ПРОИЗВОДСТВО МОЛОЧНЫХ ОЛИГОСАХАРИДОВ ЧЕЛОВЕКА В МИКРОБНЫХ ПРОДУЦЕНТАХ С ИСКУССТВЕННЫМ ИМПОРТОМ/ЭКСПОРТОМ

Изобретение относится к производству лакто-N-триозы II генетически модифицированной клеткой микроорганизма-хозяина. Способ производства лакто-N-триозы II генетически модифицированной клеткой микроорганизма-хозяина включает получение генетически модифицированную клетку микроорганизма-хозяина, которая включает по меньшей мере одну рекомбинантную β-1,3-N-ацетилглюкозаминилтрансферазу, повышенную экспрессию или активность по меньшей мере одного белка, осуществляющего экспорт сахара, способного осуществлять экспорт лакто-N-триозы II. Указанную клетку культивируют и получают лакто-N-триозу II из среды. При этом указанный по меньшей мере один белок, осуществляющий экспорт сахара, выбран из группы, состоящей из YjhB из Escherichia coli, ProP из Mannheimia succiniciproducens, SetA из Cedecea neteri и их функциональных фрагментов. Предложена также генетически модифицированная клетка микроорганизма-хозяина для производства лакто-N-триозы II. Изобретение обеспечивает улучшенный экспорт и получение ...

СПОСОБ ПОЛУЧЕНИЯ РЕБАУДИОЗИДА M С ИСПОЛЬЗОВАНИЕМ ФЕРМЕНТАТИВНОГО СПОСОБА

Группа изобретений относится к биотехнологии. Предложен способ получения ребаудиозида М. Способ включает взаимодействие ребаудиозида D с донором глюкозила в присутствии UDP-глюкозилтрансферазы или рекомбинантных клеток, содержащих UDP-глюкозилтрансферазу, с получением ребаудиозида M. При этом донором глюкозила является UDP-глюкоза или система регенерации UDP-глюкозы, включающая сахарозу, сахарозасинтетазу и UDP, UDP-глюкозилтрансфераза имеет аминокислотную последовательность, имеющую по меньшей мере 90% идентичность SEQ ID NO:2. Рекомбинантные клетки представляют собой микробные клетки, выбранные из группы, состоящей из клеток Escherichia coli, Saccharomyces cerevisiae или Pichia pastoris. Предложен также способ получения ребаудиозида М, включающий взаимодействие ребаудиозида А с донором глюкозила в присутствии первой UDP-глюкозилтрансферазы или рекомбинантных клеток, содержащих первую UDP-глюкозилтрансферазу с образованием ребаудиозида D, и взаимодействие ребаудиозида D с донором глюкозила ...

СИНТЕЗ НМО

Изобретение относится к биотехнологии, в частности к бактериальной клетке, стабильно культивируемой в среде, где клетка адаптирована для продукции олигосахаридов. Причем клетка трансформирована таким образом, что она содержит по меньшей мере одну последовательность нуклеиновых кислот, кодирующую фермент, задействованный в синтезе олигосахарида. При этом дополнительно клетка трансформирована таким образом, что она содержит по меньшей мере одну последовательность нуклеиновых кислот, кодирующую белок семейства эффлюксного переносчика сахара или его функциональный гомолог. Кроме того, изобретение относится к способу продукции олигосахаридов, задействующего вышеуказанную клетку. Изобретение позволяет получать олигосахариды с высокой степенью эффективности. 2 н. и 11 з.п. ф-лы, 3 ил., 2 табл., 4 пр.

СПОСОБ ОБРАБОТКИ КРАХМАЛА

... 1. Одностадийный способ получения растворимого гидролизата крахмала, где способ включает одновременное воздействие на водную суспензию зернистого крахмала при температуре ниже начальной температуры желатинизации указанного зернистого крахмала: первого фермента, который: (a) является членом семейства 13 гликозидгидролазы; (b) имеет альфа-1,4-глюкозидную гидролитическую активность; и (c) включает углевод-связанный модуль семейства 20, и по меньшей мере одного второго фермента, который является бета-амилазой (EC 3.2.1.2), или глюкоамилазой (EC 3.2. 1.3). 2. Способ по п.1, где суспензия крахмала имеет 20-55% сухого вещества зернистого крахмала, предпочтительно 25-40% сухого вещества зернистого крахмала, более предпочтительно 30-35% сухого вещества, особенно предпочтительно приблизительно 33% сухого вещества зернистого крахмала. 3. Способ по п.1, где по меньшей мере 85%, 86%, 87%, 88%,89% 90%, 91%, 92%, 93% в наименьшей степени 94%, 95%, 96%, 97%, 98% или по меньшей мере 99% сухого вещества ...

СИНТЕЗ НОВЫХ ПРОИЗВОДНЫХ СИАЛООЛИГОСАХАРИДОВ

... 1. Способ синтеза соединений общей формулы 1A и их солейгде одна из групп R представляет собой α-сиалильную группировку, а другая представляет собой H, Xозначает углеводный линкер, A представляет собой D-глюкопиранозильное звено, необязательно замещенное фукозилом, Rпредставляет собой защитную группу, которая может быть удалена гидрогенолизом, целое число m равно 0 или 1, отличающийся тем, что сиалил-донор формулы SA-ORили его соль, где Rможет представлять собой моно-, ди- или олигосахарид, гликолипид, гликопротеин или гликопептид, циклическую или ациклическую алифатическую группу или арильный остаток, a SA представляет собой α-сиалильную группировку, вводят в реакцию с сиалил-акцептором общей формулы 2A или его солью,где X, A, m и Rявляются такими, как определено выше, в условиях катализа ферментом, обладающим транс-сиалидазной активностью.2. Способ по п.1, в котором фермент, обладающий транс-сиалидазной активностью, выбран из сиалидаз, получаемых из видов Bifidobacterium, и транс-сиалидаз ...

СИАЛИЛТРАНСФЕРАЗЫ И ИХ ПРИМЕНЕНИЕ В ПОЛУЧЕНИИ СИАЛИРОВАННЫХ ОЛИГОСАХАРИДОВ

СПОСОБ ПОЛУЧЕНИЯ КОМПОЗИЦИИ ЧАСТИЦ, СОДЕРЖАЩЕЙ БЕЗВОДНУЮ КРИСТАЛЛИЧЕСКУЮ 2-О-α-D-ГЛЮКОЗИЛ-L-АСКОРБИНОВУЮ КИСЛОТУ

... 1. Способ получения композиции в форме частиц, содержащей безводную кристаллическую 2-O-α-D-глюкозил-L-аскорбиновую кислоту, включающий следующие стадии (a)-(e):(a) обеспечение воздействия цикломальтодекстринглюканотрансферазы на раствор, содержащий либо разжиженный крахмал, либо декстрин и L-аскорбиновую кислоту в качестве исходных материалов, и затем обеспечение воздействия глюкоамилазы на полученный раствор с получением раствора, содержащего 2-O-α-D-глюкозил-L-аскорбиновую кислоту, с выходом продуцирования 2-O-α-D-глюкозил-L-аскорбиновой кислоты по меньшей мере 27%;(b) очистка полученного раствора, содержащего 2-O-α-D-глюкозил-L-аскорбиновую кислоту, с получением содержания 2-O-α-D-глюкозил-L-аскорбиновой кислоты более 86%, в расчете на массу сухого твердого вещества;(c) осаждение безводной кристаллической 2-O-α-D-глюкозил-L-аскорбиновой кислоты из очищенного раствора с содержанием 2-O-α-D-глюкозил-L-аскорбиновой кислоты более 86%, в расчете на массу сухого твердого вещества, способом ...

СЛИТНЫЕ ФЕРМЕНТЫ

... 1. Рекомбинантный белок, обладающий активностью N-ацетилглюкозаминилтрансферазы, при этом рекомбинантный белок катализирует перенос N-ацетилглюкозамина на концевой остаток Manα3 и катализирует перенос N-ацетилглюкозамина на концевой остаток Manα6 акцепторного гликана, и при этом рекомбинантный белок содержит каталитический домен, по меньшей мере, из двух разных ферментов.2. Рекомбинантный белок по п.1, в котором рекомбинантный белок представляет собой слитый белок, содержащий каталитический домен N-ацетилглюкозаминилтрансферазы I и каталитический домен N-ацетилглюкозаминилтрансферазы II.3. Рекомбинантный белок по п.2, в котором каталитический домен N-ацетилглюкозаминилтрансферазы I и каталитический домен N-ацетилглюкозаминилтрансферазы II происходят из ферментов человека.4. Рекомбинантный белок по п.3, в котором каталитический домен N-ацетилглюкозаминилтрансферазы I содержит последовательность, которая по меньшей мере, на 70%, по меньшей мере, на 75%, по меньшей мере, на 80%, по меньшей ...

СПОСОБ ИНГИБИРОВАНИЯ ИЗОМЕРИЗАЦИИ ВОССТАНАВЛИВАЮЩЕГО САХАРИДА ПРИ ТЕПЛОВОЙ ОБРАБОТКЕ

СИНТЕЗ НМО

... 1. Клетка, стабильно культивируемая в среде, где клетка адаптирована для продукции олигосахаридов, клетка трансформирована таким образом, что она содержит по меньшей мере одну последовательность нуклеиновых кислот, кодирующую фермент, задействованный в синтезе олигосахарида,которая характеризуется тем, чтоклетка дополнительно трансформирована таким образом, что она содержит по меньшей мере одну последовательность нуклеиновых кислот, кодирующую белок семейства эффлюксного переносчика сахара, ее функциональный гомолог или производное.2. Клетка в соответствии с п.1, отличающаяся тем, что клетку выбирают из группы, включающей клетки бактерий, грибов, животных и растений.3. Клетка в соответствии с п.2, отличающаяся тем, что клетка представляет собой клетку Escherichia coli.4. Клетка в соответствии с любым из пп.1-3, отличающаяся тем, что фермент выбирают из группы, включающей гликозилтрансферазу, гликозилтрансферазу Leloir типа, гликозилтрансферазу не-Leloir типа, фукозилтрансферазу, сиалилтрансферазу ...

Verfahren zur enzymatischen Herstellung von Alpha-Glykosiden und deren Estern

KALORIENARMES SUESSUNGSMITTEL UND VERFAHREN ZUR HERSTELLUNG VON NAHRUNGSMITTELN UND GETRAENKEN MIT NIEDRIGEM KALORIENGEHALT UNTER VERWENDUNG DESSELBEN

Oligosaccharide prodn. in high yield and selectivity

Prodn. of oligosaccharides comprises reacting glucosyl donors with glucosyl acceptors in the presence of glucosyl-transferases at a temp. below the freezing point of the reaction mixt.

POLYPEPTIDE

Amylose granule and its preparation

Novel amylose granules advantageously used in the fields of food products, pharmaceuticals and cosmetics exist in an approximately globular-shape of amylose granule or in a conjugation form consisting of two or more of the amylose granules linked together, and having about 2.10 mu m in diameter or major axis, B type form of starch on powder X-ray diffraction analysis, the number-average molecular weight of about 4,000-7,000 on gel permeation chromatography, and the weight- average molecular weight per the number-average molecular weight of about 1.4-1.7. The granules may be prepared by allowing cyclomaltodextrin glucanotransferase (EC 2.4.1.19) to act on an aqueous solution containing a cyclodextrin or starch. In examples the enzyme is derived from Bacillus stearothermophilas or Bacillus macerans; other sources are mentioned. ...

Enzymatic production of sucrose-6-ester, an intermediate for the manufacture of halo sugars

A novel process is described for production of 6-acyl-sucrose comprising enzymatic acylation of sucrose by an esterifying agent including an organic acid in presence of a lipase or an esterase in a solvent in which the enzyme used is stable. Chlorinated sucrose, the high intensity sweetener trichlorogalactosucrose can be prepared by chlorination and deacylation of 6-acyl sucrose prepared by the process of this invention.

CRYSTALLINE ERLOSE HYDRATE

PROCESSES FOR PRODUCING SYRUPS OR SYRUP SOLIDS CONTAINING FRUCTOSE-TERMINATED OLIGOSACCHARIDES

Regioselective glycosylation

Mapping cytosine modifications

Methods, compositions and kits for selectively altering and detecting modified cytosine residues are provided.

PRODUCTION OF ISOMALTULOSE

Recombinant microorganisms expressing an oligosaccharide receptor mimic

Chemeric carbohydrates produced by recombinant microorganism carrying exogenous glycosyl trans ferases act with or without exogenous enzymes required for synthesis or nucleotide synthesis precursors. These recombinant microorganism can be used as a means for competitively inhibiting the binding of toxins or adhesins to receptors of mucosal surfaces, especially gas-trointestinal surface. In particular chimeric sugar moieties have been made for lipopolysaccharides, in recombinant microorganism that present multiple copies of the oligosaccharides. The oligosaccharide moieties so presented act as receptor mimic for toxins and adhesins. A number have been synthesised and have been shown to confer protection against attach by pathogenic organisms or their products in vitro and an in vivo.

Recombinant microorganisms expressing an oligosaccharide receptor mimic.

