from the group consisting of: (a) W101A claim 1 , D287A claim 1 , E514Q claim 1 , and W519A; (b) W101A claim 1 , D287A claim 1 , E514Q and D285A; (c) W101A claim 1 , D287A claim 1 , E514Q and S517A; (d) W101A claim 1 , D287A claim 1 , and E514Q; and (e) W101A claim 1 , D287A claim 1 , E514Q claim 1 , W519A claim 1 , and C700A.3. The TcdA protein of claim 2 , wherein the protein comprises W101A claim 2 , D287A claim 2 , E514Q claim 2 , W519A claim 2 , and C700A mutations.4C. difficile. The TcdA protein of claim 2 , wherein the TcdA protein comprises a sequence of amino acids that is derived from NAP 1 reference strain or VPI 10463 reference strain.5. The TcdA protein of claim 1 , wherein the ...

ALPHA (1,2) FUCOSYLTRANSFERASE SYNGENES FOR USE IN THE PRODUCTION OF FUCOSYLATED OLIGOSACCHARIDES

The invention provides compositions and methods for engineering or other host production bacterial strains to produce fucosylated oligosaccharides, and the use thereof in the prevention or treatment of infection. 1. A method for producing a fucosylated oligosaccharide in a bacterium comprisingproviding bacterium comprising an exogenous lactose-utilizing α(1,2) fucosyltransferase enzyme, wherein said α(1,2) fucosyltransferase enzyme has at least 90% sequence identity to amino acid sequence SEQ ID NO: 17; andculturing said bacterium in the presence of lactose.2. (canceled)3Prevotellaa. The method of claim 1 , wherein said α(1 claim 1 ,2) fucosyltransferase enzyme comprises sp. FutW claim 1 , or a functional variant or fragment thereof.4. (canceled)5. The method of claim 1 , further comprising retrieving the fucosylated oligosaccharide from said bacterium or from a culture supernatant of said bacterium.6. The method of claim 1 , wherein said fucosylated oligosaccharide comprises 2′-fucosyllactose (2′-FL) claim 1 , lactodifucotetraose (LDFT) claim 1 , or lacto-N-difucohexaose I (LDFH I).7. The method of claim 1 , wherein the bacterium further comprises an exogenous lactose-utilizing α(1 claim 1 ,3) fucosyltransferase enzyme and/or an exogenous lactose-utilizing α(1 claim 1 ,4) fucosyltransferase enzyme claim 1 , or wherein said bacterium further comprises a reduced level of β-galactosidase activity claim 1 , a defective colanic acid synthesis pathway claim 1 , an inactivated adenosine-5′-triphosphate (ATP)-dependent intracellular protease claim 1 , or an inactivated endogenous lacA gene claim 1 , or any combination thereof.8Helicobacter pylori. The method of claim 7 , wherein the exogenous lactose-utilizing α(1 claim 7 ,3) fucosyltransferase enzyme comprises a 26695 futA gene.9Helicobacter pyloriHelicobacter pylori. The method of claim 7 , wherein the exogenous lactose-utilizing α(1 claim 7 ,4) fucosyltransferase enzyme comprises a UA948 FucTa gene or a strain DMS6709 ...

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan. 114-. (canceled)15. A method for producing insoluble poly alpha-1 ,3-glucan comprising:a) contacting at least water, sucrose, and an isolated glucosyltransferase enzyme comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:4, whereby insoluble poly alpha-1,3-glucan having at least 90% alpha-1,3 glycosidic linkages is produced; andb) isolating the insoluble poly alpha-1,3-glucan produced in step (a).16. The method of claim 15 , wherein said insoluble poly alpha-1 claim 15 ,3-glucan has a number average degree of polymerization of at least 100.17. The method of claim 15 , wherein said insoluble poly alpha-1 claim 15 ,3-glucan has at least 95% alpha-1 claim 15 ,3 glycosidic linkages.18. The method of claim 17 , wherein said insoluble poly alpha-1 claim 17 ,3-glucan has at least 97% alpha-1 claim 17 ,3 glycosidic linkages.19. The method of claim 18 , wherein said insoluble poly alpha-1 claim 18 ,3-glucan has at least 99% alpha-1 claim 18 ,3 glycosidic linkages.20. The method of claim 19 , wherein said insoluble poly alpha-1 claim 19 ,3-glucan has about 100% alpha-1 claim 19 ,3 glycosidic linkages.21. The method of claim 15 , wherein step (a) further comprises contacting a primer with the water claim 15 , sucrose claim 15 , and glucosyltransferase enzyme.22. The method of claim 21 , wherein the primer comprises dextran.23. The method of claim 15 , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 93% identical to SEQ ID NO:4.24. The method of claim 15 , wherein said ...

GLUCURONOSYLTRANSFERASE, GENE ENCODING SAME AND USE THEREOF

Provided are an enzyme involved in a glycyrrhizin biosynthetic system, a gene of the enzyme and use thereof in order to stably and continuously provide a large amount of glycyrrhizin. Glucuronosyltransferase with an activity of further transferring glucuronic acid to the hydroxy group at the 2-position of glucuronic acid in an oleanane-type triterpenoid monoglucuronide is identified to provide the transferase, a gene for the transferase and use thereof. 1. A polypeptide having an activity to transfer glucuronic acid to the hydroxy group at the 2-position of glucuronic acid in an oleanane-type triterpenoid monoglucuronide.2. The polypeptide according to claim 1 , wherein the oleanane-type triterpenoid monoglucuronide is selected from the group consisting of β-amyrin monoglucuronide claim 1 , 11-oxo-β-amyrin monoglucuronide claim 1 , 30-hydroxy-11-oxo-β-amyrin monoglucuronide claim 1 , 30-hydroxy-β-amyrin monoglucuronide claim 1 , 11-oxoglycyrrhetinic acid monoglucuronide and glycyrrhetinic acid monoglucuronide.3GlycyrrhizaMedicago. The polypeptide according to claim 1 , derived from a plant or a plant.4GlycyrrhizaG. uralensis.. The polypeptide according to claim 3 , wherein the plant is5. The polypeptide according to claim 1 , comprising an amino acid sequence represented by SEQ ID NO: 3.6. The polypeptide according to claim 1 , comprising any of amino acid sequences of the following (a) to (c):(a) an amino acid sequence represented by SEQ ID NO: 4,(b) an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 4 by deletion, replacement or addition of one or several amino acids, and or(c) an amino acid sequence having 80% or more identity with the amino acid sequence represented by SEQ ID NO: 4.7. A polynucleotide encoding the polypeptide according to .8. The polynucleotide according to claim 7 , comprising any of nucleotide sequences of the following (d) to (g):(d) a nucleotide sequence represented by SEQ ID NO: 5,(e) a nucleotide sequence ...

Combinatorial dna library for producing modified n-glycans in lower eukaryotes

The present invention relates to eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The invention provides nucleic acid molecules and combinatorial libraries which can be used to successfully target and express mammalian enzymatic activities such as those involved in glycosylation to intracellular compartments in a eukaryotic host cell. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified oligosaccharides are created or selected. N-glycans made in the engineered host cells have a Man 5 GlcNAc 2 core structure which may then be modified further by heterologous expression of one or more enzymes, e.g., glycosyltransferases, sugar transporters and mannosidases, to yield human-like glycoproteins. For the production of therapeutic proteins, this method may be adapted to engineer cell lines in which any desired glycosylation structure may be obtained.

