Настройки

Укажите год
-

Небесная энциклопедия

Космические корабли и станции, автоматические КА и методы их проектирования, бортовые комплексы управления, системы и средства жизнеобеспечения, особенности технологии производства ракетно-космических систем

Подробнее
-

Мониторинг СМИ

Мониторинг СМИ и социальных сетей. Сканирование интернета, новостных сайтов, специализированных контентных площадок на базе мессенджеров. Гибкие настройки фильтров и первоначальных источников.

Подробнее

Форма поиска

Поддерживает ввод нескольких поисковых фраз (по одной на строку). При поиске обеспечивает поддержку морфологии русского и английского языка
Ведите корректный номера.
Ведите корректный номера.
Ведите корректный номера.
Ведите корректный номера.
Укажите год
Укажите год
Применить Всего найдено 7249. Отображено 100.
12-01-2012 дата публикации

Biotechnological Production of Chondroitin

Номер: US20120010399A1
Автор: Antonio Trilli, Francesca Bagatin, Immacolata Busiello, Simona Daly
Принадлежит: GNOSIS SPA

Chondroitin is produced by culturing a recombinant microorganism which is obtained by inactivation of a gene encoding an enzyme responsible for addition of fructose residues to the linear chondroitin polysaccharide in a microorganism producing a fructosylated derivative of chondroitin.

Подробнее
Номер записи: 1
03-05-2012 дата публикации

Compositions and methods for modifying cell surface glycans

Номер: US20120107281A1
Автор: Robert Sackstein
Принадлежит: Individual

Methods and compositions for modifying glycans (e.g., glycans expressed on the surface of live cells or cell particles) are provided herein.

Подробнее
Номер записи: 2
24-05-2012 дата публикации

Mycobacterium mutants for vaccines with improved protective efficacy

Номер: US20120128720A1
Автор: Anjana Batni, Christiane Huygen, Nele Festjens, Nico Callewaert
Принадлежит: Individual

Tuberculosis (TB) is a major health problem and currently, the only licensed TB vaccine is Mycobacterium bovis Bacille Calmette-Guerin ( M. bovis BCG). In the present invention, mutation of mycobacterial components reportedly involved in phagosome maturation inhibition was evaluated for vaccine purposes, as such mutations should result in better vaccine antigen processing and presentation. Thus, BCG mutants in genes coding for ManLAM capping α-1,2-mannosyltransferases and the PI3P phosphatase SapM were evaluated as TB vaccines in a stringent mouse model. Vaccination with both ManLAM capping mutants and the SapM mutant resulted in significantly longer survival as compared to non-vaccinated mice, whereas vaccination with the parental BCG did not. Moreover, mice vaccinated with the SapM mutant survived significantly longer than mice vaccinated with the parental BCG. The mutant BCG strains showed unaltered phagocytosis, replication, lysosome colocalization and oxidant activity in macrophages and similarly induced autophagy in the latter. Additionally, replication and granuloma formation in mice was unaffected, indicating BCG-equivalent safety of these vaccines.

Подробнее
Номер записи: 3
19-07-2012 дата публикации

Glycosyltransferase promoter

Номер: US20120185975A1
Автор: Cheng-Li Fang, Cheng-Yu Lee, Jui-Hung Yen, Pei-Hsuan Wei, Shu-Jiau Chiou
Принадлежит: Industrial Technology Research Institute ITRI

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

Подробнее
Номер записи: 4
20-09-2012 дата публикации

Mammalian-type glycosylation in plants

Номер: US20120237972A1
Автор: Hendrik Jan Bosch, Hendrikus Antonius Cornelis Bakker
Принадлежит: Stichting Dienst Landbouwkundig Onderzoek DLO

The invention relates to the field of glycoprotein processing in transgenic plants used as cost efficient and contamination safe factories for the production of recombinant biopharmaceutical proteins or pharmaceutical compositions comprising these. The invention provides a plant comprising a functional mammalian enzyme providing N-glycan biosynthesis that is normally not present in plants, said plant additionally comprising at least a second mammalian protein or functional fragment thereof that is normally not present in plants.

Подробнее
Номер записи: 5
08-11-2012 дата публикации

Production of Multi-Antennary N-Glycan Structures in Plants

Номер: US20120283420A1
Автор: Bieke Nagels, Koen Weterings
Принадлежит: Individual

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

Подробнее
Номер записи: 6
31-01-2013 дата публикации

Thermostable sucrose phosphorylase

Номер: US20130029384A1
Автор: An Cerdobbel, Tom Desmet, Wim Soetaert
Принадлежит: Universiteit Gent

The present invention relates to a sucrose phosphorylase from Bifidobacterium adolescentis which is useful as a biocatalyst in carbohydrate conversions at high temperatures. Indeed, the biocatalysts of the present invention are enzymatically active for a time period of at least 16 h and up to 1 to 2 week(s) at a temperature of at least 60° C. The biocatalysts of the present invention are: a) immobilized on an enzyme carrier, or b) are part of a cross-linked enzyme aggregate (CLEA), and/or c) are mutated, and/or d) are enzymatically active in the continuous presence of their substrate.

Подробнее
Номер записи: 7
07-02-2013 дата публикации

Cells, nucleic acids, enzymes and use thereof, and methods for the production of sophorolipids

Номер: US20130035403A1
Автор: Anja Thiessenhusen, Mirja Wessel, Steffen Schaffer
Принадлежит: EVONIK DEGUSSA GmbH

The invention relates to cells, nucleic acids, and enzymes, the use thereof for producing sophorolipids, and methods for producing sophorolipids.

Подробнее
Номер записи: 8
28-02-2013 дата публикации

Preparation method of virus expressing alpha-galactose epitope and vaccine

Номер: US20130052219A1
Автор: Haruko Ogawa, Kunitoshi Imai, Takahiro Tagami
Принадлежит: NATIONAL AGRICULTURE AND FOOD RESEARCH ORGANIZATION, Obihiro University of Agriculture and Veterinary Medicine NUC

A method for producing an α-Gal-expressing virus having enhanced immune response to viruses, without requiring the use of any enzyme; an influenza virus vaccine having a high effect (antigenicity), which is produced using an α-Gal-expressing virus produced by the method; and others. Specifically disclosed are: a method for producing an α-galactose epitope (Galα1-3Galβ1-4GlcNAc-R: α-Gal hereinafter)-expressing virus, which comprises the steps of: (1) introducing an α1,3- galactosyltransferase gene in an expressible condition into a cell line that does not express α-Gal to obtain a cell line capable of expressing α-Gal; (2) inoculating a virus into the cell line capable of expressing α-Gal to obtain a cell line infected with a virus; and (3) culturing the cell line infected with the virus to obtain a virus expressing α-Gal from the culture medium..