Saccharide compositions methods and apparatus for their synthesis

Recombinant microorganisms expressing an oligosaccaride receptor mimic

Recombinant microorganisms expressing an oligosaccaride receptor mimic

FUCOSYLTRANSFERASE-GEN

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.

TRANSGLYKOSYLIERUNGSVERFAHREN AND THE PRODUCTION OF A GLYKOPROTEINS

POLYPEPTID WITH ALPHAISOMALTOSYLGLUCOSACCHARIDSYNTHASE ACTIVITY

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

PROCEDURE FOR THE PRODUCTION OF OLIGOSACCHARIDEN

PROCEDURE FOR THE ENZYMATIC PRODUCTION OF FRUCTOSYLVERBINDUNGEN

PROCEDURE FOR THE PRODUCTION OF CYCLODEXTRIN

PROCEDURE FOR THE ENZYMATIC PRODUCTION OF ALPHA GLYKOSIDEN AND THEIR ESTERS

PRODUCTION OF ISOMALTULOSE.

NICHTREDUZIERENDE OLIGOSACCHARIDE AND YOUR PRODUCTION AND USE

VARIANTS OF THE CYCLOMALTODEXTRIN GLUCANOTRANSFERASE

ALPHA-2,8/2,9-POLYSIALYLTRANSFERASE

BY AROMATICS SUBSTITUTED GLYKOSID

METHOD FOR THE ISOLATION OF GLYKOSYLTRANSFERASEN

ALPHA AMYLASE RESISTANT POLYSACCHARIDE, PROCEDURES FOR THEIR PRODUCTION, USE, AND FOOD CONTAINING THESE POLYSACCHARIDE

OLIGOSACCHARIDE AS ENZYME SUBSTRATES AND - INHIBITORS: PROCEDURE AND COMPOSITIONS

Engineered beta-glucosidases and glucosylation methods

The present invention provides engineered β-glucosidase (BGL) enzymes, polypeptides having BGL activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. The present invention also provides compositions comprising the enzymes, and methods of using the engineered BGL enzymes to make products with β-glucose linkages.

In vivo synthesis of sialylated compounds

The present invention is in the technical field of synthetic biology and metabolic engineering. More particularly, the present invention is in the technical field of fermentation of metabolically engineered microorganisms. The present invention describes engineered microorganisms able to synthesize sialylated compounds via an intracellular biosynthesis route. These microorganisms can dephosphorylate N-acetylglucosamine-6-phosphate to N- acetylglucosamine and convert the N-acetylglucosamine to N- acetylmannosamine. These microorganisms also have the ability to convert N-acetylmannosamine to N-acetyl-neuraminate. Furthermore, the present invention provides a method for the large scale ...

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.

Production of complex carbohydrates

HIGH PURITY GENTIOOLIGOSACCHARIDES OBTAINED THEREFROM AND USES THEREOF

The present invention relates to methods for preparing high purity gentiooligosaccharides, high purity gentiooligosaccharides obtained therefrom, and uses thereof. The methods of the present invention involve: adding a low purity gentiooligosaccharide to a liquid medium; subjecting the liquid medium to inoculation with a microorganism, followed by incubation and fermentation to consume glucose that is contained in the low purity gentiooligosaccharide; and subjecting the resulting fermentation broth to filtration and purification. According to the method of the present invention, high purity gentiooligosaccharides having a purity of at least 90% that can be used as alternatives to foods such as cocoa, chocolate, coffee, beer, tea, bread or confectionery product, and beverage or the main ingredients thereof.

Manipulation of plant cellulose

Substitutes for modified starch in paper manufacture

Alpha (1,2) fucosyltransferases suitable for use in the production of fucosylated oligosaccharides

The invention provides compositions and methods for engineering ...

Alpha (1,3) fucosyltransferases for use in the production of fucosylated oligosaccharides

The invention relates to methods and compositions for the production of fucosylated oligosaccharides.

Alpha (1,2) fucosyltransferase syngenes for use in the production of fucosylated oligosaccharides

The invention provides compositions and methods for engineering ...

Production of isomaltooligosaccharides and uses therefor

... - 27 Described herein are methods for preparing isomaltooligosaccharides ("IMOs") from a carbohydrate substrate and uses thereof. In the presence of a maltogenic enzyme and additional IMO precursors, an Aspergillus sp. invertase is 5 capable of producing IMOs from a starch slurry. The ability of the invertase to function as a transglucosidase enzyme imparts a simultaneous mechanism for IMO saccharification. 17/08/11,19417 speci.27 ...

Non-caloric sweeteners and methods for synthesizing

Disclosed are steviol glycosides referred to as rebaudioside V and rebaudioside W. Also disclosed are methods for producing rebaudioside M (Reb M), rebausoside G (Reb G), rebaudioside KA (Reb KA), rebaudioside V (Reb V) and rebaudioside (Reb W). The present disclosure relates generally to natural sweeteners. More particularly, the present disclosure relates to non-caloric sweeteners and methods for synthesizing the non-caloric sweeteners.

Methods and materials for biosynthesis of mogroside compounds

Methods and materials for biosynthesis of mogroside compounds are described.

Polysaccharide fibers

Food for diabetics

Alpha-amylase resistant polysaccharides, production method and use thereof and food products containing said polysaccharides

Method for producing water-insoluble alpha-1,4-glucan

GLUCO-OLIGOSACCHARIDE MIXTURE AND A PROCESS FOR ITS MANUFACTURE

METHOD OF CONVERTING STARCH TO (BETA)-CYCLODEXTRIN

PROCESS FOR PRODUCING .ALPHA.1,4-GALACTOSYLTRANSFERASE AND GALACTOSE-CONTAINING COMPLEX SUGAR

A process for producing a protein having an .alpha.1,4-galactosyltransferase activity with the use of a transformant containing a recombinant DNA having a DNA encoding a protein having an .alpha.1,4-galactosyltransferase activity originating in a microorganism belonging to the genus Pasteurella, and a galactose-containing complex sugar with the use of a transformant producing a protein having an .alpha.1,4-galactosyltransferase activity originating in a microorganism.

CONVERSION OF SUCROSE TO FRUCTOSE AND ETHANOL

BRANCHED CYCLODEXTRINS AND METHODS OF PREPARING THEM

Disclosed is a novel mannosyl-cyclodextrin having a mannosyl group bonded to a hydroxyl group of a glucosyl group of a cyclodextrin via an .alpha.-bond and a method of producing the mannosyl-cyclodextrin. The new branched cyclodextrin is expected to be widely useful in various fields of drug industry, food industry and cosmetic industry.

PROCESSES FOR PREPARING IMMOBILIZED ENZYME AND REACTION PRODUCT WITH SAID IMMOBILIZED ENZYME

The present invention relates to processes for preparing immobilized enzyme, particularly immobilized-starch-degrading enzyme, and reaction product with said immobilized enzyme. More precisely, the present processes are based on the invention that an immobilized-starch-degrading enzyme having a high starch-degrading activity is easily obtainable by modifying in solution a starch-degrading enzyme with a modifying reagent in a manner that the enzyme is not insolubilized substantially, and adsorbing the modified enzyme onto a carrier. The processes are also based on the finding that reaction products from amylaceous materials can be easily produced with said immobilizedstarch-degrading enzyme.

ALPHA 1,2-FUCOSYLTRANSFERASE FROM HELICOBACTER PYLORI

A bacterial .alpha.1,2-fucosyltransferase gene and deduced amino acid sequence is provided. The gene is useful for preparing .alpha.1,2-fucosyltransferase polypeptide, and active fragment thereof, which can be used in the production of oligosaccharides such as Lewis X, Lewis Y, Lewis B and H type 1, which are structurally similar to certain tumor-associated carbohydrate antigens found in mammals. These product glycoconjugates also have research and diagnostic utility in the development of assays to detect mammalian tumors.

UDP-DEPENDENT GLYCOSYLTRANSFERASE FOR HIGH EFFICIENCY PRODUCTION OF REBAUDIOSIDES

Provided herein are compositions and methods for improved production of steviol glycosides in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding a Setaria italica UDP-glycosyltransferase 40087 or its variant UDP-glycosyltransferase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding a UDP-glycosyltransferase sr.UGT_9252778, Bd_UGT10850, and/or Ob_UGT91B1_like. In some embodiments, the host cell further comprises one or more heterologous nucleotide sequence encoding further enzymes of a pathway capable of producing steviol glycosides in the host cell. The compositions and methods described herein provide an efficient route for the heterologous production of steviol glycosides, including but not limited to, rebaudioside D and rebaudioside M.

METHOD FOR PRODUCING TREHALOSE EMPLOYING A TREHALOSE PHOSPHORYLASE VARIANT

The present invention relates to a method for producing trehalose, comprising the steps of mixing and reacting, in any order, (i) at least one alpha-phosphorylase capable of catalyzing the production of alpha-D-glucose 1-phosphate intermediate from a saccharide raw material, and from at least one phosphorus source;(ii) at least one trehalose phosphorylase capable of catalyzing the production of trehalose from an alpha-D-glucose 1-phosphate intermediate and a glucose substrate, wherein the trehalose phosphorylase is a trehalose phosphorylase variant with an amino acid sequence which differs from the amino acid sequence of a wild type trehalose phosphorylase in at least one amino acid position, (iii) at least one saccharide raw material which produces an alpha-D-glucose 1-phosphate intermediate and a co-product by catalytic action of the alpha-phosphorylase; and (iv) at least one phosphorus source selected from the group consisting of a phosphoric acids and an inorganic salt thereof.

METHODS OF MAKING SYRUPS

Provided herein are methods of making a substantially clear syrup comprising reacting one or more substrates at a concentration of at least 40 % w/w with at least one alternan sucrase enzyme at a temperature of greater than 45~C.

LARGE SCALE ENZYMATIC SYNTHESIS OF OLIGOSACCHARIDES

A novel UDP-Gal regeneration process and its combined use with a galactosyltransferease 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.

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.

PROCESS FOR THE MONO- AND BI-SIALYLATION OF GLYCOPROTEINS EMPLOYING N-TERMINALLY TRUNCATED BETA-GALACTOSIDE ALPHA-2,6-SIALYLTRANSFERASE MUTANTS

The present disclosure is directed to the use of certain glycosyltransferase variants having N-terminal truncation deletions. It was found that the combination of two different truncation variants of human ß-galactoside-a-2,6-sialyltransferase I (hST6Gal-I) exhibited different specific sialyltransferase enzymatic activities. In one example, under conditions wherein the first variant ?89 hST6Gal-I catalyzed formation of bi-sialylated target molecules the second variant ?108 hST6Gal-I catalyzed formation of mono-sialylated target molecules. Thus, disclosed are variants of mammalian glycosyltransferase, nucleic acids encoding the same, methods and means for recombinantly producing the variants 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.

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.

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 ...

BOVINE MILK OLIGOSACCHARIDES

Oligosaccharides from bovine milk, whey and dairy products, and methods of producing bovine milk oligosaccharides are provided.

BOVINE MILK OLIGOSACCHARIDES

Oligosaccharides from bovine milk, whey and dairy products, and methods of producing bovine milk oligosaccharides are provided.

FOOD CONTAINING FRUCTOSE

HEAT-RESISTANT .BETA.-GALACTOSYLTRANSFERASE, ITS PRODUCTION PROCESS AND ITS USE

Described herein are a novel heat-resistant p-galactosyltransferase, a production process of the enzyme and a utilization method of the enzyme. The enzyme is produced preferably by a microorganism belonging to the family of Acrinomycetaceae, which may be selected from fungi belonging to the genus Saccharopolyspora, the genus Thermomonospora or the genus Thermoactinomyces.

HEAT RESISTANT MALTOSE PHOSPHORYLASE, PROCESS FOR PREPARATION THEREOF, BACTERIA USED FOR PREPARATION THEREOF, AND METHODS FOR USING THE ENZYME

This invention relates to heat resistant maltose phosphorylase having an activity of 80 % or more of the one untreated after treated in a buffer of pH 6.0, at one temperature of 50 to 60.degree.C for 15 minutes, a process for preparation thereof, bacteria used for preparation thereof, and processes for preparation of .beta.-glucose-1 -phosphoric and trehalose using the enzyme. By carrying out enzymatic reaction at high reaction temperatures using this enzyme, it is possible to prepare .beta.-glucose-1-phosphoric acid or trehalose industrially advantageously, with lowering of contamination with various germs and shortening of reaction time.

MANIPULATION OF CELLULOSE AND/OR .BETA.-1,4-GLUCAN

The present invention relates generally to isolated genes which encode polypeptides involved in cellulose biosynthesis in plants and transgenic plants expressing same in sense or antisense orientation, or as ribozymes, cosuppression or gene-targeting molecules. More particularly, the present invention is directed to a nucleic acid molecule isolated from Arabidopsis thaliana, Oryza sativa, wheat, barley, maize, Brassica ssp., Gossypium hirsutum and Eucalyptus ssp. which encode an enzyme which is important in cellulose biosynthesis, in particular the cellulose synthase enzyme and homologues, analogues and derivatives thereof and uses of same in the production of transgenic plants expressing altered cellulose biosynthetic properties.