RECOMBINANT BACILLUS SUBTILIS STRAIN FOR PRODUCING UDP-GLYCOSYLTRANSFERASE AND RECOMBINATION METHOD THEREFOR

The present invention discloses a recombinant strain for producing UDP-glycosyltransferase and a recombination method therefor. The recombination method includes the following steps: chemically synthesizing a UDP-glycosyltransferase gene UGT and linking the UGT with a vector pUC57 to obtain pUC57-UGT, and cloning various promoters; linking the obtained pUC57-UGT and each of the promoters to an expression vector to obtain recombinant plasmids; transforming the obtained recombinant plasmids to host strains, respectively, to obtain recombinant plasmids of the host strains; respectively transforming the obtained recombinant plasmids of the host strains to to obtain recombinant strains; and screening out recombinant strain highly expressing UDP-glycosyltransferase from the obtained recombinant strains.

Enzymatic synthesis of soluble glucan fiber

An enzymatically produced soluble α-glucan fiber composition is provided suitable for use as a digestion resistant fiber in food and feed applications. The soluble α-glucan fiber composition can be blended with one or more additional food ingredients to produce fiber-containing compositions. Methods for the production and use of compositions comprising the soluble α-glucan fiber are also provided.

A RECOMBINANT CHIMERIC PROTEIN FOR SELECTINS TARGETING

The invention discloses a recombinant protein (P-selectin glycoprotein ligand-1 and Neural Retina-specific Leucine Zipper) PSGL-1-NRL chimeric protein comprising a Selectin Binding domain and a non-covalent dimerization domain, which is a leucine zipper and is more preferably the leucine zipper domain of the human or mouse Neural Retina-specific Leucine Zipper. The chimeric protein further comprises a covalent dimerization domain with at least one cysteine suitable to form a disulfide bridge with another chimeric protein to form a homodimer. In the chimeric protein, the PSGL-1 domain corresponds to the extracellular region of Human PSGL-1 and is more preferably the selectin binding region of the mature protein. The chimeric protein is correctly post-translationally modified and is efficiently expressed in a mammalian system. It is sulfated, O-linked glycosylated and sialylated and binds P, E and L selectin, allowing in vivo and in vitro targeting for diagnostic or therapeutic purposes. 1. A recombinant chimeric P-Selectin Glycoprotein Ligand-1 (PSGL-1) protein comprising at least: a selectin Binding domain comprising at least aa 5-16 of SEQ ID NO:11 (mature PSGL-1 sequence) , leucine zipper domain comprising an amino acid sequence at least 90% homologous or identical to aa 187-208 of SEQ ID NO:12 (Neural Retina-specific Leucine Zipper) and a disulfide bonds promoting region.2. The recombinant chimeric PSGL-1 protein according to wherein the selectin Binding domain comprises at least an 1-47 of SEQ ID NO:11 (PSGL-1 sequence).3. The recombinant chimeric PSGL-1 protein according to wherein said leucine Zipper comprises an amino acid sequence at least 90% homologous or identical to aa 181-215 of SEQ ID NO:12.4. The recombinant chimeric PSGL-1 protein according to wherein said disulfide bonds promoting region comprises an amino acid sequence defined by the following general formula:{'br': None, 'sub': 1', '2', '3, 'i': n', 'm, '(X)-C(X)-(X)'} [{'sub': 1', '2, 'X, ...

FAGOPYRITOL SYNTHASE GENES AND USES THEREOF

The present invention relates to an isolated DNA molecule encoding a fagopyritol synthase. A method for producing a fagopyritol, an insulin mediator, an insulin mediator analogue, an insulin mediator homologue, or an insulin mediator inhibitor is also described. The method includes providing a fagopyritol synthase, providing a substrate comprising a galactosyl donor and a galactosyl acceptor, and combining the fagopyritol synthase with the substrate under conditions effective produce a fagopyritol, an insulin mediator, an insulin mediator analogue, an insulin mediator homologue, or an insulin mediator inhibitor. 1. An isolated nucleic acid molecule encoding a fagopyritol synthase , wherein the nucleic acid moleculeis at least 80% identical to SEQ ID NO:5 by basic BLAST using default parameters analysis.2Fagopyrum esculentum.. The isolated nucleic acid molecule according to claim 1 , wherein the fagopyritol synthase is from3. The isolated nucleic acid molecule according to claim 2 , wherein the nucleic acid molecule has a nucleotide sequence of SEQ ID NO:5.4. The isolated nucleic acid molecule according to claim 2 , wherein the nucleic acid molecule encodes a protein or polypeptide comprising an amino acid sequence of SEQ ID NO:6.5. The isolated nucleic acid molecule according to claim 1 , wherein the encoded fagopyritol synthase retains biological activity similar to a fagopyritol synthase encoded by SEQ ID NO:5.6Fagopyrum esculentum. An isolated nucleic acid molecule encoding a fagopyritol synthase from claim 1 , wherein the nucleic acid molecule(i) is at least 55% identical to SEQ ID NO:5 by basic BLAST using default parameters analysis and or(ii) hybridizes to a complement of the nucleotide sequence of SEQ ID NO:5 under stringent conditions characterized by a hybridization buffer comprising 5×SSC at a temperature of 55° C.7. An expression vector comprising transcriptional and translational regulatory nucleotide sequences operably linked to the nucleic acid ...

A Salmonella Paratyphi A with an O-Antigen Having an Extended Carbohydrate Chain and Use Thereof

The present invention discloses a A with an O-antigen having an extended carbohydrate chain and uses thereof. The method comprises the following steps: inactivating an cld gene encoding an enzyme controlling chain length of O-antigen of a A strain to obtain a A with deletion of cld gene; allowing overexpression of cldgene encoding an enzyme controlling chain length of O-antigen of in A deficient in the cld gene encoding an enzyme controlling chain length of O-antigen, so as to extend carbohydrate chain length of O-antigen. Both of the A O-polysaccharide-recombinant fusion protein conjugate vaccines rCTB4573-OPSand rEPA4573-OPSas prepared by using A with an O-antigen having an extended carbohydrate chain can induce mice to generate specific antibodies against A, and their antibody titers are significantly improved. 1Salmonella typhimuriumSalmonella paratyphi. A recombinant strain , which is obtained by introducing a cldgene encoding an enzyme controlling chain length of O-antigen of into A which is deficient in cld gene encoding an enzyme controlling chain length of O-antigen.2Salmonella typhimurium. The recombinant strain according to claim 1 , characterized in that the enzyme controlling chain length of O-antigen of has an amino acid sequence with at least 90% identity to the amino acid sequence as shown in SEQ ID NO: 2.3Salmonella paratyphi. The recombinant strain according to claim 1 , characterized in that the A which is deficient in cld gene encoding an enzyme controlling chain length of O-antigen is obtained by a method comprising the following steps:(1) preparing a linear targeting DNA fragment 1, which has a nucleotide sequence as shown in SEQ ID NO: 11, and contains a cat gene;{'i': Salmonella paratyphi', 'S. paratyphi, '(2) transforming pKD46 plasmid into a strain, to obtain a recombinant strain designated as /pKD46;'}{'i': S. paratyphi', 'S. paratyphi', 'S. paratyphi', 'S. paratyphi, '(3) inducing expression of Red recombination system in the /pKD46 ...