Подробнее
Номер записи: 9
28-02-2013 дата публикации

Production of galactosylated n-glycans in plants

Номер: US20130052683A1
Автор: Koen Weterings, Sylvie Van Herrewege
Принадлежит: Icon Genetics AG

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

Подробнее
Номер записи: 10
28-02-2013 дата публикации

Yeast strains producing mammalian-like complex n-glycans

Номер: US20130053550A1
Автор: Steven Christian Jozef Geysens, Wouter Vervecken
Принадлежит: Oxyrane UK Ltd

Described herein are methods and genetically engineered fungal cells useful for producing target molecules containing mammalian-like complex N-glycans or containing intermediates in a mammalian glycosylation pathway.

Подробнее
Номер записи: 11
23-05-2013 дата публикации

Cells and methods for producing rhamnolipids

Номер: US20130130319A1
Автор: Anja Thiessenhusen, Mirja Wessel, Nadine Stein, Steffen Schaffer
Принадлежит: EVONIK GOLDSCHMIDT GMBH

The invention relates to cells and nucleic acids and also use thereof for producing rhamnolipids, and also methods for producing rhamnolipids.

Подробнее
Номер записи: 12
30-05-2013 дата публикации

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

Номер: US20130137150A1
Автор: Scott E. Baker, Ziyu Dai
Принадлежит: Battelle Memorial Institute Inc

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

Подробнее
Номер записи: 13
15-08-2013 дата публикации

FUCOSYL TRANSFERASE GENE

Номер: US20130212740A1
Автор: ALTMANN Friedrich, Glossl Josef, Leiter Haralt, Mucha Jan, Staudacher Erika
Принадлежит:

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

Подробнее
Номер записи: 14
22-08-2013 дата публикации

NOVEL FUCOSYLTRANSFERASES AND THEIR APPLICATIONS

Номер: US20130217068A1
Автор: Elling Lothar, Engels Leonie, Hüfner Eric, Jennewein Stefan, Parkot Julia
Принадлежит: JENNEWEIN BIOTECHNOLOGIE GMBH

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 ...

Подробнее
Номер записи: 15
19-09-2013 дата публикации

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

Номер: US20130243690A1
Автор: Boopathy Ramakrishnan, Elizabeth Boeggeman, Marta Pasek, Pradman K. Qasba
Принадлежит: US Department of Health and Human Services

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

Подробнее
Номер записи: 16
03-10-2013 дата публикации

Modified polynucleotides for the production of secreted proteins

Номер: US20130259923A1
Автор: Antonin De Fougerolles, Atanu Roy, Jason P. SCHRUM, Jeff Lynn Ellsworth, Justin Guild, Kenechi Ejebe, Kristy M. WOOD, Matthias John, Paul Hatala, Sayda M. ELBASHIR, Stephane Bancel, Susan Whoriskey, Tirtha Chakraborty
Принадлежит: Moderna Therapeutics Inc

The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

Подробнее
Номер записи: 17
07-11-2013 дата публикации

Synthetic genes and genetic constructs

Номер: US20130298264A1
Автор: Michael Wayne Graham, Robert Norman Rice
Принадлежит: Commonwealth Scientific and Industrial Research Organization CSIRO

The present invention relates generally to synthetic genes for modifying endogenous gene expression in a cell, tissue or organ of a transgenic organism, in particular a transgenic animal or plant. More particularly, the present invention provides novel synthetic genes and genetic constructs which are capable of repressing delaying or otherwise reducing the expression of an endogenous gene or a target gene in an organism when introduced thereto.

Подробнее
Номер записи: 18
14-11-2013 дата публикации

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

Номер: US20130302857A1
Автор: Chen Chao, DESMET Tom, Soetaert Wim, Van Der Borght Jef
Принадлежит: Universiteit Gent

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

Подробнее
Номер записи: 19
21-11-2013 дата публикации

METHODS FOR ALTERING THE REACTIVITY OF PLANT CELL WALLS

Номер: US20130312141A1
Автор: De Block Marc, Essigmann Bernd, KOCH Rainhard, Meulewaeter Frank
Принадлежит: Bayer BioScience N.V.

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

Подробнее
Номер записи: 20
28-11-2013 дата публикации

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

Номер: US20130316406A1
Автор: Apel William A., Henriksen Emily D., Lacey Jeffrey A., Reed David W., Thompson David N., Thompson Vicki S.
Принадлежит: BATTELLE ENERGY ALLIANCE, LLC

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

Подробнее
Номер записи: 21
12-12-2013 дата публикации

COMPOSITIONS RELATING TO A MUTANT CLOSTRIDIUM DIFFICILE TOXIN AND METHODS THEREOF

Номер: US20130330371A1
Автор: Anderson Annaliesa Sybil, Donald Robert G. K., Flint Michael James, Jansen Kathrin Ute, Kalyan Narender Kumar, Moran Justin Keith, Ruppen Mark E., Sidhu Maninder K.
Принадлежит: WYETH LLC

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

Подробнее
Номер записи: 22
12-12-2013 дата публикации

Fusion Enzymes

Номер: US20130330780A1
Автор: Hiltunen Jukka, Huuskonen Anne, Kanerva Anne, Natunen Jari, Saloheimo Markku, Viskari Heli
Принадлежит:

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 ...

Подробнее
Номер записи: 23
19-12-2013 дата публикации

Method of producing succinic acid and other chemicals using sucrose-containing feedstock

Номер: US20130337519A1
Автор: R. Rogers Yocum, Sudhandshu Dole, Theron Hermann, Xiaohui Yu
Принадлежит: Myriant Corp

This invention relates to the production of chemicals by fermentation with a microorganism in which the fermentation medium contains the sugar sucrose. As a specific example, succinic acid is produced from a sucrose-containing renewable feedstock through fermentation using a biocatalyst. Examples of such a biocatalyst include microorganisms that have been enhanced in their ability to utilize sucrose as a carbon and energy source. The biocatalysts of the present invention are derived from the genetic manipulation of parental strains that were originally constructed with the goal to produce one or more chemicals (for example succinic acid and/or a salt of succinic acid) at a commercial scale using feedstocks other than sucrose. The genetic manipulations of the present invention involve the introduction of exogenous genes involved in the transport and metabolism of sucrose into the parental strains. The genes involved in the transport and metabolism of sucrose can also be introduced into a microorganism prior to developing the organism to produce a particular chemical. The genes involved in the transport and metabolism of sucrose can also be used to augment or improve the sucrose transport and metabolism by strains already known to have some ability for sucrose utilization in biological fermentation.