SUBSTITUTES FOR MODIFIED STARCH IN PAPER MANUFACTURE

The present invention provides methods of making paper utilizing glucans, produced by the glucosyltransferase C enzyme of the species Streptococcus mutans, instead of modified starches. The present glucans are functionally similar to the hydroxethyl modified starch and are particularly useful in th e coating step of paper manufacture. The present glucans also exhibit thermoplastic properties and impart gloss to the paper during the coating st ep.

IMPROVED NUCLEIC ACIDS FOR REDUCING CARBOHYDRATE EPITOPES

The invention relates to nucleic acids which encode a first glycosyltransferase which competes with a second enzyme for a substrate, thereby reducing the formation of a product of the second enzyme. The nucleic acids are useful in producing cells and organs with reduced antigenicity and which may be used for transplantation.

Cyclodextrins complexing agents for foodstuffs tobacco

Improved method for cyclodextrins (I). Complexing agents e.g. for foodstuffs, tobacco, and medicinals. A starch hydrolysate with dextrose equiv. (DE) is not > 20 & pref. 0.5-6 is incubated with cyclodextrin transglycosylase in an aqueous medium for 2-7 days at 40-60 deg. & pH 5.5-7.5. I is isolated by pptn. with a complexing agent, e.g. PhBr, C2H2-Cl4, etc. or by crystallization. a) High working concn. (20-50% solids) vs. 5% by former methods. b) Byproduct formation minimised.

Procédé de préparation d'un agent édulcorant à base de saccharose et d'amidon

PROCEDE POUR LA PREPARATION D'UN EDULCORANT.

Verfahren zur Herstellung von Laevanen praktisch einheitlicher Molekülgrössen

Procédé pour la production d'un édulcorant

СПОСОБ ПОЛУЧЕНИЯ САХАРОЗЫ-6-АЦЕТАТА ПУТЕМ ЦЕЛЬНОКЛЕТОЧНОГО БИОКАТАЛИЗА

Приведено описание способа, в котором используются иммобилизированные или не иммобилизированные цельноклеточные препараты микроорганизма, включающего Aureobasidium pullulans, способного образовывать ферменты группы фруктозилтрансферазы для катализирования реакции между сахарозой и 6-0O-защищенной глюкозой для образования 6-O-защищенной сахарозы, промежуточного химического соединения при синтезе трихлоргалактосахарозы. 6-O-защищенную сахарозу отделяют от высокомолекулярных побочных продуктов реакции, имеющих молекулярный вес 500 Да и выше, путем обратного осмоса и далее очищают путем колончатой хроматографии.

СОЗДАНИЕ ХИМИЧЕСКИХ СТРОИТЕЛЬНЫХ БЛОКОВ ИЗ РАСТИТЕЛЬНОЙ БИОМАССЫ ПОСРЕДСТВОМ ИЗБИРАТЕЛЬНОЙ ДЕПОЛИМЕРИЗАЦИИ

Изобретение относится к способу ферментативной обработки необработанного полимерного сырья, включающему следующие стадии: (а) предпочтительно отделение растворимых компонентов от необработанного полимерного сырья, (b) обработку необработанного полимерного сырья ферментативной системой для высвобождения определенных растворимых мономерных или олигомерных строительных блоков из нерастворимого необработанного полимерного сырья; и (с) отделение определенных мономерных или олигомерных строительных блоков, полученных на стадии b), из остатков необработанного полимерного сырья. Предпочтительно, ферментативная система, используемая на стадии b), обладает не более чем 50%, предпочтительно не более чем 20%, более предпочтительно не более чем 10%, более предпочтительно не более чем 5%, более предпочтительно не более чем 2%, более предпочтительно не более чем 1% других ферментативных активностей, помимо ферментативной активности, приводящей к высвобождению указанных определенных мономерных или олигомерных ...

STRAIN OF BACTERIUM BACILLUS CIRCULANS B-65 û PRODUCER OF CYCLOMALTODEXTRINGLUKANOTRANSFERASE, METHOD FOR ISOLATION THEREOF AND USE FOR THE PREPARATION OF ?-CYCLODEXTRIN

HETERO-TRANSGLYCOSYLASE AND USES THEREOF

RECOMBINANT OF GLYCOSYL TRANSFERASE AND METHODS GLYCOSYLATION STEVIOLOVYKh GLYCOSIDES

СОЗДАНИЕ ХИМИЧЕСКИХ СТРОИТЕЛЬНЫХ БЛОКОВ ИЗ РАСТИТЕЛЬНОЙ БИОМАССЫ ПОСРЕДСТВОМ ИЗБИРАТЕЛЬНОЙ ДЕПОЛИМЕРИЗАЦИИ

Изобретение относится к способу ферментативной обработки необработанного полимерного сырья, включающему следующие стадии: а) предпочтительно отделение растворимых компонентов от необработанного полимерного сырья, b) обработку необработанного полимерного сырья ферментативной системой для высвобождения определенных растворимых мономерных или олигомерных строительных блоков из нерастворимого необработанного полимерного сырья и с) отделение определенных мономерных или олигомерных строительных блоков, полученных на стадии b), из остатков необработанного полимерного сырья. Предпочтительно ферментативная система, используемая на стадии b), обладает не более чем 50%, предпочтительно не более чем 20%, более предпочтительно не более чем 10%, более предпочтительно не более чем 5%, более предпочтительно не более чем 2%, более предпочтительно не более чем 1% других ферментативных активностей, помимо ферментативной активности, приводящей к высвобождению указанных определенных мономерных или олигомерных ...

METHOD OF PRODUCING 1 - KESTOZY

METHOD OF EXTRACTION OF BETAINE FROM MOLASSES

METHODS AND MEANS OF APPLYING COATING ON PAPER BY APPLICATION OF FILM COATING

ENZYMATIC GLYCOSYLATION STEVIOLGLIKOZIDOV AND OTHER COMPOUNDS WITH GLUCOSE - 1 - PHOSPHATE

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.

Food product containing starch gel, starch granule, production method and use thereof

Here is provided a method of producing a starch gel-containing food, the method comprising the steps of: treating starch granules with an enzyme at a temperature of about 10° C. or higher and about 70° C. or lower to obtain an enzyme-treated starch; mixing a food material, the enzyme-treated starch and water to obtain a mixture; heating the mixture thereby gelatinizing the enzyme-treated starch in the mixture; and cooling the mixture containing the gelatinized enzyme-treated starch thereby gelling the starch to obtain a starch gel-containing food, wherein the enzyme is selected from the group consisting of amyloglucosidase, isoamylase, α-glucosidase, α-amylase having a characteristic capable of improving a gel forming ability of a starch, and cyclodextrin glucanotransferase.

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 ...

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 ...

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 ...

Process for producing a particulate composition comprising anhydrous crystalline 2-o-alpha-d-glucosyl-l-ascorbic acid

The invention provides a process for enabling the production of a particulate composition containing anhydrous crystalline ascorbic acid 2-glucoside that does not significantly cake even when the production yield of ascorbic acid 2-glucoside does not reach 35% by weight. The process for producing a particulate composition containing anhydrous crystalline ascorbic acid 2-glucoside, which comprises allowing a CGTase to act on a solution containing either liquefied starch or dextrin and L-ascorbic acid and then allowing a glucoamylase to act on the resulting solution to obtain a solution with an ascorbic acid 2-glucoside production yield of at least 27%, purifying the obtained solution to increase the ascorbic acid 2-glucoside content to a level of over 86% by weight, precipitating anhydrous crystalline ascorbic acid 2-glucoside by a controlled cooling method or pseudo-controlled cooling method, collecting the precipitated anhydrous crystalline ascorbic acid 2-glucoside, and ageing and drying the collected anhydrous crystalline ascorbic acid 2-glucoside.

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.

Glucosylated steviol glycosides as a flavor modifier

A taste and flavor profile modifying composition is described. The composition includes glucosylated steviol glycosides which can modify the intensity of a taste and/or a flavor in a food or beverage product.

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.

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 ...

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 , ...

Glucosylated steviol glycosides composition as a flavor modifier

A taste and flavor profile enhancing composition includes glucosylated steviol glycosides which can enhance the intensity of a taste and/or a flavor in a food or beverage product. A method of increasing the taste and flavor intensity of a food or beverage product, includes the step of adding a taste and flavor enhancing composition to the food or beverage product, wherein the taste and flavor enhancing composition includes glucosylated steviol glycosides. A method of improving the organoleptic properties of a food or beverage product including a high fructose syrup, including the step of adding the taste and flavor enhancing composition to the food or beverage product. Adding the taste and flavor enhancing composition may cause the high fructose syrup, such as high fructose corn syrup, to taste more like sugar.

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 ...

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 ...

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.

SEPARATION OF OLIGOSACCHARIDES FROM FERMENTATION BROTH

The present invention relates to the isolation and purification of sialylated oligosaccharides from an aqueous medium in which they are produced. 1. A method for separating a sialylated oligosaccharide from an aqueous medium , the method comprising treating an aqueous medium containing said sialylated oligosaccharide with a strong anion exchange resin in Cl-form and a strong cation exchange resin.2. The method of claim 1 , wherein said aqueous medium is a fermentation broth or an enzymatic reaction mixture containing said sialylated oligosaccharide.3. The method of claim 1 , wherein the separated sialylated oligosaccharide is obtained in the form of its alkali metal salt.4. The method of claim 1 , wherein the strong cation exchange resin is in an alkali metal cation form or H-form.5. The method of claim 1 , further comprising neutralizing an eluate of the strong cation resin in H-form with an alkali metal containing basic solution.6. The method of claim 1 , wherein said treating of said aqueous medium with said strong cation exchange resin follows treating said aqueous medium with said strong anion exchange resin.7. The method of claim 2 , wherein said fermentation broth is obtained by culturing a genetically modified cell claim 2 , wherein said cell is capable of producing said sialylated oligosaccharide from an internalized carbohydrate precursor.8. The method of claim 1 , wherein said sialylated oligosaccharide is a sialylated lactose.9. The method of claim 8 , wherein said sialylated lactose is 3′-SL or 6′-SL.1012-. (canceled)13. The method of claim 6 , wherein said treating of said aqueous medium with said strong cation exchange resin immediately follows treating said aqueous medium with said strong anion resin.14. The method of claim 7 , wherein said genetically modified cell is from a genetically modified microorganism.15E. coli,E. coli. The method of claim 14 , wherein said genetically modified microorganism is an wherein the contains one or more ...

Compositions and methods for making alpha-(1,2)-branched alpha-(1,6) oligodextrans

Compositions for improving the health of a subject comprise alpha-(1,2)-branched alpha-(1,6) oligodextrans, preferably with an average molecular weight between about 10 kDa and 70 kDa, between about 10% and 50% alpha-(1,2)-osidic side chains, and having at least partial indigestibility in the subject. Methods for improving the health of a subject comprise administering the composition to a subject in an amount effective to improve gut health, or to prevent or treat a gastrointestinal disorder, a cholesterol-related disorder, diabetes, or obesity. Methods for making oligodextrans having controlled size and controlled degree of branching comprise providing alpha-(1,6) oligodextrans having an average molecular weight between 0.5 and 100 kDa and introducing at least 10% alpha-(1,2)-osidic side chains onto the alpha-(1,6) oligodextrans.

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- ...

Process for the attachment of a galnac moiety comprising a (hetero)aryl group to a glcnac moiety, and product obtained thereby

The present invention relates to a process for attaching an N-acetylgalactosamine-(hetero)arylmoiety to an N-acetylglucosaminemoiety, the process comprising the step of contacting the N-acetylgalactosamine-(hetero)arylmoiety with the N-acetylglucosaminemoiety in the presence of a mutant galactosyltransferase, wherein the N-acetylglucosaminemoiety is according to Formula (1) the N-acetylgalactosamine-(hetero)arylmoiety is according to Formula (2): In a particularly preferred embodiment of the process according to the invention, the N-acetylgalactosamine-(hetero)arylmoiety comprises a 1,3-dipole functional group, and the N-acetylglucosaminemoiety is a terminal GlcNAc moiety of a glycoprotein glycan. The invention further relates to a product obtainable by the process according to the invention, in particular to glycoproteins. Also, the invention relates to several compounds comprising an N-acetylgalactosamine-(hetero)arylmoiety.

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.

Enzyme Exhibiting a-1,6-Glucosyl Transfer Activity

The present invention relates to an enzyme having α-1,6-glucosyl transfer activity, which can use a partially degraded starch product as a substrate and is heat resistant and suitable for industrial applications; an enzyme preparation for use in manufacturing α-1,6-glucan, comprising the enzyme as an active ingredient; and a method for manufacturing α-1,6-glucan using the enzyme or enzyme preparation. The present invention provides an enzyme having α-1,6-glucosyl transfer activity, which is any one of proteins (a), (b), and (c): (a) a protein consisting of an amino acid sequence of SEQ ID NO: 3; (b) a protein consisting of an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 3; and (c) a protein consisting of an amino acid sequence in which one or several amino acid(s) have been substituted, inserted, deleted and/or added in the amino acid sequence of SEQ ID NO: 3.