BRANCHED ALPHA-GLUCAN, ALPHA-GLUCOSYLTRANSFERASE WHICH FORMS THE GLUCAN, THEIR PREPARATION AND USES

The present invention has objects to provide a glucan useful as water-soluble dietary fiber, its preparation and uses. The present invention solves the above objects by providing a branched α-glucan, which is constructed by glucose molecules and characterized by methylation analysis as follows: 1. A process for producing a saccharide composition comprising branched α-glucan , comprising the steps of:allowing an α-glucosyltransferase and cyclomaltodextrin glucanotransferase to act on maltose and/or α-1,4 glucan having a glucose polymerization degree of 3 or higher; andcollecting the formed saccharide composition comprising branched α-glucan;said saccharide composition is characterized in that the content of water-soluble dietary fiber of the saccharide composition, obtained by applying a high performance liquid chromatography method (Enzyme-HPLC method), is 40% (w/w) or higher.2. The process of claim 1 , wherein said saccharide composition has the following characteristics upon methylation analysis:(1) Ratio of 2,3,6-trimethyl-1,4,5-triacetyl-glucitol to 2,3,4-trimethyl-1,5,6-triacetyl-glucitol, which indicates the ratio of glucose residues with α-1,4 linkages to glucose residues with α-1,6 linkages, is in the range of 1:0.6 to 1:4; and(2) Total content of 2,3,6-trimethyl-1,4,5-triacetyl-glucitol and 2,3,4-trimethyl-1,5,6-triacetyl-glucitol, which indicates that the total content of glucose residues with α-1,4 linkages and glucose residues with α-1,6 linkages, is 60% or higher in the partially methylated glucitol acetates.3. The process of claim 2 , wherein said saccharide composition has the following characteristics upon methylation analysis:(3) Content of 2,4,6-trimethyl-1,3,5-triacetyl-glucitol, which indicates that the content of glucose residues with α-1,3 linkage, is 0.5% or higher but less than 10% in the partially methylated glucitol acetates; and(4) Content of 2,4-dimethyl-1,3,5,6-tetraacetyl-glucitol, which indicates that the content of glucose residues ...

UDP-GLYCOSYLTRANSFERASES FROM SOLANUM LYCOPERSICUM

The present invention relates to polypeptides having UDP-Glycosyltransferase activity derived from Solanum lycopersicum and having the amino acid sequence set out in any of SEQ ID NO: 1 to 4 or an amino acid sequence having at least about 30% sequence identity thereto. The application also relates to recombinant hosts comprising a recombinant nucleic acid sequence encoding said polypeptides and uses thereof prepare glycosylated diterpenes, like steviol glycoside. The host cells might comprise further enzymes of the steviol glycoside biosynthesis pathway. 1. A recombinant host comprising a recombinant nucleic acid sequence encoding a polypeptide comprising:a. the amino acid sequence set forth in SEQ ID NO: 1 or an amino acid sequence having at least about 30% sequence identity thereto;b. the amino acid sequence set forth in SEQ ID NO: 2 or an amino acid sequence having at least about 30% sequence identity thereto;c. the amino acid sequence set forth in SEQ ID NO: 3 or an amino acid sequence having at least about 30% sequence identity thereto; ord. the amino acid sequence set forth in SEQ ID NO: 4 or an amino acid sequence having at least about 30% sequence identity thereto.2. A recombinant host according to which is capable of producing a glycosylated diterpene.3. A recombinant host according to which comprises one or more recombinant nucleotide sequence(s) encoding:a polypeptide having ent-copalyl pyrophosphate synthase activity;a polypeptide having ent-Kaurene synthase activity;a polypeptide having ent-Kaurene oxidase activity; and/ora polypeptide having kaurenoic acid 13-hydroxylase activity.4. A recombinant host according to claim 1 , which comprises a recombinant nucleic acid sequence encoding a polypeptide having NADPH-cytochrome p450 reductase activity.5. A recombinant host according to which comprises a recombinant nucleic acid sequence encoding one or more of:(i) a polypeptide having UGT74G1 activity (UGT3 activity);(ii) a polypeptide having UGT85C2 activity ...

PROCESS FOR PRODUCING APLHA-1,3-GLUCAN POLYMER WITH REDUCED MOLECULAR WEIGHT

A process for producing poly alpha-1,3-glucan with reduced molecular weight is disclosed. The process comprises contacting water, sucrose, a polar organic solvent, and a glucosyltransferase enzyme in a solution to produce poly alpha-1,3-glucan. This contacting step results in the production of poly alpha-1,3-glucan having a reduced molecular weight compared to the molecular weight of a poly alpha-1,3-glucan made in the absence of the polar organic solvent.

METHOD FOR MANUFACTURE OF DEXTRAN, DEXTRAN SOLUTION OBTAINED, AND USES

Microbiological method of production of a dextran solution, according to which a culture medium containing sucrose is inoculated with a preculture of a bacterial strain that is able to produce dextran, then the dextran solution obtained at the end of fermentation is recovered directly, without a subsequent concentration step, characterized in that:—before inoculation, the culture medium contains at least 10 wt. % of sucrose,—after inoculation, sucrose is added again in conditions such that the total amount of sucrose in the medium, including that present before inoculation, is at least 16 wt. %,—the dextran solution obtained contains at least 10 wt. % of dextran. The native solution of dextran obtained and use of the solution as a flocculant. 1. Microbiological method of production of a dextran solution , according to which a culture medium containing sucrose is inoculated with a preculture of a bacterial strain able to produce dextran , then the dextran solution obtained at the end of fermentation is recovered directly , without a subsequent concentration step , characterized in that:before inoculation, the culture medium contains at least 10 wt. % of sucrose,after inoculation, sucrose is added again in conditions such that the total amount of sucrose in the medium, including that present before inoculation, is at least 16 wt. %.2. Method according to claim 1 , characterized in that after inoculation claim 1 , sucrose is added to the medium continuously or discontinuously.3. Method according to claim 1 , characterized in that sucrose is added after inoculation at least twice.4. Method according to claim 1 , characterized in that the total amount of sucrose added claim 1 , including that present in the supplemented medium claim 1 , is at least 20 wt. % claim 1 , preferably at least 25 wt. %.5. Method according to claim 1 , characterized in that the pH is fixed at the start of fermentation at a value of 8 claim 1 , then the reaction is stopped at a value above 5.6. ...