Подробнее
Номер записи: 24
26-12-2013 дата публикации

GNTIII EXPRESSION IN PLANTS

Номер: US20130347146A1
Автор: Bakker Hendrikus Antonius Cornelis, Bosch Hendrik Jan, Florack Dionisius Elisabeth Antonius
Принадлежит: Stichting Dienst Landbouwkundig Onderzoek

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

Подробнее
Номер записи: 25
09-01-2014 дата публикации

MALTOTRIOSYL TRANSFERASE, PROCESS FOR PRODUCTION THEREOF, AND USE THEREOF

Номер: US20140011257A1
Автор: Nagaya Miho, Okada Masamichi, Yamaguchi Shotaro
Принадлежит: AMANO ENZYME INC.

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

Подробнее
Номер записи: 26
16-01-2014 дата публикации

Method for utilizing monoterpene glycosyltransferase

Номер: US20140020137A1
Автор: Eiichiro Ono, Nobuo Tsuruoka
Принадлежит: Suntory Holdings Ltd

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.

Подробнее
Номер записи: 27
23-01-2014 дата публикации

NOVEL FUCOSYLTRANSFERASES AND THEIR APPLICATIONS

Номер: US20140024820A1
Автор: Elling Lothar, Engels Leonie, Huefner Eric, Jennewein Stefan, Parkot Julia
Принадлежит: JENNEWEIN BIOTECHNOLOGIE GMBH

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- ...

Подробнее
Номер записи: 28
20-02-2014 дата публикации

HIGH LEVEL EXPRESSION OF RECOMBINANT TOXIN PROTEINS

Номер: US20140051093A1
Автор: Chew Lawrence, Retallack Diane M.
Принадлежит: PFENEX INC.

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

Подробнее
Номер записи: 29
13-03-2014 дата публикации

Biotechnological Production of Chondroitin

Номер: US20140073772A1
Автор: Antonio Trilli, Francesca Bagatin, Immacolata Busiello, Simona Daly
Принадлежит: GNOSIS SPA

Chondroitin is produced by culturing a recombinant microorganism which is obtained by inactivation of a gene encoding an enzyme responsible for addition of fructose residues to the linear chondroitin polysaccharide in a microorganism producing a fructosylated derivative of chondroitin.

Подробнее
Номер записи: 30
27-03-2014 дата публикации

Glucosyltransferase enzymes for production of glucan polymers

Номер: US20140087431A1
Автор: Hongxian He, Mark S. Payne, Thomas Scholz, Yefim Brun
Принадлежит: EI Du Pont de Nemours and Co

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.

Подробнее
Номер записи: 31
06-01-2022 дата публикации

Compositions and Production of Recombinant AAV Viral Vectors Capable of Glycoengineering In Vivo

Номер: US20220002387A1
Автор: Desrosiers Ronald, Termini James M.
Принадлежит:

The disclosure provides an expression vector (e.g., AAV vector) comprising a nucleic acid sequence encoding (1) the heavy and/or light chain of an antibody and (2) one or more shRNA sequences targeting fucosyltransferase-8 (FUT8). 1. An expression vector comprising a nucleic acid sequence encoding (1) the heavy and/or light chain of an antibody and (2) one or more shRNA sequences targeting fucosyltransferase-8 (FUT8).2. The expression vector of claim 1 , wherein the expression vector encodes both the heavy chain and the light chain of an antibody.3. The expression vector of claim 1 , wherein the expression vector is an adeno-associated viral (AAV) vector.4. The expression vector of claim 1 , wherein the heavy chain comprises one or more mutations in the Fc region which enhances antibody-dependent cell cytotoxicity.5. The expression vector of claim 4 , wherein mutation is an LS mutation (M428L/N434S) claim 4 , a LALA mutation (L234A claim 4 , L235A) claim 4 , a S239 (DFL) mutation (S239D/1332F/A330L) a C6A-74 mutation (V259I/N315D/N434Y) claim 4 , a HN mutation (H433K/N434F) claim 4 , K392D/K409D/A330M/K334V claim 4 , E356K/D399K/L234Y/Y296W claim 4 , K392D/K409D/S239D/A330M/K334V claim 4 , E356K/D399K/L234Y/K290Y/Y296W claim 4 , K392D/K409D/A330M/K334V claim 4 , E356K/D399K/L234Y/K290Y/Y296W claim 4 , K392D/K409D/A330F/K334V claim 4 , E356K/D399K/L234Y/K290Y/Y296W claim 4 , K392D/K409D/A330M/K334V claim 4 , or E356K/D399K/K290Y/Y296W.6. A composition comprising (a) an expression vector comprising a nucleic acid sequence encoding a heavy chain of an antibody and (b) an expression vector comprising a nucleic acid sequence encoding a light chain of an antibody claim 4 , wherein (a) claim 4 , (b) claim 4 , or (a) and (b) further comprises one or more shRNA sequences targeting fucosyltransferase-8 (FUT8).7. The composition of claim 6 , wherein the expression vector is an adeno-associated viral (AAV) vector.8. The composition of claim 6 , wherein the heavy chain comprises ...

Подробнее
Номер записи: 32
06-01-2022 дата публикации

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

Номер: US20220002773A1
Автор: Beauprez Joeri, Lannoo Nausicaä, Vandewalle Kristof, Vercauteren Annelies
Принадлежит:

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 ...

Подробнее
Номер записи: 33
07-01-2016 дата публикации

POLYPEPTIDE

Номер: US20160002659A1
Автор: BULONE Vincent, CAMPILHO Ana, CARLSBECKER Annelie, DETTMER Jan, FURUTA Kaori, HELARIUTTA Yrjö, LEHESRANTA Satu, LICHTENBERGER Raffael, MÄHÖNEN Ari Pekka, MIYASHIMA Shunsuke, VATÈN Anne, YADAV Shri Ram
Принадлежит:

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

Подробнее
Номер записи: 34
07-01-2016 дата публикации

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

Номер: US20160002693A1
Автор: CAIMI PERRY G., Hennessey Susan Marie, PAYNE MARK S.
Принадлежит:

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 ...

Подробнее
Номер записи: 35
05-01-2017 дата публикации

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

Номер: US20170002335A1
Автор: Bott Richard R, Brun Yefim, PAYNE MARK S
Принадлежит:

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 ...