Enzymatic Preparation Method of Inclusion Complexes of Tributyrin

The disclosure relates to an enzymatic preparation method of inclusion complexes of tributyrin, and belongs to the technical field of oil microencapsulation. The disclosure combines enzymatic synthesis of cyclodextrin and inclusion of tributyrin with cyclodextrin, including enzymatic preparation of cyclodextrin with a CGT enzyme. Tributyrin is added in the preparation process; after reaction, Tween is added, and homogenization and spray drying are carried out. The effect of the finally obtained tributyrin powder is much better than that of single inclusion of tributyrin with cyclodextrin. The disclosure is simple in process, low in cost and convenient in operation; reaction processes are free of toxicity and pollution; there are no toxic reagent residues; the inclusion effect is obvious, and better utilization of a nutritional additive tributyrin in actual production is facilitated. 1. A method of enzymatic preparation of inclusion complexes of tributyrin , comprising: preparing starch milk or a maltodextrin solution as a raw material , wherein a mass concentration of the starch milk or maltodextrin solution is 15% to 30%; adding an enzyme into the starch milk or maltodextrin solution for liquefaction reaction and cyclization reaction; and after the cyclization reaction is completed , adding tributyrin and an emulsifier so that tributyrin is included.2. The method according to claim 1 , wherein starch in the starch milk is one or more of corn starch claim 1 , waxy corn starch claim 1 , potato starch claim 1 , rice starch claim 1 , cassava starch and wheat starch claim 1 , and maltodextrin in the maltodextrin solution is maltodextrin from a starch source with a DE value of 5 to 25.3. The method according to claim 1 , further comprising: after the adding tributyrin and the emulsifier claim 1 , mixing emulsification claim 1 , shearing claim 1 , homogenization and spray drying.4. The method according to claim 1 , wherein the enzyme is cyclodextrin glucosyltransferase.5. ...

METHOD FOR PRODUCING GALACTOOLIGOSACCHARIDE

Provided is a method for improving the production amount of a tri- or higher galactooligosaccharide and the reaction rate by a method for producing a galactooligosaccharide characterized by allowing β-galactosidase to react with a substrate in the presence of 5 to 60 mM sodium ions and 0.5 to 8 mM magnesium ions. 1. A method for producing a galactooligosaccharide , characterized by allowing β-galactosidase to react with a substrate in the presence of 5 to 60 mM sodium ions and 0.5 to 8 mM magnesium ions.2. The production method according to claim 1 , wherein the concentration of magnesium ions is from 1.5 to 8 mM.3Kluyveromyces.. The production method according to claim 1 , wherein β-galactosidase is derived from a microorganism belonging to the genus The present invention relates to a method for producing a galactooligosaccharide using β-galactosidase, more particularly relates to a method for improving the production amount of a tri- or higher galactooligosaccharide and the reaction rate by allowing β-galactosidase to act on a substrate in the coexistence of specific metal ions at a predetermined concentration.β-Galactosidase is known to catalyze a transgalactosylation reaction as well as a hydrolysis reaction of a β-D-galactoside bond in lactose or the like, and is used in the production of a galactooligosaccharide that selectively allows Bifidobacteria to grow in the intestine.A method for improving a transgalactosylation ratio in such a reaction using β-galactosidase has been studied. For example, a method for increasing the transgalactosylation ratio by increasing the concentration of lactose serving as a substrate and allowing β-galactosidase to act thereon has been proposed (PTL 1).A product obtained by a transgalactosylation reaction using β-galactosidase can include, other than tri- or higher galactooligosaccharides such as β-D-galactopyranosyl-(1-4) β-D-galactopyranosyl-D glucose (4′-GL), for example, transgalactosylated disaccharides such as β-D- ...

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 ...

CHEMOENZYMATIC SYNTHESIS OF HEPARIN AND HEPARAN SULFATE ANALOGS

The present invention provides a one-pot multi-enzyme method for preparing UDP-sugars from simple sugar starting materials. The invention also provides a one-pot multi-enzyme method for preparing oligosaccharides from simple sugar starting materials. 1. A method of synthesizing a UDP-sugar , the method comprising forming a reaction mixture comprising a first sugar , a nucleotide-sugar pyrophosphorylase , and a first enzyme selected from the group consisting of a kinase and a dehydrogenase , under conditions sufficient to form the UDP-sugar.2. The method of claim 1 , wherein the first sugar is selected from the group consisting of substituted or unsubstituted glucose (Glc) claim 1 , substituted or unsubstituted glucose-1-phosphate (Glc-1-P) claim 1 , substituted or unsubstituted glucuronic acid (GlcA) claim 1 , substituted or unsubstituted glucuronic acid-1-phosphate (GlcA-1-P) claim 1 , substituted or unsubstituted iduronic acid (IdoA) claim 1 , substituted or unsubstituted iduronic acid-1-phosphate (IdoA-1-P) claim 1 , substituted or unsubstituted N-acetylglucosamine (GlcNAc) claim 1 , substituted or unsubstituted N-acetylglucosamine-1-phosphate (GlcNAc-1-P) claim 1 , substituted or unsubstituted glucosamine (GlcNH) claim 1 , substituted or unsubstituted glucosamine-1-phosphate (GlcNH-1-P) claim 1 , substituted or unsubstituted galactose (Gal) claim 1 , substituted or unsubstituted galactose-1-phosphate (Gal-1-P) claim 1 , substituted or unsubstituted galacturonic acid (GalA) claim 1 , substituted or unsubstituted galacturonic acid-1-phosphate (GalA-1-P) claim 1 , substituted or unsubstituted N-acetylgalactosamine (GalNAc) claim 1 , substituted or unsubstituted N-acetylgalactosamine-1-phosphate (GalNAc-1-P) claim 1 , substituted or unsubstituted galactosamine (GalNH) claim 1 , substituted or unsubstituted galactosamine-1-phosphate (GalNH-1-P) claim 1 , substituted or unsubstituted mannose (Man) claim 1 , substituted or unsubstituted mannose-1-phosphate (Man-1-P) ...

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 ...

COLLOIDAL DISPERSIONS OF POLY ALPHA-1,3-GLUCAN BASED POLYMERS

A colloidal dispersion is disclosed comprising poly alpha-1,3-glucan or poly alpha-1,3-1,6-glucan and a solvent. The colloidal dispersion has utility in various applications, including food, oil field, pharmaceutical, personal care and specialty industries. 1. A colloidal dispersion comprising:(a) poly alpha-1,3-glucan or poly alpha-1,3-1,6-glucan; and(b) a solvent.2. The colloidal dispersion of claim 1 , wherein the poly alpha-1 claim 1 ,3-1 claim 1 ,6-glucan has alpha-1 claim 1 ,3 and alpha-1 claim 1 ,6-glycosidic linkages comprising:(a) at least 30% of the glycosidic linkages are alpha-1,3 linkages; and(b) at least 30% of the glycosidic linkages are alpha-1,6 linkages.3. The colloidal dispersion of claim 1 , wherein the poly alpha-1 claim 1 ,3-glucan and the poly alpha-1 claim 1 ,3-1 claim 1 ,6-glucan comprise particles with an average particle diameter size of between 5 nm and 200 nm.4. The colloidal dispersion of claim 3 , wherein the particles have a spherical or cylindrical shape.5. The colloidal dispersion of claim 3 , wherein the particles form aggregates with an average aggregate diameter size of between 10 nm and 200 μm.6. The colloidal dispersion of claim 1 , wherein the solvent is water.7. The colloidal dispersion of claim 1 , wherein the poly alpha-1 claim 1 ,3-glucan claim 1 , or poly alpha-1 claim 1 ,3-1 claim 1 ,6-glucan constitutes between 0.1% by wt. and 15% by wt. of the total colloidal dispersion.8. The colloidal dispersion of claim 1 , wherein the colloidal dispersion has a viscosity of at least 10 cPs.9. The colloidal dispersion of claim 1 , wherein the colloidal dispersion has a pH between 1 and 14.10. The colloidal dispersion of claim 1 , wherein the colloidal dispersion has a viscosity and a pH wherein the viscosity changes less than 10% while the pH is changed over a range between 2 and 11.11. The colloidal dispersion of claim 1 , wherein the colloidal dispersion further comprises a salt or surfactant wherein the colloidal dispersion has a ...

Synthesis of new sialooligosaccharide derivatives

The invention relates to a method for the synthesis of compounds of general formula (1A) and salts thereof wherein one of the R groups is an α-sialyl moiety and the other is H, X 1 represents a carbohydrate linker, A is a D -glucopyranosyl unit optionally substituted with fucosyl, R 1 is a protecting group that is removable by hydrogenolysis, the integer in is 0 or 1, by a transsialidation reaction.

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 ...

SYNTHESIS AND USE OF ISOTOPICALLY-LABELLED GLYCANS

Isotopically-labelled glycans and their synthesis and use as internal standards in the analysis by mass spectrometry of glycan mixtures is described. The methods of synthesis described herein may be used conveniently to prepare libraries of heavy glycans for use in the qualitative and quantitative identification of glycans in natural samples. 1. A method for the synthesis of an isotopically-labelled glycan for use as a mass spectrometry internal standard , the method comprising:acylating an oligosaccharide core structure with an isotopically-labelled acylating agent, wherein the oligosaccharide core structure is optionally protected with one or more protecting groups, to obtain an isotopically-labelled oligosaccharide core structure; andenzymatically derivatising the resultant isotopically-labelled oligosaccharide to obtain the isotopically-labelled glycan.2. The method of claim 1 , wherein the enzymatic derivatisation comprises an enzymatic hydrolysis step to remove a terminal sugar unit.3. The method of claim 1 , wherein the enzymatic derivatisation comprises an enzymatic elongation step with a glycosyltransferase and a suitable sugar donor claim 1 , optionally wherein the enzymatic elongation step incorporates a sugar unit that is itself isotopically-labelled.4. The method of claim 1 , wherein the isotopically-labelled oligosaccharide core structure is protected with one or more protecting groups during enzymatic derivatisation.5. The method of claim 4 , wherein the isotopically-labelled oligosaccharide core structure is protected with one or more optionally substituted benzyl groups.6. The method of claim 1 , wherein the oligosaccharide core structure comprises a disaccharide motif claim 1 , the disaccharide motif comprising a first monosaccharide unit and a second monosaccharide unit claim 1 , wherein at least one of the first monosaccharide unit and/or second monosaccharide unit comprises an amino group and acylation occurs at the amino group(s).12. The method ...

BIOSYNTHESIS PRODUCTION OF STEVIOL GLYCOSIDES AND PROCESSES THEREFORE

The present invention relates to the production of steviol glycoside rebaudiosides D4, WB1 and WB2 and the production of rebaudioside M from Reb D4. 154.-. (canceled)56Pichia pastoris. The reaction mixture of claim 55 , wherein the beta-glucosidase is a beta-glucosidase.57. The reaction mixture of claim 55 , wherein the beta-glucosidase comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 5.58. The reaction mixture of claim 55 , wherein the beta-glucosidase comprises the amino acid sequence of SEQ ID NO: 5.59. The reaction mixture of claim 55 , wherein the polynucleotide comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 6.60. The reaction mixture of claim 55 , wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 6.61. The reaction mixture of claim 55 , wherein the cell is a yeast cell.62. The reaction mixture of claim 55 , wherein the cell is a bacterial cell.63. The reaction mixture of claim 55 , wherein the cell is a plant cell.65. The reaction mixture of claim 64 , wherein the UDP glycosyltransferase comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 9.66. The reaction mixture of claim 64 , wherein the UDP glycosyltransferase comprises the amino acid sequence of SEQ ID NO: 9.67. The reaction mixture of claim 64 , wherein the polynucleotide comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 10.68. The reaction mixture of wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO: 10.69. The reaction mixture of claim 64 , wherein the cell is a yeast cell.70. The reaction mixture of claim 64 , wherein the cell is a bacterial cell.71. The reaction mixture of claim 64 , wherein the cell is a plant cell.72. The reaction mixture of claim 64 , further comprising a substrate selected from the group consisting of: sucrose claim ...

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.

GLUCOSYL STEVIA COMPOSITION

Glucosyl compositions are prepared from steviol glycosides of Bertoni. The glucosylation was performed by cyclodextrin glucanotransferase using the starch as source of glucose residues. The compositions mainly comprise glucosyl derivatives with superior taste characteristics and can be used as sweetness enhancers, flavor enhancers and sweeteners in foods, beverages, cosmetics and pharmaceuticals. 113-. (canceled)14SteviaStevia rebaudiana. The food claim 17 , beverage claim 17 , cosmetic or pharmaceutical product of claim 17 , further comprising an additional sweetening agent selected from the group consisting of: extract claim 17 , steviol glycosides claim 17 , stevioside claim 17 , rebaudioside A claim 17 , rebaudioside B claim 17 , rebaudioside C claim 17 , rebaudioside D claim 17 , rebaudioside E claim 17 , rebaudioside F claim 17 , dulcoside A claim 17 , steviolbioside claim 17 , rubusoside claim 17 , other steviol glycosides found in Bertoni plant and mixtures thereof claim 17 , Luo Han Guo extract claim 17 , mogrosides claim 17 , high-fructose corn syrup claim 17 , corn syrup claim 17 , invert sugar claim 17 , fructooligosaccharides claim 17 , inulin claim 17 , inulooligosaccharides claim 17 , coupling sugar claim 17 , maltooligosaccharides claim 17 , maltodextins claim 17 , corn syrup solids claim 17 , glucose claim 17 , maltose claim 17 , sucrose claim 17 , lactose claim 17 , aspartame claim 17 , saccharin claim 17 , sucralose claim 17 , sugar alcohols claim 17 , and a combination thereof.15. The food claim 17 , beverage claim 17 , cosmetic or pharmaceutical product of claim 17 , further comprising an additional flavoring agent selected from the group consisting of: lemon claim 17 , orange claim 17 , fruity claim 17 , banana claim 17 , grape claim 17 , pear claim 17 , pineapple claim 17 , mango claim 17 , bitter almond claim 17 , cola claim 17 , cinnamon claim 17 , sugar claim 17 , cotton candy claim 17 , vanilla claim 17 , and a combination thereof.16. The ...