METHOD FOR MANUFACTURING STARCH-CONTAINING FOOD

A starch-containing food with improved properties may be obtained by reacting an actinomycete-derived amylomaltase with starch in the raw material. 1. A method for producing a starch-containing food , comprising reacting an actinomycete-derived amylomaltase with starch in a raw material.2CorynebacteriumStreptomyces.. The production method according to claim 1 , wherein said actinomycete is from the genus or the genus3Corynebacterium glutamicum, Streptomyces avermitilis, Streptomyces cinnamoneus, Streptomyces griseus, Streptomyces thermoviolaceusStreptomyces violaceoruber.. The production method according to claim 1 , wherein said actinomycete is selected from the group consisting of claim 1 , and4. The production method according to claim 1 , wherein said starch-containing food is one or more members selected from the group consisting of a rice processing food claim 1 , a wheat processing food claim 1 , a potato processing food claim 1 , a corn processing food claim 1 , and a tapioca processing food.5. The production method according to claim 1 , wherein said starch-containing food is a processing food containing one or more kinds of starch extracted from rice claim 1 , wheat claim 1 , potato claim 1 , corn claim 1 , or tapioca.6. The production method according to claim 1 , wherein the starch-containing food comprises sucrose.7. A method for producing a starch-containing food product claim 1 , comprising reacting an actinomycete-derived amylomaltase with starch in a raw material claim 1 , wherein said amylomaltase has the amino acid sequence shown in SEQ ID NO: 1 claim 1 , 3 claim 1 , 4 claim 1 , 5 claim 1 , or 6 claim 1 , or an amino acid sequence not less than 90% identical to the amino acid sequence.8. A method for modifying property of a starch-containing food claim 1 , comprising reacting an actinomycete-derived amylomaltase with starch in a raw material.9CorynebacteriumStreptomyces.. The method according to claim 8 , wherein the actinomycete is the genus or ...

RHAMNOLIPID SYNTHESIS

There is provided a method of producing at least one rhamnolipid comprising: 111-. (canceled)13. The method of claim 12 , wherein said organic radicals are alkyl radicals that are optionally branched claim 12 , optionally substituted claim 12 , and optionally unsaturated.14. The method of claim 13 , wherein at least one alkyl radical is hydroxy-substituted.15. The method of claim 13 , wherein at least one alkyl radical is mono- claim 13 , di- or tri-unsaturated.16. The method of claim 12 , wherein said identical or different organic radicals comprise 5 to 13 carbon atoms.17. The method of claim 12 , wherein said carbon source is an alkane selected from the group consisting of: hexane; heptane; octane; nonane; and decane; and/or an alkanoic acid selected from the group consisting of: hexanoic acid; haptanoic acid; octanoic acid; nonanoic acid; and decanoic acid.18. The method of claim 12 , wherein the recombinant cell has been genetically modified such that claim 12 , compared to the wild-type cell claim 12 , the recombinant cell has an increased activity of enzyme E claim 12 , wherein Eis an oxidoreductase.19. The method according to claim 18 , wherein the oxidoreductase is selected from the group consisting of: alkB-type oxidoreductase; monooxygenase; and NAD(P)H dependent alcohol dehydrogenase (ADH).20. The method of claim 19 , wherein the carbon source is hexane and/or decane.21. The method of claim 12 , wherein at least 40% by weight of the total carbon content in the medium is hexane claim 12 , decane claim 12 , hexanoic acid and/or decanoic acid.22. The method of claim 12 , wherein:{'sub': '1', 'a) enzyme Eis able to catalyse the conversion of 3-hydroxyalkanoyl-ACP via 3-hydroxyalkanoyl-3-hydroxyalkanoic acid-ACP to hydroxyalkanoyl-3-hydroxyalkanoic acid;'}{'sub': '2', 'b) enzyme Eis able to catalyse the conversion of dTDP-rhamnose and 3-hydroxyalkanoyl-3-hydroxyalkanoate to α-L-rhamnopyranosyl-3-hydroxyalkanoyl-3-hydroxyalkanoate; and'}{'sub': '3', 'c) enzyme ...

COMPREHENSIVE SINGLE MOLECULE ENHANCED DETECTION OF MODIFIED CYTOSINES

The subject invention provides a method of determining whether a cytosine at a predefined position within a single strand of a double-stranded DNA of known sequence is hydroxymethylated. 1. A method of determining whether a cytosine at a predefined position within a single strand of a double-stranded DNA of known sequence is hydroxymethylated comprising:a) contacting the double-stranded DNA with a glucosyltransferase and a uridine diphosphate glucose (UDP-glucose) so as to replace the hydrogen of hydroxymethylated cytosine with the glucose if the cytosine is hydroxymethylated; andb) determining whether the cytosine contains the glucose;wherein if the cytosine contains the glucose the cytosine is hydroxymethylated cytosine.2. A method of determining whether a cytosine at a predefined position within a single strand of a double-stranded DNA of known sequence is unmethylated comprising:a) treating the double-stranded DNA with an oxidizing agent so as to convert methylated cytosine into hydroxymethylated cytosine if cytosine is methylated;b) contacting the treated double-stranded DNA from step a) with a glucosyltransferase and a uridine diphosphate glucose (UDP-glucose) so as to replace the hydrogen of the hydroxymethylated cytosine with the glucose if the cytosine is hydroxylated; andc) determining whether the cytosine contains the glucose;wherein if the cytosine does not contain glucose the cytosine is unmethylated.3. The method of claim 2 , wherein oxidizing agent is ten-eleven translocation methylcytosine dioxygenase 1 (TET1).4. The method of any of any one of - claim 2 , wherein the glucosyltransferase is T4 β-glucosyltransferase.5. The method of any one of - claim 2 , wherein the glucose is labeled with a detectable chemical group.7. A method of determining whether a cytosine at a predefined position within a single strand of a double-stranded DNA of known sequence is methylated but not hydroxymethylated comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, ...

GELLING DEXTRAN ETHERS

Compositions are disclosed herein comprising at least one dextran ether compound that comprises uncharged, anionic, and/or cationic organic groups. The degree of substitution of one or more dextran ether compounds is about 0.0025 to about 3.0. Dextran from which the disclosed ether compounds can be derived can have a weight-average molecular weight of about 50-200 million Daltons and/or a z-average radius of gyration of about 200-280 nm. Also disclosed are methods of producing dextran ether compounds, as well as methods of using these ether compounds in various applications. 115-. (canceled)16. A composition comprising a dextran compound , wherein the dextran compound is a product of an isolated glucosyltransferase enzyme comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:1 or SEQ ID NO:2 , and wherein the weight-average molecular weight (Mw) of said dextran compound is 50 million to 200 million Daltons.17. The composition of claim 16 , wherein the z-average radius of gyration of the dextran compound is 200-280 nm.18. The composition of claim 16 , wherein the dextran compound comprises chains linked together within a branching structure claim 16 , wherein said chains are similar in length and comprise substantially alpha-1 claim 16 ,6-glucosidic linkages.19. The composition of claim 18 , wherein the average length of the chains is 10-50 monomeric units.20. The composition of claim 16 , wherein the dextran compound comprises:(i) 87-91.5 wt % glucose linked at positions 1 and 6;(ii) 0.1-1.2 wt % glucose linked at positions 1 and 3;(iii) 0.1-0.7 wt % glucose linked at positions 1 and 4;(iv) 7.7-8.6 wt % glucose linked at positions 1, 3 and 6; and (a) positions 1, 2 and 6, or', '(b) positions 1, 4 and 6., '(v) 0.4-1.7 wt % glucose linked at21. The composition of claim 20 , wherein the dextran compound comprises:(i) 89.5-90.5 wt % glucose linked at positions 1 and 6;(ii) 0.4-0.9 wt % glucose linked at positions 1 and 3;(iii) 0.3-0.5 wt % ...