Подробнее
Номер записи: 36
05-01-2017 дата публикации

Modified glucosyltransferases for producing branched alpha-glucan polymers

Номер: US20170002336A1
Автор: Mark S. Payne, Richard R. Bott, Yefim Brun
Принадлежит: DANISCO US INC, EI Du Pont de Nemours and Co

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.

Подробнее
Номер записи: 37
05-01-2017 дата публикации

Fusion Enzymes

Номер: US20170002337A1
Автор: Hiltunen Jukka, Huuskonen Anne, Kanerva Anne, Natunen Jari, Saloheimo Markku, Viskari Heli
Принадлежит: GLYKOS FINLAND OY

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

Подробнее
Номер записи: 38
05-01-2017 дата публикации

Pichia pastoris surface display system

Номер: US20170002346A1
Автор: Kurt R. Gehlsen, Thomas G. Chappell
Принадлежит: Research Corp Technologies Inc

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

Подробнее
Номер записи: 39
07-01-2021 дата публикации

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

Номер: US20210002382A1
Автор: BRÜNKER Peter, FERRARA KOLLER Claudia, Mössner Ekkehard, PÜNTENER Ursula, SUTER Tobias, Umana Pablo
Принадлежит: ROCHE GLYCART AG

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

Подробнее
Номер записи: 40
07-01-2021 дата публикации

Crispr/cas9 vector combination and application thereof in gene knockout

Номер: US20210002652A1
Автор: Haiyuan Yang, Yifan Dai, YING Wang
Принадлежит: Nanjing Genefriend Biotech Inc

Provided is an SgRNA combination, comprising an SgRNA specifically targeting the GGTA1 gene, an SgRNA specifically targeting the CMAH gene and an SgRNA specifically targeting the β4GalNT2 gene. Also provided is a CRISPR/Cas9 vector combination, comprising a GGTA1-CRISPR/Cas9 vector, a CMAH-CRISPR/Cas9 vector and a β4GalNT2-CRISPR/Cas9 vector. Also provided is an applicaton of the CRISPR/Cas9 vector combination in knocking out the GGTA1 gene, the CMAH gene and the β4GalNT2 gene. The knockout rates of the three genes with the specifically targeted SgRNA sequences are respectively 56%, 63%, and 41%. A three genes knockoutpig can be obtained, wherein the three genes related to immune rejectionare knocked out, and heart valves of said pig can be acquired.

Подробнее
Номер записи: 41
03-01-2019 дата публикации

HETERO-TRANSGLYCOSYLASE AND USES THEREOF

Номер: US20190002849A1
Автор: Frankova Lenka, Fry Stephen C., Holland Claire, Hudson Andrew, Meulewaeter Frank, Mohler Kyle E., Simmons Tom J., Van Den Brande Ilse
Принадлежит:

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 ...

Подробнее
Номер записи: 42
01-01-2015 дата публикации

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Номер: US20150004649A1
Автор: Brun Yefim, HE HONGXIAN, PAYNE MARK S., Scholz Thomas
Принадлежит:

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- ...

Подробнее
Номер записи: 43
01-01-2015 дата публикации

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Номер: US20150004650A1
Автор: Brun Yefim, HE HONGXIAN, PAYNE MARK S., Scholz Thomas
Принадлежит:

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- ...

Подробнее
Номер записи: 44
01-01-2015 дата публикации

Glucosyltransferase enzymes for production of glucan polymers

Номер: US20150004651A1
Автор: Hongxian He, Mark S Payne, Thomas Scholz, Yefim Brun
Принадлежит: EI Du Pont de Nemours and Co

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.

Подробнее
Номер записи: 45
01-01-2015 дата публикации

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Номер: US20150004652A1
Автор: Brun Yefim, HE HONGXIAN, PAYNE MARK S., Scholz Thomas
Принадлежит:

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- ...

Подробнее
Номер записи: 46
01-01-2015 дата публикации

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Номер: US20150004653A1
Автор: Brun Yefim, HE HONGXIAN, PAYNE MARK S., Scholz Thomas
Принадлежит:

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- ...

Подробнее
Номер записи: 47
13-01-2022 дата публикации

GLYCOGEN-NULL METHANOTROPHS AND USES THEREOF

Номер: US20220010268A1
Автор: Hsi Megan, Luning Eric G., Rueda Paloma, Saville Renee M., Silverman Joshua A., Stegantseva Yelena, Tang Vincent
Принадлежит:

The present disclosure provides methanotrophic bacteria that are modified to produce less glycogen, and methods of using the modified methanotrophic bacteria to produce a desired product, such as protein(s) or metabolite(s). 1Methylococcus capsulatusMethylococcus capsulatus. A modified , comprising a chromosomal knock-out of an ADP-glucose pyrophosphorylase gene , a glgA2 isoform of a glycogen synthase gene , or both , wherein the modified cultured under conditions comprising a non-limiting amount of a Csubstrate produces:{'i': 'Methylococcus capsulatus', 'at least about 30% less glycogen as compared to the parent cultured under the same conditions; and/or'}{'i': 'Methylococcus capsulatus', 'at least about 5% more crude protein as compared to the parent cultured under the same conditions.'}2Methylococcus capsulatusMethylococcus capsulatusMethylococcus capsulatus. The modified of claim 1 , wherein the is Bath.3Methylococcus capsulatus. The modified of or claim 1 , wherein the culture conditions further comprise the presence of a limiting amount of a nutrient or metabolite required for growth.4Methylococcus capsulatus. The modified of claim 3 , wherein the limiting amount of the nutrient required for growth comprises a limiting amount of nitrogen claim 3 , sulfur claim 3 , phosphorous claim 3 , and/or oxygen.5Methylococcus capsulatus. The modified of claim 4 , wherein the limiting amount of the nitrogen comprises a limiting amount of nitrate claim 4 , ammonium claim 4 , and/or nitrogen gas.6Methylococcus capsulatusMethylococcus capsulatus. The modified of any of - claim 4 , wherein the production of at least about 5% more crude protein as compared to the parent further comprises a culture condition comprising from about 20% to about 80% nitrogen fixation.7Methylococcus capsulatusMethylococcus capsulatusMethylococcus capsulatus.. The modified of any of - claim 4 , wherein the modified has a lower ratio of utilized oxygen to utilized methane as compared to the ...