Glucosylated steviol glycoside composition as a flavor modifier

A taste and flavor profile enhancing composition is described. The composition includes glucosylated steviol glycosides which can enhance the intensity of a taste and/or a flavor in a food or beverage product.

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 ...

USE OF OCTAKETIDE SYNTHASES TO PRODUCE KERMESIC ACID AND FLAVOKERMESIC ACID

A method for producing an octaketide derived aromatic compound of interest (e.g. carminic acid), wherein the method comprises (I): heterologous expression of a recombinantly introduced Type III polyketide synthase (PKS) gene encoding an octaketide synthase (OKS) to obtain non-reduced octaketide in vivo within the recombinant host cell and (II): converting in vivo the non-reduced octaketide of step (I) into a C-Caromatic compound of interest (e.g. carminic acid). 1. (canceled)2. A method for producing an octaketide derived aromatic compound , wherein the method comprises:(I): contacting in vivo in a recombinant host cell comprising a recombinantly introduced Type III polyketide synthase (PKS) gene encoding an octaketide synthase (OKS), wherein the OKS is of a different genus than the host cell:(i): a starter unit and an extender unit with said OKS such that the starter and extender units convert into a non-reduced octaketide;{'sub': 14', '34, '(II): converting in vivo within the recombinant host cell the non-reduced octaketide of step (I) into a C-Caromatic aglycon compound via at least one in trans acting aromatase/cyclase, wherein the aromatic aglycon compound is not SEK4 and/or SEK4B;'}{'sub': 14', '34, '(III): the recombinant host cell further comprises a glycosyltransferase gene encoding a glycosyltransferase having at least 90% sequence identity to SEQ ID NO: 2 or amino acids 1 to 468 of SEQ ID NO:2, which glycosylates the aromatic aglycon compound produced in step (II) into a C-Caromatic glycoside compound; and'}(IV): isolating the aromatic glycoside compound of step (III) so as to get a composition, wherein the composition comprises less than 1% w/w dry matter of recombinant host cell material, and wherein the recombinant host cell is a yeast cell.3. The method of claim 2 , wherein the recombinant host cell is a growing recombinant host cell and step (I) and step (II) comprise:(I): contacting in vivo in a growing recombinant host cell comprising a ...

NEW CLASS OF SUCROSE ESTERS AND A METHOD FOR THEIR PREPARATION

The present invention relates to a new class of sucrose esters and a method for their preparation. 1B. megaterium. A method for the preparation of a P-D-fructofuranosyl-(2 ,1)-α-D-uronic acid or an ester thereof which comprises the step of fructosvlating a D-uronic acid salt or ester thereof in the presence of levansucrase (Bm-Ls).2. The method claim 1 , wherein the D-uronic acid is D-galacturonic acid or D-glucuronic acid.3. The method of claim 1 , wherein the D-uronic acid salt is an alkali metal salt.4. The method of claim 1 , residue having 1 to 30 carbon atoms which may be interrupted by one or more heteroatoms and which may be substituted by one or more functional groups.5. The method of claim 1 , wherein the method is for the preparation of a β-D-fructofuranosyl-(2 claim 1 ,1)-α-D-uronic acid ester claim 1 , further wherein an esterificaiton step is conducted prior or after the fructosylation step.6. The method of claim 5 , wherein the esterification step is conducted in the presence of tetrabutylammonium fluoride.7. A method for the preparation of a D-uronic acid ester comprising the step of reacting D-uronic acid with an organic halide in the presence of tetrabutylammonium fluoride.8. The method of claim 7 , wherein the organic halide has the formula Hal-R claim 7 , wherein R is a hydrocarbon residue having 1 to 30 carbon atoms which may be interrupted by one or more heteroatoms and which may be substituted by one or more functional groups.9. The method of claim 7 , wherein the halide is I.10. β-D-fructofuranosyl-(2 claim 7 ,1)-α-D-uronic acid mono ester.11. β-D-fructofuranosyl-(2 claim 7 ,1)-α-D-galacturonic acid.12. β-D-fructofuranosyl-(2 claim 7 ,1)-α-D-6-p-toluensulfonylglycopyranoside.13. β-D-fructofuranosyl-(2 claim 7 ,1)-α-D-6-azidoglycopyranoside.14. The method of claim 3 , wherein the alkali metal salt is sodium salt. The present invention relates to a new class of sucrose esters and a method for their preparation.Carbohydrates are an important ...

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.

Mapping Cytosine Modifications

Methods, compositions and kits for selectively altering and detecting modified cytosine residues are provided.

ENZYMATIC HYDROLYSIS OF DISACCHARIDES AND OLIGOSACCHARIDES USING ALPHA-GLUCOSIDASE ENZYMES

A method is disclosed for hydrolyzing an alpha-1,3 or alpha-1,6 glucosyl-glucose linkage in a saccharide (disaccharide or oligosaccharide). This method comprises contacting the saccharide with an alpha-glucosidase enzyme such as transglucosidase under suitable conditions, during which contacting step the enzyme hydrolyzes at least one alpha-1,3 or alpha-1,6 glucosyl-glucose linkage of the saccharide. This method is useful for reducing the amount of oligosaccharides in a filtrate isolated from a glucan synthesis reaction, for example. 115-. (canceled)16. A method that comprises:(a) providing an alpha-glucan synthesis reaction that comprises a soluble disaccharide or oligosaccharide byproduct, wherein at least 90% of the glycosidic linkages of the alpha-glucan are alpha-1,3-glycosidic linkages; and(b) contacting the soluble disaccharide or oligosaccharide byproduct with an alpha-glucosidase enzyme, wherein the amount of the soluble disaccharide or oligosaccharide byproduct is reduced compared to the amount of the soluble disaccharide or oligosaccharide byproduct that was present prior to said contacting.17. The method of claim 16 , wherein the alpha-glucosidase enzyme is immobilized.18. The method of claim 16 , wherein the degree of polymerization of the oligosaccharide byproduct prior to said contacting with the alpha-glucosidase is 3 to 7.19. The method of claim 16 , wherein said contacting of the soluble disaccharide or oligosaccharide byproduct with the alpha-glucosidase enzyme is performed in the alpha-glucan synthesis reaction.20. The method of claim 16 , further comprising obtaining a fraction of the alpha-glucan synthesis reaction that comprises the soluble disaccharide or oligosaccharide byproduct claim 16 , wherein said contacting of the soluble disaccharide or oligosaccharide byproduct with the alpha-glucosidase enzyme is performed in the fraction.21. The method of claim 20 , wherein the fraction is a filtrate of the alpha-glucan synthesis reaction.22. The ...

Production of Defined Monodisperse Heparosan Polymers and Unnatural Polymers with Polysaccharide Synthases

A methodology for polymer grafting by a polysaccharide synthase allows the creation of a variety of glycosaminoglycan oligosaccharides that may have a natural, chimeric, hybrid, and/or unnatural sugar structure and/or a targeted size (i.e., substantially monodisperse in size).

ENGINEERED GLYCOSYLTRANSFERASES AND STEVIOL GLYCOSIDE GLUCOSYLATION METHODS

The present invention provides engineered glycosyltransferase (GT) enzymes, polypeptides having GT activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. The present invention provides engineered sucrose synthase (SuS) enzymes, polypeptides having SuS activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. The present invention also provides compositions comprising the GT enzymes and methods of using the engineered GT enzymes to make products with β-glucose linkages. The present invention further provides compositions and methods for the production of rebaudiosides (e.g., rebaudioside M, rebaudioside A, rebaudioside I, and rebaudioside D). The present invention also provides compositions comprising the SuS enzymes and methods of using them. Methods for producing GT and SuS enzymes are also provided. 1. An engineered glycosyltransferase comprising a polypeptide sequence that is at least 60% , 65% , 70% , 75% , 80% , 85% , 86% , 87% , 88% , 89% , 90% , 91% , 92% , 93% , 94% , 95% , 96% , 97% , 98% , 99% , or more sequence identity to SEQ ID NO: 758 , 770 , 792 , 954 , 1002 , 1054 , 2600 , 2718 , 2814 , 2884 , 3016 , 3082 , 3244 , 3502 , 3346 , 3502 , 3696 , 3956 , 4256 , 4550 , 7324 , and 7784.2. The engineered glycosyltransferase of claim 1 , wherein said polypeptide sequence of said engineered glycosyltransferase comprises at least one mutation or mutation set at one or more positions selected from 69/173/175/243/246/354/365/383/399 claim 1 , 69/173/243/383/399 claim 1 , 56/191/354/383/399 claim 1 , 70/225/246/409/413 claim 1 , 70/115/225/409 claim 1 , 70/225/413 claim 1 , 70/225/247 claim 1 , 74/310/396/424 claim 1 , 74/396 claim 1 , and 173/175/191/365/383/399 claim 1 , wherein said positions are numbered with reference to SEQ ID NO:758.3. The engineered glycosyltransferase of claim 1 , wherein said ...

ENGINEERED GLYCOSYLTRANSFERASES AND STEVIOL GLYCOSIDE GLUCOSYLATION METHODS

The present invention provides engineered glycosyltransferase (GT) enzymes, polypeptides having GT activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. The present invention provides engineered sucrose synthase (SuS) enzymes, polypeptides having SuS activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. The present invention also provides compositions comprising the GT enzymes and methods of using the engineered GT enzymes to make products with β-glucose linkages. The present invention further provides compositions and methods for the production of rebaudiosides (e.g., rebaudioside M, rebaudioside A, rebaudioside I, and rebaudioside D). The present invention also provides compositions comprising the SuS enzymes and methods of using them. Methods for producing GT and SuS enzymes are also provided. 1. An engineered sucrose synthase comprising a polypeptide sequence that is at least 60% , 65% , 70% , 75% , 80% , 85% , 86% , 87% , 88% , 89% , 90% , 91% , 92% , 93% , 94% , 95% , 96% , 97% , 98% , 99% , or more sequence identity to SEQ ID NO:72 , 74 , 1080 , 1158 , 1222 , 1392 , 1456 , 1582 , 1764 , 1804 , 1840 , 2064 , 2432 , 2510 , 7506 , and/or 8420.2. The engineered sucrose synthase of claim 1 , wherein said polypeptide sequence of said engineered sucrose synthase comprises at least one mutation or mutation set at one or more positions selected from 4/9/349/532 claim 1 , 4/13/113/343/532 claim 1 , 4/13/113/532 claim 1 , 4/33/47/52/343/532 claim 1 , 4/47/52/532 claim 1 , 4/113/532 claim 1 , 4/13/113 claim 1 , 4/13/532 claim 1 , 4/33/113 claim 1 , 4/343 claim 1 , 7 claim 1 , 8 claim 1 , 44 claim 1 , 95 claim 1 , 117/440 claim 1 , 136 claim 1 , 221 claim 1 , 343/532 claim 1 , 440 claim 1 , 444 claim 1 , 478 claim 1 , 532 claim 1 , 583 claim 1 , 611 claim 1 , 615 claim 1 , 615/789 claim 1 , 695 claim 1 , ...

HIGH-PURITY STEVIOL GLYCOSIDES

Methods of preparing highly purified steviol glycosides, particularly rebaudiosides A, D and X are described. The method includes expression of UDP-glucosyltransferases from Bertoni, which are capable converting certain steviol glycosides to rebaudiosides A, D and X. The highly purified rebaudiosides A, D and X, are useful as non-caloric sweetener in edible and chewable compositions such as any beverages, confectioneries, bakery products, cookies, and chewing gums. 126.-. (canceled)27. A method for adding at least one glucose unit to a steviol glycoside substrate comprising rebaudioside E to provide a target steviol glycoside , comprising contacting the steviol glycoside substrate with a biocatalyst protein enzyme comprising UDP-glucosyltransferase , wherein the target steviol glycoside is Rebaudioside X.28. The method of claim 27 , wherein the biocatalyst protein enzyme is a recombinant protein.29. The method of claim 27 , further comprising the step of purifying the Rebaudioside X to a purity of greater than about 80% by weight.30. The method of claim 27 , further comprising the step of purifying the Rebaudioside X to a purity of greater than about 90% by weight.31. The method of claim 27 , further comprising the step of purifying the Rebaudioside X to a purity of greater than about 95% by weight.32. The method of claim 27 , wherein the UDP-glucosyltransferase is expressed in a host microorganism.33Escherichia coli, Saccharomyces species, AspergillusPischia. The method of claim 32 , wherein the host microorganism is selected from the group consisting of: species claim 32 , and species. This application is a continuation of U.S. patent application Ser. No. 16/694,893, filed Nov. 25, 2019, which is a continuation of U.S. patent application Ser. No. 15/400,325, filed Jan. 6, 2017, now U.S. Pat. No. 10,485,257, which is a continuation of U.S. patent application Ser. No. 14/954,213, now abandoned, which is a divisional of U.S. patent application Ser. No. 14/469,076, ...