Simplified Process For Producing Maltodextrin and Specialty Syrups

Disclosed are compositions and methods relating to a simplified process for producing maltodextrin and specialty syrups using fewer enzymes and less complicated conditions than required for contemporary enzymatic processes. 1. A method for producing a maltodextrin and/or a specialty syrup comprising contacting a starch substrate with an α-amylase (EC 3.2.1.1) capable of producing , in the substantial absence of a maltogenic enzyme selected from the group consisting of maltogenic amylase (EC 3.2.1.133) , β-amylase (EC 3.2.1.2) , pullulanase (EC 3.2.1.41) , glucoamylase (EC 3.2.1.3) and combinations , thereof , a syrup comprising a DE profile equivalent to the DE profile produced by conventional , multi-enzyme , acid pretreatment conditions that includes a maltogenic enzyme , wherein the method substantially obviates at least one pH adjustment or temperature adjustment step in an otherwise identical process utilizing a different , conventional liquifying α-amylase.2. The method of claim 1 , performed in the absence of a maltogenic enzyme claim 1 , with the exception of the α-amylase claim 1 , which may have maltogentic amylase activity.3. The method of claim 2 , performed in the absence of any maltogenic enzyme claim 2 , with the exception of the α-amylase claim 2 , which may have maltogentic amylase activity.4. The method of any of - claim 2 , wherein the process step is selected from the group consisting of reducing the pH of a liquefact to inactivate a different claim 2 , conventional liquifying α-amylase claim 2 , cooling the liquefact to promote optimal performance of a maltogenic enzyme claim 2 , heating a saccharified liquifact to inactivate the maltogenic enzyme claim 2 , and cooling the saccharified liquifact to concentrate the product.5Cytophaga. The method of any of - claim 2 , wherein the α-amylase is from a sp.6Cytophaga. The method of any of - claim 2 , wherein the α-amylase is the α-amylase from sp. having the amino acid sequence of SEQ ID NO: 1 claim 2 ...

A MULTIFUNCTIONAL RECOMBINANT NUCLEOTIDE DEPENDENT GLYCOSYLTRANSFERASE PROTEIN AND ITS METHOD OF GLYCOSYLATION THEREOF

The present invention generally relates to a method of peptides' or polypeptides' modification by glycosylation. In particular, the invention relates to one pot synthesis of disaccharide glycan on to the acceptor substrate and thereby generating O- and/or S-glycosylated neo-glycopeptides including antimicrobial peptides by using multifunctional recombinant nucleotide dependent glycosyltransferase. 1. An in vitro one pot method for synthesis of O-linked and/or S-linked diglycosylated products , the method comprising:a. providing a mixture of a donor substrate and an acceptor substrate in a ratio in the range of 20:1 to 400:1, wherein the donor substrate is an activated nucleotide sugar selected from the group consisting of saccharide-UDP, saccharide-GDP, and related nucleotide sugars thereof and the acceptor substrate comprises a peptide or polypeptide having amino acid sequence selected from the group consisting of SEQ ID NO.: 4, SEQ ID NO.: 5, SEQ ID NO.: 6, SEQ ID NO.: 7, SEQ ID NO.: 8, SEQ ID NO.: 9, SEQ ID NO.: 10, SEQ ID NO.: 11, SEQ ID NO.: 12, SEQ ID NO.: 13, SEQ ID NO.: 14, SEQ ID NO.: 15, SEQ ID NO.: 16 and SEQ ID NO.: 17;b. providing a multifunctional recombinant nucleotide dependent diglycosyltransferase protein having amino acid sequence selected from the group consisting of SEQ ID NO.: 1, SEQ ID NO.: 2, and SEQ ID NO.: 3;c. contacting a multifunctional recombinant nucleotide dependent diglycosyltransferase protein obtained in step (b) with the mixture obtained in step (a);d. reacting the donor substrate and the acceptor substrate in presence of the multifunctional recombinant nucleotide dependent diglycosyltransferase protein, wherein the glycosyltransferase protein catalyzes the transfer of plurality of saccharide moieties from the donor substrate to serine, threonine or cysteine residue in the acceptor substrate to obtain the O-linked and/or S-linked di-glycosylated products.2. The method of claim 1 , wherein the donor substrate saccharide-UDP sugar ...

MODIFIED POLYNUCLEOTIDES FOR TREATING PROTEIN DEFICIENCY

The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules. 1. A method of producing a polypeptide of interest in a mammalian cell or tissue comprising contacting said mammalian cell or tissue with a modified mRNA , wherein said modified mRNA encodes a member selected from the group consisting of serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1254) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1255) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1256) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1257) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1258) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1259) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1260) , SERPINA1 (SEQ ID NO: 1261) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1262) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1263) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1264) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1265) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1266) , serpin peptidase inhibitor , clade A (alpha-1 antiproteinase , antitrypsin) , member 1 (SEQ ID NO: 1267) , arginase , liver (SEQ ID NO: 822) , arginase , liver (SEQ ID NO: 823) , arginase , liver (SEQ ID NO: 824) , argininosuccinate lyase (SEQ ID NO: 825) , argininosuccinate lyase ( ...

ANTIGEN BINDING MOLECULES WITH INCREASED Fc RECEPTOR BINDING AFFINITY AND EFFECTOR FUNCTION

The present invention relates to antigen binding molecules (ABMs). In particular embodiments, the present invention relates to recombinant monoclonal antibodies, including chimeric, primatized or humanized antibodies specific for human CD20. In addition, the present invention relates to nucleic acid molecules encoding such ABMs, and vectors and host cells comprising such nucleic acid molecules. The invention further relates to methods for producing the ABMs of the invention, and to methods of using these ABMs in treatment of disease. In addition, the present invention relates to ABMs with modified glycosylation having improved therapeutic properties, including antibodies with increased Fc receptor binding and increased effector function. 173-. (canceled)74: A host cell engineered to express at least one nucleic acid encoding a polypeptide having β(1 ,4)-N-acetylglucosaminyltransferase III activity in an amount sufficient to modify the oligosaccharides in the Fc region of a polypeptide produced by said host cell , wherein said polypeptide is an antigen binding molecule comprising a sequence derived from the muring B-Ly1 antibody and a sequence from a heterologous polypeptide.75: The host cell of claim 74 , wherein said polypeptide having β(1 claim 74 ,4)-N-acetylglucosaminyltransferase III activity is a fusion polypeptide.76: The host cell of claim 74 , wherein said antigen binding molecule is an antibody.77: The host cell of claim 74 , wherein said antigen binding molecule is an antibody fragment.78: The host cell of claim 74 , wherein said antigen binding molecule comprises a region equivalent to the Fc region of a human IgG.79: The host cell of claim 74 , wherein said antigen binding molecule produced by said host cell exhibits increased Fc receptor binding affinity as a result of said modification.80: The host cell of claim 74 , wherein said antibody produced by said host cell exhibits increased effector function as a result of said modification.81: The host cell ...