Подробнее
Номер записи: 48
01-01-2015 дата публикации

Heat stable mutants of starch biosynthesis enzymes

Номер: US20150007363A1
Автор: L. Curtis Hannah, Thomas W. Greene
Принадлежит: University of Florida Research Foundation Inc

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

Подробнее
Номер записи: 49
12-01-2017 дата публикации

CEREAL SEED STARCH SYNTHASE II ALLELES AND THEIR USES

Номер: US20170006815A1
Автор: GIROUX Michael J.
Принадлежит:

The present invention provides compositions and methods of altering/improving wheat phenotypes. Furthermore, methods of breeding wheat and/or other closely related species to produce plants having altered or improved phenotypes are provided. 2. The high amylose grain of wherein the proportion of starch amylose of said grain is at least 25% higher compared to the starch amylose of a control grain from an appropriate wild type wheat check variety grown under similar field conditions.3. The high amylose grain of wherein said grain has at least a 10% higher seed weight than the grain from an appropriate null wheat check variety grown under similar field conditions.4. The high amylose grain of claim 1 , wherein the wheat plant is a hexaploid wheat comprising a first claim 1 , second claim 1 , and third genome.5. The high amylose grain of claim 4 , wherein the hexaploid wheat comprises homozygous SSII null alleles in the first and second genomes claim 4 , and the SSII leaky allele in the third genome.6. The high amylose grain of claim 5 , wherein the SSII leaky allele is homozygous in the third genome.7. The high amylose grain of claim 5 , wherein the SSII leaky allele comprises a missense mutation encoding for a protein with an SSII-D-E656K and/or SSII-D-A421V amino acid substitution.8. The high amylose grain of claim 5 , wherein the SSII leaky allele encodes the protein of SEQ ID No. 40 or SEQ ID No. 44.9. The high amylose grain of claim 1 , wherein the wheat plant is a tetraploid wheat comprising a first and second genome.10. The high amylose grain of claim 9 , wherein the tetraploid wheat comprises homozygous SSII null alleles in the first genome claim 9 , and the SSII leaky allele in the second genome.11. The high amylose grain of claim 10 , wherein the SSII leaky allele is homozygous in the second genome.12. The high amylose grain of claim 9 , wherein the SSII leaky allele comprises a missense mutation encoding for a protein with an SSII-B-P333L and/or SSII-B-P333S ...

Подробнее
Номер записи: 50
12-01-2017 дата публикации

STRATEGY FOR SUCROSE REDUCTION AND GENERATION OF INSOLUBLE FIBER IN JUICES

Номер: US20170006902A1
Автор: Garske Adam L.
Принадлежит: DANISCO US INC.

The present teachings provide a method of making a lower calorie, higher insoluble fiber beverage comprising; treating a sucrose-containing beverage with a glucosyltransferase to convert sucrose to alpha (1-3) glucan to make the lower calorie, higher insoluble fiber beverage. Additional methods, as well as compositions, are provided. 1. A method of making a lower calorie , higher insoluble fiber beverage comprising;treating a sucrose-containing beverage with a glucosyltransferase to convert sucrose to alpha (1-3) glucan to make the lower calorie, higher insoluble fiber beverage.2. The method of claim 1 ,further comprising, removing the alpha (1-3) glucan to make a lower calorie, clarified beverage.3. The method of claim 1 , wherein the lower calorie claim 1 , higher insoluble fiber beverage contains no less than 95% claim 1 , 96% claim 1 , 97% claim 1 , 98% claim 1 , 99% claim 1 , or 99.9% fructose.4. The method according to wherein the treating comprises treating with a glucosyltransferase comprising SEQ ID NO: 1.5. The method according to wherein the treating comprises treating with an enzyme 70% claim 1 , 75% claim 1 , 80% claim 1 , 85% claim 1 , 90% claim 1 , 95% claim 1 , 98% claim 1 , 99% claim 1 , or 99.9% identical to SEQ ID NO:1.6. The method according to wherein the treating comprises treating with an immobilized glucosyltransferase.7Streptococcus salivarius.. The method according to wherein the treating comprises treating with an immobilized glucosyltransferase from8. The method according to wherein the beverage comprises a fruit juice.9. The method according to wherein the beverage comprises apple juice or orange juice.10. The method according to wherein the beverage comprises a sucrose level reduced at least 30% claim 1 , at least 40% claim 1 , at least 50% claim 1 , at least 60% claim 1 , at least 70% claim 1 , at least 80% claim 1 , at least 85% claim 1 , or at least 90% compared to a control beverage lacking the treating.11. A beverage comprising at ...

Подробнее
Номер записи: 51
20-01-2022 дата публикации

Efficient Production of Steviol Glycosides in Recombinant Hosts

Номер: US20220017932A1
Автор: Anderson Iben Nordmark, Dalgaard Mikkelsen Michael, Douchin Veronique, Hansen Jorgen, Simon Ernesto
Принадлежит:

Recombinant microorganisms are disclosed that produce steviol glycosides and have altered expression of one or more endogenous transporter or transcription factor genes, or that overexpress one or more heterologous transporters, leading to increased excretion of steviol glycosides of interest. 1. A recombinant microorganism capable of producing a steviol glycoside in a cell culture ,wherein the microorganism has a modified expression of at least one endogenous transporter gene and at least one endogenous transcription factor gene that regulates expression of the at least one endogenous transporter gene,wherein the modified expression comprises reduced expression or activity of the at least one endogenous transporter gene, having at least 80% sequence identity to the amino acid sequence set forth in SEQ ID NO:113;wherein the reduced expression is produced in the microorganism by disrupting or deleting the gene locus for the at least one endogenous transporter gene;wherein the microorganism further expresses at least one heterologous transporter gene;wherein the steviol glycoside is Rebaudioside A, Rebaudioside B, Rebaudioside D, Rebaudioside E, Rebaudioside M, or an isomer thereof.2. The recombinant microorganism of claim 1 , wherein the endogenous and/or the heterologous transporter gene encodes an ATP-Binding Cassette (ABC) transporter.3Stevia. The recombinant microorganism of claim 1 , wherein the heterologous transporter gene is a transporter gene.4. The recombinant microorganism of claim 1 , further comprising a gene encoding a polypeptide capable of beta 1 claim 1 ,2 glycosylation of the C2′ of the 13-O-glucose claim 1 , 19-O-glucose claim 1 , or both 13-O-glucose and 19-O-glucose of a steviol glycoside; wherein the gene has a copy number of 2 or more; and wherein the polypeptide comprises a polypeptide having at least 90% sequence identity to one of the amino acid sequences set forth in any one of SEQ ID NOs:51 claim 1 , 54 claim 1 , 55 claim 1 , 86 claim 1 , ...

Подробнее
Номер записи: 52
12-01-2017 дата публикации

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

Номер: US20170009266A1
Автор: Floris Louis Van Delft, Maria Antonia WIJDEVEN, Remon VAN GEEL, Ryan HEESBEEN
Принадлежит: Synaffix BV

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.