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 ...

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. 121.-. (canceled)22. A method for producing an N-acetylglucosamine-containing oligosaccharide in a bacterium comprising: 'culturing said bacterium in the presence of lactose.', 'providing a bacterium, said bacterium comprising an exogenous UDP-GlcNAc:Galα/β-R 3-N-acetylglucosaminyltransferase gene and a functional lactose permease gene; and'}23. The method of claim 22 , wherein said bacterium comprises an increased UDP-GlcNAc production capability.24. The method of claim 22 , wherein said increased UDP-GlcNAc production capability comprises overexpression of a nagC gene claim 22 , a glmS gene claim 22 , a glmY gene claim 22 , a glmZ gene or any combination thereof.25E. coli. The method of claim 22 , wherein said increased UDP-GlcNAc production capability comprises overexpression of nagC gene.26. The method of claim 22 , wherein said increased UDP-GlcNAc production capability comprises overexpression of nagC and glmS.27. The method of claim 22 , wherein said increased UDP-GlcNAc production capability comprises overexpression of nagC and glmY.28. The method of claim 22 , wherein said increased UDP-GlcNAc production capability comprises overexpression of nagC and glmZ.29. The method of claim 22 , wherein said N-acetylglucosamine-containing oligosaccharide comprises any one selected from Lacto-N-triose 2 (LNT2) claim 22 , Lacto-N-tetraose (LNT) claim 22 , Lacto-N-neotetraose (LNnT) claim 22 , Lacto-N-fucopentaose I (LNF I) claim 22 , Lacto-N-fucopentaose II (LNF II) claim 22 , Lacto-N-fucopentaose III (LNF III) claim 22 , Lacto-N-fucopentaose V (LNF V) claim 22 , Lacto-N-difucohexaose I (LDFH I) claim 22 , Lacto-N-difucohexaose II (LDFH II) claim 22 , and Lacto-N-neodifucohexaose II (LFNnDFH II).30E. coli.. The method of claim 22 , wherein said bacterium is31. ...

Process for the purification of a neutral human milk oligosaccharide (hmo) from microbial fermentation

The invention relates to a process for the purification of a neutral human milk oligosaccharide (HMO) from a fermentation broth, the process comprising the steps of: (i) separating biomass from the fermentation broth to provide a cmde solution; (ii) treating the crude solution with: (a) a cation-exchange material; (b) an anion-exchange material; and (c) a cation-exchange adsorbent resin; thereby obtaining a purified solution containing the neutral human milk oligosaccharide. Further, the invention relates to a process for fermentatively producing HMO in a fermentation broth and purifying the HMO from the broth.

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 PRODUCING FLAVONOID INCLUSION COMPOUND

A method for producing a flavonoid inclusion compound, including a cleaving step including treating a sparingly soluble flavonoid having a rhamnoside structure with an enzyme having a rhamnosidase activity in the presence of a cyclodextrin to cleave a rhamnose. According to the production method of the present invention, a flavonoid inclusion compound and a composition of flavonoid glycosides having excellent solubility in water can be efficiently produced, so that the compound and composition can be suitably utilized in the fields of medicaments, foodstuff, health foods, foods for specified health use, cosmetics, and the like. 1. A flavonoid inclusion compound-containing composition comprising a flavonoid inclusion compound and a rhamnose , wherein the flavonoid inclusion compound is obtained by a production method , wherein the method comprises an elimination step comprising treating a sparingly soluble flavonoid having a rhamnoside structure with an enzyme having a rhamnosidase activity in the presence of a cyclodextrin to eliminate a rhamnose , wherein a molar ratio of a flavonoid in the flavonoid inclusion compound and the rhamnose (rhamnose/flavonoid) is from 0.8 to 1.2 , and wherein the cyclodextrin is one or more members selected from the group consisting of β-cyclodextrin , branched β-cyclodextrin , and γ-cyclodextrin.2. The flavonoid inclusion compound-containing composition according to claim 1 , wherein the sparingly soluble flavonoid having a rhamnoside structure is one or more members selected from the group consisting of rutin claim 1 , hesperidin claim 1 , narirutin claim 1 , naringin claim 1 , diosmin claim 1 , eriocitrin claim 1 , myricitrin claim 1 , neohesperidin claim 1 , luteolin-7-rutinoside claim 1 , delphinidin-3-rutinoside claim 1 , cyanidin-3-rutinoside claim 1 , isorhamnetin-3-rutinoside claim 1 , kaempferol-3-rutinoside claim 1 , apigenin-7-rutinoside claim 1 , and acacetin-7-rutinoside.3. The flavonoid inclusion compound-containing ...

Increasing activity of 2'fucosyllactose transporters endogenous to microbial cells

A fermentation broth which includes a microbial cell which has been subjected to a condition under which the activity an endogenous transporter for 2′ fucosyllactose is increased. Also provided are methods for increasing export of 2′ fucosyllactose from a microbial cell, methods for identifying an endogenous yeast transporter of 2′ fucosyllactose, and microbial cells genetically engineered to increase the activity of an endogenous transporter of 2′ fucosyllactose. 1. A fermentation broth comprising a microbial cell having increased export of 2′ fucosyllactose , the microbial cell having been subjected to a condition under which the activity of an endogenous transporter is increased relative to the activity of the endogenous transporter in the microbial cell in the absence of subjecting the microbial cell to the condition.2. The fermentation broth of wherein the condition comprises one or more of growing the microbial cell in a medium comprising amino acids claim 1 , growing the microbial cell in a glucose limited medium claim 1 , and growing the microbial cell in a medium comprising ethanol.3. The fermentation broth of wherein the microbial cell is a yeast cell.4. A method for increasing the export of 2′ fucosyllactose from a microbial cell comprising:a) obtaining a 2′FL-containing microbial cell; andb) subjecting the microbial cell to a condition under which the activity of an endogenous transporter is increased relative to the activity of the endogenous transporter in the microbial cell in the absence of subjecting the microbial cell to the condition.5. The method of wherein subjecting the microbial cell to a condition under which the activity of an endogenous transporter is increased comprises one or more of growing the microbial cell in a medium comprising amino acids claim 4 , growing the microbial cell in a glucose limited medium claim 4 , and growing the microbial cell in a medium comprising ethanol.6. The method of wherein the microbial cell is a yeast cell. ...

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 % ...

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 ...

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 ...

PROCESS FOR PRODUCING A PARTICULATE COMPOSITION COMPRISING CRYSTALLINE ALPHA, ALPHA-TREHALOSE DI-HYDRATE

A process for enabling the production of a particulate composition containing crystalline trehalose dihydrate is provided. Including allowing an α-glycosyltrehalose-forming enzyme to act on liquefied starch derived from a microorganism of the genus and a trehalose-releasing enzyme derived from a microorganism of the genus along with a starch debranching enzyme and a cyclomaltodextrin glucanotransferase; allowing glucoamylase to act on the resulting mixture to obtain a saccharide solution containing α,α-trehalose; precipitating crystalline α,α-trehalose dihydrate from the above saccharide solution; collecting the precipitated crystalline α,α-trehalose dihydrate by a centrifuge; and ageing and drying the collected crystals. Cyclomaltodextrin glucanotransferase derived from a microorganism of the genus or a mutant enzyme thereof is used to increase the α,α-trehalose content in the saccharide solution to over 86.0% by weight, on a dry solid basis, without passing through a fractionation step by column chromatography. 15-. (canceled)6. A process for producing a particulate composition comprising crystalline α-trehalose dihydrate with an α ,α-trehalose content of 98.0% by weight or more , on a dry solid basis , said process comprising the steps of:allowing to act on liquefied starch an α-glycosyltrehalose-forming enzyme and a trehalose-releasing enzyme along with a starch debranching enzyme and a cyclomaltodectrin glucanotransferase;allowing a glucoamylase to act on the resulting mixture to obtain a saccharide solution comprising α,α-trehalose;precipitating crystalline α,α-trehalose dihydrate from said saccharide solution by a controlled cooling method or semi-controlled cooling method;collecting the precipitated crystalline α,α-trehalose dihydrate by a centrifuge; andageing and drying the collected crystals to obtain a particulate composition comprising crystalline α,α-trehalose dihydrate,wherein said cyclomaltodextrin glucanotransferase is used to increase the α,α- ...

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.

Biosynthetic production of steviol glycosides and processes therefore

The present invention relates to the production of steviol glycoside rebaudiosides D4, WB1 and WB2 and the production of rebaudioside M from Reb D4.

Biosynthesis Of Human Milk Oligosaccharides In Engineered Bacteria

The invention provides compositions and methods for engineering bacteria 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 a bacterium, said bacterium comprising a functional β-galactosidase gene, an exogenous fucosyltransferase gene, a GDP-fucose synthesis pathway, a functional lactose permease gene;culturing said bacterium in the presence of lactose; andretrieving a fucosylated oligosaccharide from said bacterium or from a culture supernatant of said bacterium.2E. coli. The method of claim 1 , wherein said β-galactosidase gene comprises an lacZ gene.3. The method of claim 1 , wherein said β-galactosidase gene is an endogenous β-galactosidase gene or an exogenous β-galactosidase gene.4. The method of claim 1 , wherein said bacterium accumulates an increased intracellular lactose pool claim 1 , and produces a low level of β-galactosidase.5. The method of claim 1 , wherein said exogenous fucosyltransferase gene encodes α(1 claim 1 ,2) fucosyltransferase or α(1 claim 1 ,3) fucosyltransferase.6Bacteroides fragilis. The method of claim 5 , wherein said α(1 claim 5 ,2) fucosyltransferase gene comprises a wcfW gene.7Helicobacter pylori. The method of claim 5 , wherein said α(1 claim 5 ,3) fucosyltransferase gene comprises a 26695 futA gene.8. The method of claim 5 , wherein said bacterium comprises both an exogenous fucosyltransferase gene encoding α(1 claim 5 ,2) fucosyltransferase and an exogenous fucosyltransferase gene encoding α(1 claim 5 ,3) fucosyltransferase.9. The method of claim 1 , wherein said GDP-fucose synthesis pathway comprises endogenous enzymes or exogenous enzymes.10. The method of claim 1 , wherein said lactose permease gene is an endogenous lactose permease gene or an exogenous lactose permease gene.11. A method for producing a fucosylated oligosaccharide in a bacterium claim 1 , comprisingproviding an ...

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 ...

BOVINE MILK OLIGOSACCHARIDES

Oligosaccharides from bovine milk, whey and dairy products, and methods of producing bovine milk oligosaccharides are provided. 1BifidobacteriuminfantisB. breve. A method of treating gastrointestinal tract microbiota imbalances in an infant , wherein the method comprises administering to the infant a sufficient amount of a composition of subsp. or in or in conjunction with an infant formula ,wherein the infant formula comprises one or more of the following fucosylated oligosaccharides as found in bovine milk:an oligosaccharide consisting of 3 Hexose (Hex) moieties, 4 n-acetyl hexosamine (HexNAc) moieties and 1 fucose (Fuc) moiety;an oligosaccharide consisting of 4 Hex moieties, 4 HexNAc moieties, and 1 Fuc moiety;an oligosaccharide consisting of 3 Hex moieties, 5 HexNAc moieties, and 1 Fuc moiety;an oligosaccharide consisting of 5 Hex moieties, 4 HexNAc moieties, and 1 Fuc moiety;an oligosaccharide consisting of 4 Hex moieties, 5 HexNAc moieties, and 1 Fuc moiety; andan oligosaccharide consisting of 3 Hex moieties, 6 HexNAc moieties, and 1 Fuc moiety;and wherein the composition stimulates the production of a bifidobacterial secretion that reduces colonization of enteropathogenic bacteria in the gut of the infant; modulates signals generated by enteroendocrine and gut epithelial cells in the infant; improves at least one biomarker of gut health in the infant; and/or increases gut colonization and persistence of probiotic bacteria in the infant gut.2. The method of wherein the infant formula further comprises one or more oligosaccharides comprising 2 hexose (Hex) moieties and 1 fucose (Fuc) moiety; 4 Hex moieties and 1 Fuc moiety; 2 HEX moieties claim 1 , 1 n-acetyl hexosamine (HexNAc) moiety and 1 Fuc moiety; 3 Hex moieties and 1 HexNAc moiety; 3 Hex moieties and 6 HexNAc moieties; 4 Hex moieties and 3 HexNAc moieties; 3 Hex moieties and 4 HexNAc moieties; 6 Hex moieties and 2 HexNAc moieties; 4 Hex moieties and 4 HexNAc moieties; 3 Hex moieties and 5 HexNAc moieties ...

Method for producing 3'-fucosyllactose using corynebacterium glutamicum

Disclosed is a method for producing 3′-fucosyllactose using a wild Corynebacterium glutamicum strain. In addition, using the Corynebacterium glutamicum strain, which is a GRAS strain, 3′-fucosyllactose can be produced at a high concentration, high yield and high productivity.