QUANTITATIVE CONTROL OF SIALYLATION

The present disclosure is directed to the use of certain glycosyltransferase variants having N-terminal truncation deletions. Contrary to previous findings certain truncations were found to exhibit sialidase enzymatic activity, particularly a variant of human sialyltransferase (hST6Gal-I) with a truncation deletion involving the first 89 N-terminal amino acids of the respective wild-type polypeptide. A fundamental finding documented in the present disclosure is that there exists a variant of this enzyme which is capable of catalyzing transfer of a glycosyl moiety as well as hydrolysis thereof. Thus, disclosed is a specific exemplary variant of mammalian glycosyltransferase, nucleic acids encoding the same, methods and means for recombinantly producing the variant of mammalian glycosyltransferase and use thereof, particularly for sialylating in a quantitatively controlled manner terminal acceptor groups of glycan moieties being part of glycoproteins such as immunoglobulins. 1. A method to hydrolyze the α2 ,6 glycosidic bond in a N-acetylneuraminyl-α2 ,6-β-D-galactosyl-1 ,4-N-acetyl-β-D-glucosamine moiety , the moiety being a terminal structure of a glycan in a sialylated glycoprotein or glycolipid , the method comprising the steps of(a) providing in an aqueous solution a sialylated glycoprotein or glycolipid with a terminal N-acetylneuraminyl-α2,6-β-D-galactosyl-1,4-N-acetyl-β-D-glucosamine moiety in the glycan portion of said glycoprotein;(b) incubating the N-acetylneuraminyl-α2,6-β-D-galactosyl-1,4-N-acetyl-β-D-glucosamine moiety with a N-terminally truncated human β-galactoside-α-2,6-sialyltransferase I having the amino acid sequence of SEQ ID NO:2;thereby hydrolyzing the α2,6 glycosidic bond in the N-acetylneuraminyl-α2,6-β-D-galactosyl-1,4-N-acetyl-β-D-glucosamine moiety.2. The method according to claim 1 , wherein step (a) is performed under conditions permitting glycosyltransferase enzymatic activity.3. The method according to claim 1 , wherein step (b) is ...

A METHOD FOR INCREASING RESISTANT STARCH AND DIETARY FIBER IN RICE

The present invention discloses mutations in the genes encoding starch synthases and also in starch branching enzymes associated with enhanced dietary fibre and resistant starch levels in the endosperm of a suitable variety of rice. The dietary fiber and resistant starch are enhanced to an extent to significantly reduce the hydrolysis index values of the rice grains to 35%-40%. These rice varieties are in great demand for diabetic population and provide a number of other health benefits such as reduced body weight gain, cardiac health and colon health. As this strategy does not involve the use of genetic manipulation technologies, it can be directly employed in the rice breeding programmers without any restrictions. 1. A rice plant comprising one or more mutations in a combination of two , three or four genes that includes SSI , SS IIIa , SBE I and SBE IIb; wherein said rice plant produces seed that germinates , and further wherein grain from said rice plant has an increased resistant starch or total dietary fibre level as compared to grain from a wild type rice plant.2. The rice plant of claim 1 , further comprising a reduced levels of enzymes Starch Synthase I and/or Starch Synthase IIIa and in combination with reduced levels of Starch Branching Enzyme I and/or Starch Branching Enzyme IIb in starch granules resulting from mutations in a combination of two claim 1 , three or four genes coding these enzymes of said plant as compared to starch granules of a wild type rice plant.3. The rice plant of claim 1 , wherein starch of the grain has an enhanced amylose content of more than 26% as compared to the grains of the wild type rice plant.4. The rice plant of claim 1 , wherein starch in the grains has an enhanced resistant starch content of more than 6% as compared to the grains of wild type rice plant.5Oryza sativa. The rice plant of which is of race indica type.6. Rice grain from the rice plant of .7. Flour comprising a cell of the rice grain of .8. A food or ...

COMPOSITIONS AND METHODS COMPRISING THE USE OF EXIGUOBACTERIUM ACETYLICUM AND BACILLUS COAGLUANS ALPHA-GLUCANOTRANSFERASE ENZYMES

An isolated and/or purified α-glucanotransferase from , recombinantly engineered variants thereof, active fragments thereof, synthetic nucleic acids encoding the α-glucanotransferase and variants thereof, host cells comprising the synthetic nucleic acids, and compositions comprising the α-glucanotransferase are provided. Methods of using the compositions include the manufacture of oligosaccharides. 126-. (canceled)27. A composition comprising at least:(i) an isolated GH70 subfamily 4 α-glucanotransferase comprising an amino acid sequence that is at least 95% identical to amino acids 33-736 of SEQ ID NO:7 or amino acids 22-726 of SEQ ID NO:11, and(ii) a glucooligosaccharide with α-(1→4) and α-(1→6) glycosidic linkages.28. The composition of claim 27 , wherein the α-glucanotransferase comprises an amino acid sequence that is at least 98% identical to amino acids 33-736 of SEQ ID NO:7 or amino acids 22-726 of SEQ ID NO:11.29. The composition of claim 27 , wherein the α-glucanotransferase comprises an amino acid sequence that is at least 99% identical to amino acids 33-736 of SEQ ID NO:7 or amino acids 22-726 of SEQ ID NO:11.30. The composition of claim 27 , wherein the α-glucanotransferase comprises an amino acid sequence that is at least 99.5% identical to amino acids 33-736 of SEQ ID NO:7 or amino acids 22-726 of SEQ ID NO:11.31. The composition of claim 27 , wherein the α-glucanotransferase comprises amino acids 31-731 of SEQ ID NO:2 or amino acids 22-726 of SEQ ID NO:11.32. The composition of claim 27 , wherein the composition is in a lyophilized powder form claim 27 , an encapsulated form claim 27 , a coated form claim 27 , a granulated form claim 27 , or a liquid formulation.33. The composition of claim 27 , further comprising a diluent.34. The composition of claim 27 , wherein the α-glucanotransferase comprises said amino acid sequence that is at least 95% identical to amino acids 33-736 of SEQ ID NO:7.35. The composition of claim 27 , wherein the α- ...

PROCESS FOR THE MANIPULATION OF NUCLEIC ACIDS

The present invention discloses a process for engineering a host cell comprising the steps of; a) integrating a first polynucleotide cassette including a first selection marker flanked by a first pair of recombination sites; b) removing the first selection marker by the action of a recombinase which recognises the first pair of recombination sites; c) integrating a second polynucleotide cassette including a second selection marker flanked by a second pair of recombination sites; and d) removing the second selection marker by the action of a recombinase which recognises the second pair of recombination sites. Also disclosed is a host cell genome polynucleotide comprising a first recombinantly engineered region and a second recombinantly engineered region, wherein a first single recombination site is adjacent to the first recombinantly engineered region, and a second single recombination site is adjacent to the second recombinantly engineered region. 125.-. (canceled)26. A method of removing at least two portions of insert nucleic acid from a genomic polynucleotide in a host cell , said method comprising the steps of:a) preparing the genomic polynucleotide comprising a first insert nucleic acid which is flanked by a pair of first recombination sites in the same orientation which are identical to each other and have a first nucleic acid sequence;b) exposing the genomic polynucleotide of step a) to a recombinase that recognises the first recombination sites such that the identical recombination sites recombine resulting in the excision of the first insert nucleic acid and one of the first recombination sites;c) inserting into the genomic polynucleotide of step b) a second insert nucleic acid flanked by a pair of second recombination sites in the same orientation wherein the second recombination sites are identical to each other and have a second nucleic acid sequence which shares 70-98% sequence identity with the first nucleic acid sequence; andd) exposing the genomic ...