Подробнее
Номер записи: 53
08-01-2015 дата публикации

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Номер: US20150010954A1
Автор: Brun Yefim, HE HONGXIAN, PAYNE MARK S., Scholz Thomas
Принадлежит:

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- ...

Подробнее
Номер записи: 54
08-01-2015 дата публикации

Glucosyltransferase enzymes for production of glucan polymers

Номер: US20150010956A1
Автор: Hongxian He, Mark S. Payne, Thomas Scholz, Yefim Brun
Принадлежит: EI Du Pont de Nemours and Co

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.

Подробнее
Номер записи: 55
11-01-2018 дата публикации

NON-CALORIC SWEETENER

Номер: US20180009835A1
Автор: CHATURVEDULA VENKATA SAI PRAKASH, Mao Guohong, Yu Xiaodan
Принадлежит: Conagen Inc.

Disclosed is a steviol glycoside referred to as rebaudioside D2. Rebaudioside D2 has five β-D-glucosyl units connected to the aglycone steviol. Also disclosed are methods for producing rebaudioside D2, a UDP-glycosyltransferase fusion enzyme, and methods for producing rebaudioside D and rebaudioside E. 144-. (canceled)45. A method for synthesizing rebaudioside E , the method comprising:preparing a reaction mixture comprising stevioside; a substrate selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose); and a UDP-glycosyltransferase selected from the group consisting of a uridine diphospho glycosyltransferase and a UDP-glycosyltransferase fusion enzyme comprising a uridine diphospho glycosyltransferase domain coupled to a sucrose synthase domain, wherein a glucose is covalently coupled to the stevioside to produce rebaudioside E.46Oryza sativa. The method of claim 45 , wherein the UDP-glycosyltransferase is a uridine diphospho glycosyltransferase EUGT11.47. The method of further comprising adding a sucrose synthase to the reaction mixture.48ArabidopsisCoffeaStevia. The method of claim 47 , wherein the sucrose synthase is selected from the group consisting of an sucrose synthase 1; a sucrose synthase 1 and a sucrose synthase 1.49Arabidopsis thaliana. The method of claim 47 , wherein the sucrose synthase is an sucrose synthase 1.50. The method of claim 45 , wherein the UDP-glycosyltransferase is a UDP-glycosyltransferase fusion enzyme comprising a uridine diphospho glycosyltransferase domain coupled to a sucrose synthase domain.51. The method of claim 50 , wherein the UDP-glycosyltransferase fusion enzyme comprises an amino acid sequence having about 90% sequence identity to SEQ ID NO:5.52Oryza sativa. The method of claim 45 , wherein the uridine diphospho glycosyltransferase domain is an uridine diphospho glycosyltransferase EUGT11.53. The method of claim 45 , wherein the uridine diphospho ...

Подробнее
Номер записи: 56
11-01-2018 дата публикации

IMMUNOGENIC BACTERIAL VESICLES WITH OUTER MEMBRANE PROTEINS

Номер: US20180009852A1
Автор: ADU-BOBIE Jeannette, FERRARI Germano, Grandi Guido, NORAIS Nathalie, Pizza Mariagrazia
Принадлежит: GLAXOSMITHKLINE BIOLOGICALS SA

Knockout of the meningococcal mltA homolog gives bacteria that spontaneously release vesicles that are rich in immunogenic outer membrane proteins and that can elicit cross-protective antibody responses with higher bactericidal titres than OMVs prepared by normal production processes. Thus the invention provides a bacterium having a knockout mutation of its mltA gene. The invention also provides a bacterium, wherein the bacterium: (i) has a cell wall that includes peptidoglycan; and (ii) does not express a protein having the lytic transglycosylase activity MltA protein. The invention also provides compositions comprising vesicles that, during culture of bacteria of the invention, are released into the culture medium. 125-. (canceled)26Escherichia bacterium. A process for preparing bacterial vesicles , comprising the steps of: (i) culturing an in a culture medium such that the bacterium releases vesicles into said medium; and (ii) collecting the vesicles from said medium , wherein: (a) the bacterium has a cell wall that includes peptidoglycan; and (b) the bacterium does not express a protein having the lytic transglycosylase activity of MltA protein.27. The process of claim 26 , wherein the bacterium also has a knockout mutation of at least one further gene.28E. coli.. The process of claim 26 , wherein the bacterium is29E. coli.. The process of claim 28 , wherein the bacterium is a pathogenic30E. coli. The process of claim 29 , wherein the pathogenic is an extraintestinal pathogenic bacterium claim 29 , a uropathogenic bacterium claim 29 , or a meningitis/sepsis-associated bacterium.31E. coli.. The process of claim 27 , wherein the bacterium is32E. coli.. The process of claim 31 , wherein the bacterium is a pathogenic33E. coli. The process of claim 32 , wherein the pathogenic is an extraintestinal pathogenic bacterium claim 32 , a uropathogenic bacterium claim 32 , or a meningitis/sepsis-associated bacterium.34Escherichia coli. The process of claim 26 , wherein the ...

Подробнее
Номер записи: 57
14-01-2021 дата публикации

URIDINE DIPHOSPHATE-DEPENDENT GLYCOSYLTRANSFERASE ENZYME

Номер: US20210009968A1
Автор: DONALD Jason Eric, KUMARAN Ajikumar Parayil, LOVE Aaron, SANTOS Christine Nicole S., TOOMEY Christopher
Принадлежит:

In various aspects, the present invention provides uridine diphosphate-dependent glycosyltransferase (UGT) enzymes capable of catalyzing the transfer of a monosaccharide moiety from a NDP-sugar to the 3′ carbon of a sugar moiety of a substrate, such as a terpenoid glycan, thereby functioning as a “1-3 UGT.” In other aspects, the invention provides polynucleotides encoding the 1-3 UGT, and host cells comprising the same. In still other aspects, the invention provides methods for preparing glycosylated substrates, including steviol glycosides, using the enzyme and host cells of this disclosure. 1. A uridine diphosphate-dependent glycosyltransferase (UGT) enzyme comprising an amino acid sequence that is at least about 75% identical to the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 6.2. The enzyme of claim 1 , wherein the amino acid sequence is at least about 80% identical to SEQ ID NO: 5 or SEQ ID NO: 6.3. The enzyme of claim 1 , wherein the amino acid sequence is at least about 85% identical to SEQ ID NO: 5 or SEQ ID NO: 6.4. The enzyme of claim 1 , wherein the amino acid sequence is at least about 90% identical to SEQ ID NO: 5 or SEQ ID NO: 6.5. The enzyme of claim 1 , wherein the amino acid sequence is at least about 95% identical to SEQ ID NO: 5 or SEQ ID NO: 6.6. The enzyme of claim 1 , wherein the amino acid sequence is at least about 98% identical to SEQ ID NO: 5 or SEQ ID NO: 6.7. The enzyme of claim 1 , wherein the amino acid sequence is the amino acid of SEQ ID NO: 5 or SEQ ID NO: 6.8. The enzyme of claim 1 , wherein the amino acid sequence has from 1 to 20 amino acid modifications independently selected from substitutions claim 1 , deletions claim 1 , and insertions claim 1 , with respect to the amino acid sequence SEQ ID NO: 5 or SEQ ID NO: 6.9. The enzyme of claim 8 , wherein the amino acid sequence has from 1 to 10 amino acid modifications independently selected from substitutions claim 8 , deletions claim 8 , and insertions claim 8 , with respect ...