BIOSYNTHESIS OF HUMAN MILK OLIGOSACCHARIDES IN ENGINEERED BACTERIA

The invention provides compositions and methods for engineering bacteria 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 a bacterium, said bacterium comprising a functional β-galactosidase gene, an exogenous fucosyltransferase gene, a GDP-fucose synthesis pathway, a functional lactose permease gene;culturing said bacterium in the presence of lactose; andretrieving a fucosylated oligosaccharide from said bacterium or from a culture supernatant of said bacterium.2E. coli. The method of claim 1 , wherein said β-galactosidase gene comprises an lacZ gene.3. The method of claim 1 , wherein said β-galactosidase gene is an endogenous β-galactosidase gene or an exogenous β-galactosidase gene.4. The method of claim 1 , wherein said bacterium accumulates an increased intracellular lactose pool claim 1 , and produces a low level of β-galactosidase.5. The method of claim 1 , wherein said exogenous fucosyltransferase gene encodes α(1 claim 1 ,2) fucosyltransferase or α(1 claim 1 ,3) fucosyltransferase.6Bacteroides fragilis. The method of claim 5 , wherein said α(1 claim 5 ,2) fucosyltransferase gene comprises a wcfW gene.7Helicobacter pylori. The method of claim 5 , wherein said α(1 claim 5 ,3) fucosyltransferase gene comprises a 26695 futA gene.8. The method of claim 5 , wherein said bacterium comprises both an exogenous fucosyltransferase gene encoding α(1 claim 5 ,2) fucosyltransferase and an exogenous fucosyltransferase gene encoding α(1 claim 5 ,3) fucosyltransferase.9. The method of claim 1 , wherein said GDP-fucose synthesis pathway comprises endogenous enzymes or exogenous enzymes.10. The method of claim 1 , wherein said lactose permease gene is an endogenous lactose permease gene or an exogenous lactose permease gene.11. A method for producing a fucosylated oligosaccharide in a bacterium claim 1 , comprisingproviding an ...

STEVIA SWEETENER AND MANUFACTURING PROCESS

A method of preparing a glucosylated steviol glycoside composition involving by treating stevia materials with cyclodextrin glycosyltransferase. 1Stevia. A process for producing a highly purified glucosyl composition , comprising the steps of:{'i': 'Stevia rebaudiana', '(a) providing steviol glycosides extracted from in an acidic solution, thereby forming an acidic mixture;'}(b) contacting the acidic mixture with a first amount of cyclodextrin glycosyltransferase to form a first enzyme treated mixture;(c) maintaining the first enzyme treated mixture at a temperature of about 65° C. for between 4 and 10 hours;(d) contacting the first enzyme treated mixture with a second amount of cyclodextrin glycosyltransferase to form a second enzyme treated mixture;(e) maintaining the second enzyme treated mixture at a temperature of about 65° C. for between 6 and 10 hours;(f) inactivating the cyclodextrin glycosyltransferase in the second enzyme treated mixture by raising the temperature of the second enzyme treated mixture to between about 90° C. and about 100° C., thereby forming an enzyme inactivated mixture; and{'i': 'Stevia', '(g) purifying the enzyme inactivated mixture to form the glucosyl composition.'}2. The process of claim 1 , wherein prior to step (a) the process comprises the steps of:{'i': 'Stevia rebaudiana', '(i) soaking materials in water to extract steviol glycosides, thereby forming an aqueous mixture;'}(ii) passing the aqueous mixture over a column containing activated charcoal, thereby adsorbing steviol glycosides on the activated charcoal; and(iii) eluting the adsorbed steviol glycosides from the activated charcoal.3. The process of claim 1 , wherein the acidic mixture of step (a) is at a temperature of about 65° C.4. The process of claim 1 , wherein the acidic mixture comprises about 15 weight percent steviol glycosides.5. The process of claim 1 , wherein the acidic mixture comprises about 15 weight percent maltodextrin.6. The process of claim 1 , wherein ...

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 ...

METHOD FOR PRODUCING TREHALOSE EMPLOYING A TREHALOSE PHOSPHORYLASE VARIANT

The present invention relates to a method for producing trehalose, comprising the steps of mixing and reacting, in any order, (i) at least one alpha-phosphorylase capable of catalyzing the production of alpha-D-glucose 1-phosphate intermediate from a saccharide raw material, and from at least one phosphorus source; (ii) at least one trehalose phosphorylase capable of catalyzing the production of trehalose from an alpha-D-glucose 1-phosphate intermediate and a glucose substrate, wherein the trehalose phosphorylase is a trehalose phosphorylase variant with an amino acid sequence which differs from the amino acid sequence of a wild type trehalose phosphorylase in at least one amino acid position, (iii) at least one saccharide raw material which produces an alpha-D-glucose 1-phosphate intermediate and a co-product by catalytic action of the alpha-phosphorylase; and (iv) at least one phosphorus source selected from the group consisting of a phosphoric acids and an inorganic salt thereof. 1. A method for producing trehalose , comprising the steps of mixing and reacting , in any order ,a) at least one alpha-phosphorylase capable of catalyzing the production of alpha-D-glucose 1-phosphate intermediate from a saccharide raw material selected from the group consisting of sucrose and starch, and from at least one phosphorus source selected from the group consisting of a phosphoric acid and an inorganic salt thereof;b) at least one trehalose phosphorylase capable of catalyzing the production of trehalose from an alpha-D-glucose 1-phosphate intermediate and a glucose substrate, wherein the trehalose phosphorylase is a trehalose phosphorylase variant with an amino acid sequence which differs from the amino acid sequence of a wild type trehalose phosphorylase in at least one amino acid position;c) at least one saccharide raw material selected from the group consisting of sucrose and starch which produces an alpha-D-glucose 1-phosphate intermediate and a co-product selected from ...

SIALYLTRANSFERASES AND THEIR USE IN PRODUCING SIALYLATED OLIGOSACCHARIDES

Disclosed are methods, genetically engineered cells, sialyltransferases and nucleic acid molecules encoding said sialyltransferases for producing sialylated oligosaccharides as well as the use of said sialylated oligosaccharides for providing nutritional compositions. 1. A method for producing one or more sialylated oligosaccharides , the method comprisinga) providing at least one genetically engineered cell which comprises a heterologous sialyltransferase, said heterologous sialyltransferase being capable of possessing an α-2,3-sialyltransferase activity and/or an α-2,6-sialyltransferase activity for transferring a sialic acid residue from a nucleotide-activated form as donor substrate to an acceptor molecule, wherein the acceptor molecule is selected from the group consisting of lactose, lacto-N-triose II and human milk oligosaccharides;b) cultivating the at least one cell in a fermentation broth and under conditions permissive for the production of said sialylated oligosaccharide; and optionallyc) recovering said sialylated oligosaccharide.2. The method according to claim 1 , wherein the sialylated oligosaccharide is selected from the group consisting of 3′-sialyllactose claim 1 , 6′-sialyllactose claim 1 , LST-a claim 1 , LST-b claim 1 , LST-c and DSLNT.3. The method according to claim 1 , wherein the fermentation broth contains at least one carbon source claim 1 , the at least one carbon source is optionally selected from the group consisting of glucose claim 1 , fructose claim 1 , sucrose claim 1 , glycerol claim 1 , and combinations thereof.4. The method according to claim 1 , wherein the fermentation broth contains at least one selected from the group consisting of N-acetylglucosamine claim 1 , galactose and sialic acid.5. The method according to claim 1 , wherein the at least one cell is cultivated in the absence of and/or without addition of one or more selected from the group consisting of N-acetylglucosamine claim 1 , galactose and sialic acid.6. The ...

METHOD FOR OBTAINING 1-KESTOSE

The present invention discloses an industrial scale method to obtain 1-kestose by the use of a recombinant fructosyltransferase (FTF), isolated from , expressed constitutively in a non-saccharolytic yeast. In this invention, the recombinant FTF type sucrose:sucrose 1-fructosyltransferase (1-SSTrec) is produced constitutively, stable and at high yield, both in the culture supernatant and in intact cells of the host . Hence, the invention additionally provides a method for 1-SST production at industrial scale. The recombinant enzyme is then used for mass production of short-chain fructooligosaccharides (FOS), specifically 1-kestose, from sucrose. The method of the present invention establishes conditions that allow conversion rates where the synthesized FOS constitute above 55% (w/w) of the total sugars in the reaction mixture and the 1-kestose content reaches values higher than 90% (w/w) of the total FOS fraction. 1Festuca arundinacea. A method for the production of 1-kestose on an industrial scale characterized by the conversion of sucrose into 1-kestose in a bioreactor with the use of a recombinant fructosyltransferase (FTF) from expressed constitutively in a non-saccharolitic yeast.2. The method of wherein the FTF is a sucrose:sucrose 1-fructosyltransferase (1-SST).3Pichia pastoris. The method of wherein the non-saccharolitic yeast is a strain.4Pichia pastoris. The method of wherein the strain contains multiple copies of the gene encoding the 1-SST integrated in the genome.5Pichia pastoris. The method of wherein the FTF is recovered from the supernatant and/or the cell sediment of the culture.6. The method of wherein the sucrose concentration is above 400 g/L.7. The method of wherein the FTF is produced by the recombinant yeast grown in a fermentor with discontinuous claim 1 , continuous or fed-batch operation.8. The method of wherein the carbon source used for the yeast growth is a compound selected from glycerol claim 7 , glucose and sucrose of any purity degree ...

POLY ALPHA-1,3-GLUCAN FIBRIDS AND USES THEREOF AND PROCESSES TO MAKE POLY ALPHA-1,3-GLUCAN FIBRIDS

Fibrids comprising poly alpha-1,3-glucan or surface-modified poly alpha-1,3-glucan were produced and characterized. Applications and products for using these fibrids were also developed, such as in emulsification, viscosity modification, and paper. Examples of surface-modified poly alpha-1,3-glucan fibrids include those with a positive surface charge.

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: 2.2Escherichia 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 , FutN claim 1 , WblA claim 1 , FutD claim 1 , WbgN claim 1 , or Bft3/WcfB protein.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 ...

PRODUCTION OF STEVIOL GLYCOSIDES THROUGH WHOLE CELL BIOTRANSFORMATION

In various aspects and embodiments, the invention provides microbial cells and methods for producing advanced glycosylation products from lower glycosylated intermediates. The microbial cell expresses one or more UDP-dependent glycosyl transferase enzymes in the cytoplasm, for glycosylation of the intermediates. When incubating the microbial strain with a plant extract or fraction thereof comprising the intermediates, these glycosylated intermediates are available for further glycosylation by the cell, and the advanced glycosylation products can be recovered from the media and/or microbial cells. 1. A method for producing a glycosylated product from glycoside intermediates , comprising:providing a microbial cell expressing one or more UDP-dependent glycosyl transferase enzymes (UGT enzymes) intracellularly,incubating the microbial strain with the glycoside intermediates, andrecovering the glycosylated product.2. The method of claim 1 , wherein the glycoside intermediates are provided as a plant extract or fraction thereof.3. The method of claim 1 , wherein the glycoside intermediate is a glycosylated secondary metabolite claim 1 , optionally selected from a glycosylated terpenoid claim 1 , a glycosylated flavonoid claim 1 , a glycosylated cannabinoid claim 1 , a glycosylated polyketide claim 1 , a glycosylated stilbenoid claim 1 , and a glycosylated polyphenol.4. The method of claim 3 , wherein the glycoside intermediate is a glycosylated terpenoid.5. The method of claim 4 , wherein the glycoside intermediate is steviol glycoside or mogroside.6. The method of claim 1 , wherein the glycoside intermediate has one claim 1 , two claim 1 , three claim 1 , four claim 1 , or five claim 1 , glycosyl groups.7. The method of claim 6 , wherein the glycosyl groups are independently selected from glucosyl claim 6 , galactosyl claim 6 , mannosyl claim 6 , xylosyl claim 6 , and rhamnosyl groups.8. The method of claim 6 , wherein the glycosylated product has at least five glycosyl ...

PROCESS FOR PRODUCING A PARTICULATE COMPOSITION COMPRISING AN HYDROUS CRYSTALLINE 2-O-ALPHA-D-GLUCOSYL-L-ASCORBIC ACID

The invention provides a process for enabling the production of a particulate composition containing anhydrous crystalline ascorbic acid 2-glucoside that does not significantly cake even when the production yield of ascorbic acid 2-glucoside does not reach 35% by weight. The process for producing a particulate composition containing anhydrous crystalline ascorbic acid 2-glucoside, which comprises allowing a CGTase to act on a solution containing either liquefied starch or dextrin and L-ascorbic acid and then allowing a glucoamylase to act on the resulting solution to obtain a solution with an ascorbic acid 2-glucoside production yield of at least 27%, purifying the obtained solution to increase the ascorbic acid 2-glucoside content to a level of over 86% by weight, precipitating anhydrous crystalline ascorbic acid 2-glucoside by a controlled cooling method or pseudo-controlled cooling method, collecting the precipitated anhydrous crystalline ascorbic acid 2-glucoside, and ageing and drying the collected anhydrous crystalline ascorbic acid 2-glucoside. 1which comprises 2-O-α-D-glucosyl-L-ascorbic acid in an amount of over 98.0% by weight but 99.8% by weight or lower, on a dry solid basis;which has a degree of crystallinity of 90% or higher for anhydrous crystalline 2-O-α-D-glucosyl-L-ascorbic acid, when calculated based on a profile of powder X-ray diffraction analysis of said composition;and which has a reducing power of the whole composition being less than one percent by weight.. A particulate composition comprising anhydrous crystalline 2-O-α-D-glucosyl-L-ascorbic acid, The present invention relates to a process for producing a particulate composition containing anhydrous crystalline 2-O-α-D-glucosyl-L-ascorbic acid, more particularly, to a process for producing a particulate composition, containing anhydrous crystalline 2-O-α-D-glucosyl-L-ascorbic acid that significantly, more hardly cakes compared to conventional ones.Due to its advantageous physiological ...