Compositions and Methods for Functionalizing and Linking Materials

The invention relates to methods for functionalizing a material, linking materials, or producing a composite material with compositions comprising (a) a xyloglucan endotransglycosylase, a polymeric xyloglucan, and a functionalized xyloglucan oligomer; (b) a xyloglucan endotransglycosylase, a polymeric xyloglucan functionalized with a chemical group, and a functionalized xyloglucan oligomer; (c) a xyloglucan endotransglycosylase, a polymeric xyloglucan functionalized with a chemical group, and a xyloglucan oligomer; (d) a xyloglucan endotransglycosylase, a polymeric xyloglucan, and a xyloglucan oligomer; (e) a xyloglucan endotransglycosylase and a polymeric xyloglucan functionalized with a chemical group; (f) a xyloglucan endotransglycosylase and a polymeric xyloglucan; (g) a xyloglucan endotransglycosylase and a functionalized xyloglucan oligomer; (h) a xyloglucan endotransglycosylase and a xyloglucan oligomer, or (i) a composition of (a), (b), (c), (d), (e), (f), (g), or (h) without a xyloglucan endotransglycosylase. The invention also relates to a material obtained by such methods. 1. A method for functionalizing a material comprising treating the material with a composition selected from the group consisting of (a) a composition comprising a xyloglucan endotransglycosylase , a polymeric xyloglucan , and a functionalized xyloglucan oligomer comprising a chemical group; (b) a composition comprising a xyloglucan endotransglycosylase , a polymeric xyloglucan functionalized with a chemical group , and a functionalized xyloglucan oligomer comprising a chemical group; (c) a composition comprising a xyloglucan endotransglycosylase , a polymeric xyloglucan functionalized with a chemical group , and a xyloglucan oligomer; (d) a composition comprising a xyloglucan endotransglycosylase , a polymeric xyloglucan , and a xyloglucan oligomer; (e) a composition comprising a xyloglucan endotransglycosylase and a polymeric xyloglucan functionalized with a chemical group; (f) a ...

PRODUCTION OF MODIFIED GLYCOPROTEINS HAVING MULTIPLE ANTENNARY STRUCTURES

The present invention relates to lower eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar and sugar nucleotide transporters to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified lipid-linked oligosaccharides are created or selected. N-glycans made in the engineered host cells exhibit GnTIV, GnTV, GnT VI or GnTIX activity, which produce multiantennary N-glycan structures and may be modified further by heterologous expression of one or more enzymes, e.g., glycosyltransferases, sugar, sugar nucleotide transporters, to yield human-like glycoproteins. For the production of therapeutic proteins, this method may be adapted to engineer cell lines in which any desired glycosylation structure may be obtained. 1. A process for making a glycoprotein in a lower eukaryotic host cell comprising the step of introducing into the cell an N-acetylglucosaminyltransferase activity selected from the group consisting of: N-acetylglucosaminyltransferase IV activity , an N-acetylglucosaminyltransferase V activity , an N-acetylglucosaminyltransferase VI activity; and an N-acetylglucosaminyltransferase IX activity wherein the glycoprotein comprises at least three antennae on the trimannose core.2. A process for making a glycoprotein in a lower eukaryotic host cell comprising the step of expressing in the cell one or more enzymatic activities that produce multiple antennary N-glycans comprising GlcNAcβ1 ,2-Manα1 ,6 (GlcNAcβ1 ,4 GlcNAcβ1 ,2 Manα1 ,3) Man β1 ,4-GlcNAcβ1 ,4-GlcNAcβ1 ,4-Asn; GlcNAcβ1 ,4 GlcNAcβ1 ,2-Manα1 ,6 (GlcNAcβ1 ,4 GlcNAcβ1 ,2 Manα1 ,3) Man β1 ,4-GlcNAcβ1 ,4-GlcNAcβ1 ,4-Asn; and GlcNAcβ1 ,6 GlcNAcβ1 ,4 GlcNAcβ1 ,2-Manα1 ,6 (GlcNAcβ1 ,4 GlcNAcβ1 ,2 Manα1 ,3) Man β1 ,4-GlcNAcβ1 ,4- ...

Engineered glucosyltransferases

Disclosed herein are glucosyltransferases with modified amino acid sequences. Such engineered enzymes synthesize alpha-glucan products having increased molecular weight. Further disclosed are reactions and methods in which engineered glucosyltransferases are used to produce alpha-glucan.

Engineered glucosyltransferases

Disclosed herein are glucosyltransferases with modified amino acid sequences. Such engineered enzymes exhibit improved alpha-glucan product yields and/or lower leucrose yields, for example. Further disclosed are reactions and methods in which engineered glucosyltransferases are used to produce alpha-glucan.

NOVEL GLYCOSYLTRANSFERASE DERIVED FROM DOLWOE AND USE THEREOF

Provided are a novel UDP-glycosyltransferase (uridine diphosphate glycosyltransferase) protein having glycosyltransfer activity for glucose linked by a glycosidic bond at the C-20 position of PPD (protopanaxadiol)-type or PPT (protopanaxatriol)-type ginsenoside, and use thereof. 1. A method of preparing a PPD or PPT-type ginsenoside which is glycosylated at glucose linked by a glycosidic bond at the C-20 position using a UDP-glycosyltransferase protein having a glycosyltransfer activity for glucose linked by a glycosidic bond at the C-20 position of PPD (protopanaxadiol)-type or PPT (protopanaxatriol)-type ginsenoside; a vector comprising a polynucleotide encoding the protein; a transformant introduced with the vector; or a culture of the transformant.2. The method of claim 1 , wherein the PPD or PPT-type ginsenoside having glucose linked by a glycosidic bond at the C-20 position is one or more selected from the group consisting of CK (compound K) claim 1 , F2 claim 1 , Rd claim 1 , F1 claim 1 , Rgand Re.3. The method of claim 1 , wherein preparation of the glycosylated ginsenoside comprises one or more selected from the group consisting of conversion of CK into gypenoside LXXV claim 1 , conversion of F2 into gypenoside XVII claim 1 , conversion of Rd into Rb claim 1 , conversion of F1 into notoginsenoside U claim 1 , conversion of Rginto notoginsenoside R3 claim 1 , and conversion of Re into gluco-Re.4. The method of claim 1 , wherein the UDP-glycosyltransferase protein is defined by an amino acid sequence of SEQ ID NO: 1.5. A method of glycosylation of a PPD (protopanaxadiol)-type or PPT (protopanaxatriol)-type ginsenoside having glucose linked by a glycosidic bond at the C-20 position claim 1 , comprising administering a UDP-glycosyltransferase protein having a glycosyltransfer activity for glucose linked by a glycosidic bond at the C-20 position of PPD-type or PPT-type ginsenoside;a vector comprising a polynucleotide encoding the protein; a transformant ...