Подробнее
Номер записи: 58
14-01-2021 дата публикации

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

Номер: US20210010056A1
Автор: WANG Xiaoqiang
Принадлежит:

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

Подробнее
Номер записи: 59
10-01-2019 дата публикации

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

Номер: US20190010532A1
Автор: Gupta Shivani, Kang Sohye
Принадлежит: Amgen Inc.

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

Подробнее
Номер записи: 60
09-01-2020 дата публикации

Engineered Cells And Methods Of Use

Номер: US20200010795A1
Автор: Chen Mingkuan, Li Jie, Wu Peng, Zhou Yiran
Принадлежит:

Provided herein are engineered cells, comprising: a chemical or biological moiety covalently bound to a cell surface glycan, wherein the chemical or biological moiety is selected from the group consisting of small molecule, polynucleotide, polypeptide, and antibody. Also provided are compositions comprising these engineered cells and methods of making and using the same. 1. An engineered cell , comprising:a chemical or biological moiety covalently bound to a cell surface glycan present on the surface of the cell,wherein the chemical or biological moiety is selected from the group consisting of small molecule, polynucleotide, polypeptide, and antibody.2. The engineered cell of claim 1 , wherein the engineered cell is an immune cell.3. The engineered cell of claim 1 , wherein the engineered cell is a T-cell or a natural killer (NK) cell.4. The engineered cell of claim 3 , wherein the T cell is a CD8+ or a CD4+ T cell.5. The engineered cell of claim 1 , wherein the cell is a Dendritic Cell (DC).6. The engineered cell of claim 1 , wherein the small molecule is a small drug molecule or a pharmaceutically acceptable salt or a co-crystal thereof.7. The engineered cell of claim 1 , wherein the antibody is a single chain variable fragment (scFv) claim 1 , fragment antigen binding (Fab) fragment claim 1 , or a full length antibody.8. The engineered cell of claim 1 , wherein the antibody is an immunoglobulin G (IgG) antibody.9. The engineered cell of claim 8 , wherein the IgG is a full length IgG.10. The engineered cell of claim 1 , wherein the chemical or biological moiety attached to the engineered cell is a biological marker and/or probe.11. The engineered cell of claim 1 , wherein the chemical or biological moiety is a biotin probe claim 1 , a fluorescent probe claim 1 , a biorthogonal reaction handle claim 1 , and/or a dye labeled single strand DNA.12. The engineered cell of claim 11 , wherein the dye is FAM.13. The engineered cell of claim 11 , wherein the fluorescent ...

Подробнее
Номер записи: 61
09-01-2020 дата публикации

ENZYMATIC PRODUCTION OF HEXOSES

Номер: US20200010824A1
Автор: ROGERS Edwin O., WICHELECKI Daniel Joseph
Принадлежит: BONUMOSE LLC

Disclosed herein are methods of producing hexoses from saccharides by enzymatic processes. The methods utilize fructose 6-phosphate and at least one enzymatic step to convert it to a hexose. 1. An enzymatic process for preparing fructose from a starch derivative , the process comprising the steps of:(i) a step of converting a starch derivative to glucose 1-phosphate (G1P) using alpha glucan phosphorylase (αGP);(ii) a step of converting G1P to glucose 6-phosphate (G6P), wherein the step is catalyzed by phosphoglucomutase (PGM)(iii) a step of converting G6P to fructose 6-phosphate (F6P), wherein the step is catalyzed by phosphoglucoisomerase (PGI)(iv) a step of converting F6P to fructose catalyzed by fructose 6-phosphate phosphatase (F6PP).2. The process of claim 1 , further comprising the step of converting starch to a starch derivative wherein the starch derivative is prepared by enzymatic hydrolysis of starch or by acid hydrolysis of starch claim 1 , wherein the enzymatic hydrolysis of starch is catalyzed by isoamylase claim 1 , pullulanase claim 1 , alpha-amylase claim 1 , or a combination thereof.3. The process of claim 1 , wherein 4-glucan transferase (4GT) is added to the process.4. The process of claim 1 , wherein the process steps are conducted under at least one of the following process conditions:at a temperature ranging from about 37° C. to about 85° C.,at a pH ranging from about 5.0 to about 9.0, orfor about 0.5 hours to about 48 hours.5. The process of claim 1 , wherein the process steps are conducted under at least one of the following process conditions:without adenosine triphosphate (ATP) as a source of phosphate,without nicotinamide adenosine dinucleotide,at a phosphate concentration from about 0.1 mM to about 150 mM,where phosphate is recycled, orwhere step (iv) involves an energetically favorable chemical reaction.6. The process of claim 5 , wherein phosphate is recycled and the phosphate produced by F6PP dephosphorylation of F6P is used in the ...

Подробнее
Номер записи: 62
19-01-2017 дата публикации

COMPOSITIONS AND PROCESSES FOR IMPROVING PROPERTIES OF FILLERS

Номер: US20170015832A1
Автор: Benyamino Romil, BERLIN Alex, DeLozier Gregory, Quinlan Jason
Принадлежит: NOVOZYMES A/S

The present invention relates to processes for modifying a filler material comprising treating the filler material with a composition comprising a xyloglucan endotransglycosylase and (a) a polymeric xyloglucan and a functionalized xyloglucan oligomer comprising a chemical group; (b) a polymeric xyloglucan functionalized with a chemical group and a functionalized xyloglucan oligomer comprising a chemical group; (c) a polymeric xyloglucan functionalized with a chemical group and a xyloglucan oligomer; (d) a polymeric xyloglucan and a xyloglucan oligomer; (e) a a polymeric xyloglucan functionalized with a chemical group; (f) a polymeric xyloglucan; (g) a functionalized xyloglucan oligomer comprising a chemical group; or (h) a xyloglucan oligomer; or a composition of (a-h) without a xyloglucan endotransglycosylase, wherein the modified filler material possesses an improved property compared to the unmodified filler material. The present invention also relates to modified filler materials and modified filler materials obtained by such processes. 1. A process for modifying a filler material comprising treating a suspension of the filler 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 composition comprising ...