COMPOSITION AND METHOD FOR PRODUCING CELLULOSE

A composition that efficiently produces cellulose includes cell extracts derived from a tunicate of ascidian class or yeast expressing tunicate cellulose synthase, at least one divalent cation of calcium ion and magnesium ion, cellobiose and UDP-glucose. The composition has a pH in the range of 6.6-7.2. 1. A composition for producing cellulose , comprising:a cell extract derived from a tunicate of Ascidian or Appendicularian classes, at least one divalent cation of calcium ion and magnesium ion, cellobiose and UDP-glucose, wherein the composition has a pH in the range of 6.6-7.2.2Ciona intestinalis.. The composition of claim 1 , wherein the cell extract is derived from3Ciona intestinalis.. The composition of claim 1 , wherein the cell extract is derived from tailbud stage embryos of4. A composition for producing cellulose claim 1 , the composition comprising:a cell extract may be derived from a transformed yeast which may express a transgene encoding a protein which is involved in cellulose production, at least one divalent cation of calcium ion and magnesium ion, cellobiose and UDP-glucose, wherein the composition has a pH in the range of 6.6-7.2.5. The composition of claim 4 , wherein a cell extract may be derived from a transformed yeast which may express a protein of SEQ ID NO: 2.6. The composition of claim 1 , wherein the concentration of the divalent cation is in the range of 2-8 mM.7. The composition of claim 1 , wherein the pH of the composition is buffered by MOPS.8. The composition of claim 1 , wherein the pH of the composition is 6.8.9. The composition of claim 1 , wherein the composition further comprises a stabilizer and/or a protease inhibitor.10. The composition of claim 1 , wherein the tunicate expresses a transgene encoding a protein that is involved in cellulose production.11. A method for producing cellulose claim 1 , comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'preparing the composition according to , incubating the composition, ...

FERMENTATIVE PRODUCTION OF CARBOHYDRATES BY MICROBIAL CELLS UTILIZING A MIXED FEEDSTOCK

Disclosed are genetically engineered microbial cells for the production of a carbohydrate of interest, wherein the microbial cells possess an increased intracellular availability of at least one sugar phosphate, and produce the carbohydrate of interest when being cultivated in a culture medium containing a mixed monosaccharide feedstock as main carbon and energy source. Also disclosed is a fermentative method of producing a carbohydrate of interest by cultivating said genetically engineered microbial cell in the presence of a mixed monosaccharide feedstock as main carbon and energy source. 1. A genetically engineered microbial cell for production of a carbohydrate of interest , wherein the microbial cell possesses an increased intracellular availability of at least one sugar phosphate as compared to a wild-type cell , and produces the carbohydrate of interest when being cultivated in a culture medium comprising a mixed monosaccharide feedstock as main carbon and energy source , wherein said mixed monosaccharide feedstock comprises glucose and at least one additional monosaccharide selected from the group consisting of fructose and galactose.2. The genetically engineered microbial cell according to claim 1 , wherein the genetically engineered microbial cell further comprises an UDP-galactose biosynthesis pathway for intracellular formation of UDP-galactose.3. The genetically engineered microbial cell according to claim 1 , wherein the genetically engineered microbial cell further comprises a GDP-fucose biosynthesis pathway for intracellular formation of GDP-L-fucose.4. The genetically engineered microbial cell according to claim 1 , wherein the genetically engineered microbial cell further comprises an UDP-N-acetylglucosamine biosynthesis pathway for intracellular formation of UDP-N-acetylglucosamine.5. The genetically engineered microbial cell according to claim 1 , wherein the genetically engineered microbial cell further comprises an CMP-N-acetylneuraminic acid/ ...

FERMENTATIVE PROCESS FOR THE MANUFACTURE OF MALTOSYL-ISOMALTOOLIGOSACCHARIDES (MIMO)

Described herein are efficient, low cost methods for making prebiotics that contain maltosyl-isomaltooligosaccharides (MIMOs), as well as compositions made by such methods. 1. A method comprising:(a) contacting dextransucrase-producing microorganism cells with an aqueous culture medium comprising a ratio of sucrose to maltose ranging from 2.0 to about 4.5 to form a fermentation mixture, where at least one source of the sucrose and the maltose in the fermentation mixture is an impure source of sucrose or maltose; and(b) fermenting the fermentation mixture for a time and under conditions to generate a composition comprising maltosyl-isomaltooligosaccharides (MIMOs) with a mass average molecular weight distribution of about 650 to 2000 daltons.2. The method of claim 1 , where the sucrose is raw sugar claim 1 , molasses claim 1 , sugar cane syrup claim 1 , sweet sorghum claim 1 , energy cane syrup claim 1 , beet sugar claim 1 , beet molasses claim 1 , sugar beet greens claim 1 , maple syrup claim 1 , algae claim 1 , or any combinations thereof.3. The method of claim 1 , where the maltose is claim 1 , or is derived from claim 1 , maltose syrup claim 1 , high maltose syrup claim 1 , malt claim 1 , saccharified starch claim 1 , corn/maize starch claim 1 , potato starch claim 1 , tapioca starch claim 1 , wheat starch claim 1 , oat starch claim 1 , millet starch claim 1 , sorghum starch claim 1 , rice starch claim 1 , arrowroot starch claim 1 , taro starch claim 1 , kudzu starch claim 1 , yam starch claim 1 , or any combination thereof.4. The method of claim 1 , where the impure source of sucrose or maltose does not contain toxic compounds claim 1 , heavy metals claim 1 , or toxic materials detectable by HPAEC-PAD claim 1 , HPLC claim 1 , ICP-MS claim 1 , or HPLC-RID.5Leuconostoc mesenteroides, Leuconostoc citreum, Leuconostoc gasicomitatum, Leuconostoc kimchi, Leuconostoc amelibiosum, Weissella confusa, Weissella cibaria, LactococcusStreptococcus mutans, ...

Process for producing a particulate composition comprising anhydrous crystalline 2-o-alpha-d-glucosyl-l-ascorbic acid

The invention provides a process for enabling the production of a particulate composition containing anhydrous crystalline ascorbic acid 2-glucoside that does not significantly cake even when the production yield of ascorbic acid 2-glucoside does not reach 35% by weight. The process for producing a particulate composition containing anhydrous crystalline ascorbic acid 2-glucoside, which comprises allowing a CGTase to act on a solution containing either liquefied starch or dextrin and L-ascorbic acid and then allowing a glucoamylase to act on the resulting solution to obtain a solution with an ascorbic acid 2-glucoside production yield of at least 27%, purifying the obtained solution to increase the ascorbic acid 2-glucoside content to a level of over 86% by weight, precipitating anhydrous crystalline ascorbic acid 2-glucoside by a controlled cooling method or pseudo-controlled cooling method, collecting the precipitated anhydrous crystalline ascorbic acid 2-glucoside, and ageing and drying the collected anhydrous crystalline ascorbic acid 2-glucoside.

PREPARATION OF POLY ALPHA-1,3-GLUCAN ETHERS

Poly alpha-1,3-glucan ether compounds are disclosed herein with a degree of substitution of about 0.05 to about 3.0. Also disclosed are methods of producing poly alpha-1,3-glucan ether compounds. 2. The composition of claim 1 , wherein said organic group is a hydroxy alkyl group claim 1 , alkyl group claim 1 , or carboxy alkyl group claim 1 , and wherein the compound contains one type of said organic group claim 1 , or two or more types of said organic group.3. The composition of claim 2 , wherein said organic group is a hydroxypropyl claim 2 , dihydroxypropyl claim 2 , hydroxyethyl claim 2 , methyl claim 2 , ethyl claim 2 , or carboxymethyl group.4. The composition of claim 2 , wherein the compound contains one type of said organic group.5. The composition of claim 2 , wherein the compound contains two or more types of said organic group.6. The composition of claim 1 , wherein the degree of substitution is about 0.2 to about 2.0.8. The method of claim 7 , wherein said alkaline conditions comprise an alkali hydroxide solution.9. The method of claim 7 , wherein the reaction comprises an organic solvent.10. The method of claim 9 , wherein the organic solvent is isopropanol.11. The method of claim 7 , wherein step (a) further comprises:(i) heating the reaction; and/or(ii) neutralizing the pH of the reaction.12. The method of claim 7 , wherein said organic group is a hydroxy alkyl group claim 7 , alkyl group claim 7 , or carboxy alkyl group claim 7 , and wherein the compound contains one type of said organic group claim 7 , or two or more types of said organic group.13. The method of claim 7 , wherein the poly alpha-1 claim 7 ,3-glucan is in a form of a slurry.14. The method of claim 13 , wherein the slurry comprises poly alpha-1 claim 13 ,3-glucan claim 13 , sucrose claim 13 , glucose claim 13 , fructose and a glucosyltransferase enzyme.15. The method of claim 7 , wherein the poly alpha-1 claim 7 ,3-glucan is in a form of a wet cake. This application claims the benefit ...

COMPOSITIONS AND METHODS FOR MAKING ALPHA-(1,2)-BRANCHED ALPHA-(1,6) OLIGODEXTRANS

Compositions for improving the health of a subject comprise alpha-(1,2)-branched alpha-(1,6) oligodextrans, preferably with an average molecular weight between about 10 kDa and 70 kDa, between about 10% and 50% alpha-(1,2)-osidic side chains, and having at least partial indigestibility in the subject. Methods for improving the health of a subject comprise administering the composition to a subject in an amount effective to improve gut health, or to prevent or treat a gastrointestinal disorder, a cholesterol-related disorder, diabetes, or obesity. Methods for making oligodextrans having controlled size and controlled degree of branching comprise providing alpha-(1,6) oligodextrans having an average molecular weight between 0.5 and 100 kDa and introducing at least 10% alpha-(1,2)-osidic side chains onto the alpha-(1,6) oligodextrans. 1. A composition for improving the health of a subject comprising an alpha-(1 ,2)-branched alpha-(1 ,6) oligodextran.2. The composition of claim 1 , wherein said oligodextran is a prebiotic compound.3. The composition of claim 1 , wherein said oligodextran comprises at least 10% alpha-(1 claim 1 ,2)-osidic side chains.4. The composition of claim 1 , wherein said oligodextran comprises a substantially linear backbone comprising at least two alpha-D-glucopyranosyl units linked by alpha-(1 claim 1 ,6)-linkages.5. The composition of claim 4 , wherein said backbone comprises at least 90% alpha-(1 claim 4 ,6)-D-glucopyranosidic linkages.6. The composition of claim 4 , wherein said backbone has an average molecular weight between about 0.5 kDa and 100 kDa.7. The composition of claim 4 , wherein said backbone has an average molecular weight between about 10 kDa and 70 kDa and said oligodextran comprises between about 10% and 50% alpha-(1 claim 4 ,2)-osidic side chains.8. The composition of claim 1 , wherein said oligodextran comprises less than 10% alpha-(1 claim 1 ,4)-linkages.9. A method for improving the health of a subject comprising ...

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. 1. An enzymatic reaction comprising water , glucose-1-phosphate , cellodextrin , and 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 ,wherein the cellodextrin phosphorylase enzyme synthesizes insoluble cellulose.2. The enzymatic reaction of claim 1 , wherein said cellulose has a weight-average degree of polymerization (DP) of 10 to 1000.3. The enzymatic reaction of claim 1 , wherein said cellulose has a weight-average degree of polymerization (DP) of 10 to 30.4. The enzymatic reaction of claim 1 , wherein the cellodextrin comprises cellobiose.5. A method for producing insoluble cellulose claim 1 , said method comprising:a) contacting at least water, glucose-1-phosphate, cellodextrin, and 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, wherein insoluble cellulose is produced; andb) optionally, isolating the insoluble cellulose produced in step (a).6. The method of claim 5 , wherein the cellulose produced in step (a) has a weight-average degree of polymerization (DP) of about 10 to about 1000.7. The method of claim 5 , wherein the cellulose produced in step (a) has a weight-average degree of polymerization (DP) of about 10 to about 30.8. The method of claim 5 , wherein the cellulose produced in step (a) has a cellulose II crystal structure.9. The method of claim 5 , wherein the cellodextrin comprises cellobiose.10. The method of claim 5 , wherein said glucose-1-phosphate is provided in step (a) by providing a second reaction claim 5 , wherein the products of the second reaction ...