PREBIOTIC COMPOSITION OR PHARMACEUTICAL COMPOSITION SYNTHESIZED FROM CATALYTIC DOMAINS PRODUCING HIGHLY ALPHA-1,2 BRANCHED DEXTRAN

The invention relates to an isolated polypeptide with an glycosyl transferase enzymatic activity for producing dextrans with .alpha.(1.fwdarw.2) sidechains, comprising at least one region for bonding to glucan and a catalytically active region situated beyond the region bonding to glucan. The invention further relates to polynucleotides coding for said enzymes and vectors containing the same. 1. A prebiotic composition comprising 'a suitable carrier.', 'an isolated polypeptide having an enzymatic glycosyltransferase activity capable of forming dextrans having α(1→2) linkages from saccharose, α-D-fluoroglucose, paranitrophenyl-α-D glucopyranoside, α-D-glucopyranoside-α-D sorbofuranoside or 4-O-α-D galactopyranosylsucrose, wherein said isolated polypeptide has at least one glucan binding domain and a catalytic activity domain of SEQ ID NO: 1 located downstream of the glucan binding domain; and'}2. The prebiotic composition according to claim 1 , wherein the polypeptide has at least two catalytic domains located either side of the glucan binding domain.3. The prebiotic composition according to claim 1 , wherein the polypeptide has a peptide signal claim 1 , a variable zone claim 1 , two catalytic domains and a glucan binding domain located between the two catalytic domains.4. The prebiotic composition according to claim 2 , wherein at least one of the two catalytic domains has/have a percentage similarity in the range of 65% to 100% with SEQ ID NO:1.5. The prebiotic composition according claim 1 , wherein the size of the glucan binding domain is more than 500 amino acids.6. The prebiotic composition according to claim 5 , wherein the polypeptide contains SEQ ID NO:2.7. The prebiotic composition according to claim 6 , modified by substitution claim 6 , insertion or deletion of amino acid sequences and comprising sequences having at least 90% similarity with the following sequences of SEQ ID No.2: 423-439 (SEQ ID No:6) 2120-2138 (SEQ ID No:12) 478-501 (SEQ ID No:7) 2161- ...

Alpha (1,2) Fucosyltransferase Syngenes For Use in the Production of Fucosylated Oligosaccharides

The invention provides compositions and methods for engineering or other host production bacterial strains to produce fucosylated oligosaccharides, and the use thereof in the prevention or treatment of infection. 1. A method for producing a fucosylated oligosaccharide in a bacterium comprising providing bacterium comprising an exogenous lactose-utilizing α(1 ,2) fucosyltransferase enzyme , wherein the amino acid sequence of said enzyme comprises at least 22% but less than 40% identity to FutC (SEQ ID NO: 1).2. A method for producing a fucosylated oligosaccharide in a bacterium comprising providing bacterium comprising an exogenous lactose-utilizing α(1 ,2) fucosyltransferase enzyme , wherein the amino acid sequence of said enzyme comprises at least 25% identity to FutN (SEQ ID NO: 3).3LachnospiraceaeTannerellaClostridium bolteaeMethanosphaerula palustriesAkkermansia muciniphiliaClostridium bolteae. The method of or , wherein said α(1 ,2) fucosyltransferase enzyme comprises , sp. FutQ , sp. FutS , +13 FutP , FutR , FutY , FutP , or a functional variant or fragment thereof.4. The method of claim 1 , wherein said α(1 claim 1 ,2) fucosyltransferase enzyme comprises any one of amino acid sequences SEQ ID NO: 10-21 and 292 claim 1 , or a functional variant or fragment thereof.5. The method of claim 1 , further comprising retrieving the fucosylated oligosaccharide from said bacterium or from a culture supernatant of said bacterium.6. The method of claim 1 , wherein said fucosylated oligosaccharide comprises 2′-fucosyllactose (2′-FL) claim 1 , lactodifucotetraose (LDFT) claim 1 , Lacto-N-fucopentaose I (LNF I) claim 1 , or lacto-N-difucohexaose I (LDFH I).7. The method of claim 1 , wherein the bacterium further comprises an exogenous lactose-utilizing α(1 claim 1 ,3) fucosyltransferase enzyme and/or an exogenous lactose-utilizing α(1 claim 1 ,4) fucosyltransferase enzyme.8Helicobacter pylori. The method of claim 7 , wherein the exogenous lactose-utilizing α(1 claim 7 ,3) ...

Alpha (1,2) Fucosyltransferases Suitable for Use in the Production of Fucosylated Oligosaccharides

The invention provides compositions and methods for engineering or other host production bacterial strains to produce fucosylated oligosaccharides, and the use thereof in the prevention or treatment of infection. 1. A nucleic acid construct comprising an isolated nucleic acid encoding a lactose-utilizing α(1 ,2) fucosyltransferase enzyme , said nucleic acid being operably linked to one or more heterologous control sequences that direct the production of the enzyme in a host bacteria production strain , wherein the amino acid sequence of said enzyme encoded by said nucleic acid comprises at least 10% and less than 40% identity to SEQ ID NO:2.2Eschericia coli. The construct of claim 1 , wherein said production strain comprises K12.3. The construct of claim 1 , wherein said nucleic acid encodes a WbgL claim 1 , FutL claim 1 , or FutN protein claim 1 ,4. The construct of claim 1 , wherein said heterologous control sequence comprises a bacterial promoter and operator claim 1 , a bacterial ribosome binding site claim 1 , a bacterial transcriptional terminator claim 1 , or a plasmid selectable marker.5Bacillus. The construct of claim 1 , wherein said production strain is a member of the genus.6Bacillus licheniformis.. The construct of claim 1 , wherein said production strain comprises7Pantoea. The construct of claim 1 , wherein said production strain is a member of the genus.8LactobacillusLactococcus. The construct of claim 1 , wherein said production strain is a member of the or genus.9Streptococcus, Proprionibacterium, Enterococcus, Bifidobacterium, Sporolactobacillus, Micromomospora, Micrococcus, RhodococcusPseudomonas. The construct of claim 1 , wherein said production strain is a member of the claim 1 , or genus.10Bacillus licheniformis, Bacillus subtilis, Bacillus coagulans, Bacillus thermophilus, Bacillus laterosporus, Bacillus megaterium, Bacillus mycoides, Bacillus pumilus, Bacillus lentus, Bacillus cereusBacillus circulans, Erwinia herbicolaPantoea ...

ENZYMATICALLY PRODUCED CELLULOSE

Enzymatic reactions are disclosed herein comprising water, glucose-1-phosphate, cellodextrin, and at least one cellodextrin phosphorylase enzyme comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:2 or SEQ ID NO:6. These reactions produce a low molecular weight, insoluble cellulose with enhanced features. 112-. (canceled)13. An isolated composition comprising:(i) insoluble cellulose having a weight-average degree of polymerization (DPw) of at least 15,(ii) a cellodextrin phosphorylase enzyme comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:2 or SEQ ID NO:6, and(iii) a starch phosphorylase or sucrose phosphorylase.14. The isolated composition of claim 13 , wherein the insoluble cellulose has a DPw of at least 17.15. The isolated composition of claim 13 , wherein the insoluble cellulose has a DPw of 15 to 50.16. The isolated composition of claim 13 , wherein the insoluble cellulose has a DPw of 15 to 30.17. The isolated composition of claim 13 , wherein the insoluble cellulose has a DPw of 17 to 50.18. The isolated composition of claim 13 , wherein the insoluble cellulose has a DPw of 17 to 30.19. The isolated composition of claim 13 , wherein the cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:6.20. The isolated composition of claim 14 , wherein the cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:6.21. The isolated composition of claim 15 , wherein the cellodextrin phosphorylase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:2 or SEQ ID NO:6.22. The isolated composition of claim 13 , wherein the isolated composition comprises the sucrose phosphorylase.23. The isolated composition of claim 13 , wherein the isolated composition comprises the starch phosphorylase.24. The isolated composition of claim 23 , wherein the isolated ...