Подробнее
Номер записи: 63
19-01-2017 дата публикации

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

Номер: US20170016038A1
Автор: Beauprez Joeri, De Mey Marjan, Maertens Jo
Принадлежит:

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 ...

Подробнее
Номер записи: 64
15-01-2015 дата публикации

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

Номер: US20150018522A1
Автор: Boeggeman Elizabeth, Qasba Pradman K., Ramakrishnan Boopathy
Принадлежит: The United States of America, as represented by the Secretary, Department of Health & Human Servic

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 ...

Подробнее
Номер записи: 65
21-01-2021 дата публикации

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

Номер: US20210017558A1
Автор: Beauprez Joeri, De Mey Marjan, Maertens Jo
Принадлежит:

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 ...

Подробнее
Номер записи: 66
16-01-2020 дата публикации

PRODUCTION OF STEVIOL GLYCOSIDE IN RECOMBINANT HOSTS

Номер: US20200017896A1
Автор: Carlsen Simon, Chuang Lim Swee, Dalgaard Mikkelsen Michael, Douchin Veronique, Düring Louis, Olsson Kim, Simon Ernesto
Принадлежит:

The invention relates to recombinant microorganisms and methods for producing steviol glycosides, glycosylated ent-kaurenol, and glycosylated ent-kaurenoic acid. 1. An in vitro method for producing a steviol glycoside , a glycosylated ent-kaurenol compound , and/or a glycosylated ent-kaurenoic acid compound , comprising: a first polypeptide capable of catalyzing beta 1,2 glycosylation of the C2′ of the 13-O-glucose, 19-O-glucose, or both 13-O-glucose and 19-O-glucose of the steviol glycoside and having 90% or greater sequence identity to the amino acid sequence set forth in SEQ ID NO:11;', '(ii) a second polypeptide capable of catalyzing beta 1,2 glycosylation of the C2′ of the 13-O-glucose, 19-O-glucose, or both 13-O-glucose and 19-O-glucose and having 70% or greater sequence identity to the amino acid sequence set forth in SEQ ID NO:17 or 18;', '(iii) a polypeptide capable of capable of catalyzing glycosylation of steviol or the steviol glycoside at its C-13 hydroxyl group and having 55% or greater sequence identity to the amino acid sequence set forth in SEQ ID NO:7; and/or', '(iv) a polypeptide capable of capable of beta 1,3 glycosylation of the C3′ of the 13-O-glucose, 19-O-glucose, or both 13-O-glucose and 19-O-glucose of the steviol glycoside having 50% or greater sequence identity to the amino acid sequence set forth in SEQ ID NO:9;, '(a) adding one or more of 'wherein at least one of the polypeptides is a recombinant polypeptide; and', 'and plant-derived- or synthetic steviol or steviol glycosides to a reaction mixture;'}(b) synthesizing steviol glycoside, glycosylated ent-kaurenol compound, and/or the glycosylated ent-kaurenoic acid compound in the reaction mixture.2. The method of claim 1 , further comprising:(c) recovering the steviol glycoside, glycosylated ent-kaurenol compound, and/or a glycosylated ent-kaurenoic acid compound alone or a composition comprising the steviol glycoside, glycosylated ent-kaurenol compound, and/or the glycosylated ent- ...

Подробнее
Номер записи: 67
28-01-2016 дата публикации

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

Номер: US20160024543A1
Автор: Heidtman Matthew Ian, McCoy John M., Merighi Massimo
Принадлежит:

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

Подробнее
Номер записи: 68
28-01-2021 дата публикации

Microbial production of steviol glycosides

Номер: US20210022373A1
Автор: Ajikumar Parayil Kumaran, Jason Donald, Krishna PATEL, Liwei Li, Ryan LIM, Ryan PHILIPPE, Swati Gupta
Принадлежит: Manus Bio Inc

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

Подробнее
Номер записи: 69
10-02-2022 дата публикации

METHOD FOR THE BIOSYNTHESIS OF DIOSMIN AND/OR HESPERIDIN IN A MICROORGANISM

Номер: US20220042061A1
Автор: JOUBERT LAETITIA, LE JEUNE ANDRÉ, PAUTHENIER CYRILLE, ROUSSEL CÉLIA, SCORNEC HÉLÈNE
Принадлежит:

The present invention relates to a recombinant microorganism which is modified to be capable of producing diosmin and hesperidin and to the use thereof for producing diosmin and/or hesperidin. 132-. (canceled)33. A recombinant microorganism comprising:a heterologous nucleic acid sequence coding for a flavanone 7-O-beta-D-glucosyltransferase (UGT) which is capable of adding a glucose in position 7 of hesperetin and/or diosmetin; anda heterologous nucleic acid sequence coding for a 6″-O-rhamnosyltransferase (RhaT) which is capable of transferring a rhamnose into position 6 of the glucose of hesperetin-7-O-glucoside and/or diosmetin-7-O-glucoside; anda heterologous nucleic acid sequence coding for a UDP-glucose 4,6-dehydratase/UDP-4-keto-6-deoxy-D-glucose 3,5-epimerase/UDP-4-keto-L-rhamnose-reductase (RHM) which is capable of producing UDP-rhamnose.34. The microorganism as claimed in claim 33 , in which the flavanone 7-O-beta-D-glucosyltransferase is selected from the group consisting of SEQ ID NOs: 113 claim 33 , 115 claim 33 , 91 claim 33 , 93 claim 33 , 95 claim 33 , 97 claim 33 , 99 and 101 and polypeptides comprising a sequence having at least 60% sequence identity with one of these sequences and having flavanone 7-O-beta-D-glucosyltransferase activity.35. The microorganism as claimed in claim 33 , in which the 6″-O-rhamnosyltransferase is selected from the group consisting of SEQ ID NOs: 103 and 105 and polypeptides comprising a sequence having at least 60% sequence identity with one of these sequences and having 6″-O-rhamnosyltransferase activity.36. The microorganism as claimed in claim 33 , in which the UDP-glucose 4 claim 33 ,6-dehydratase/UDP-4-keto-6-deoxy-D-glucose 3 claim 33 ,5-epimerase/UDP-4-keto-L-rhamnose-reductase is selected from the group consisting of SEQ ID NOs: 107 claim 33 , 109 and 111 and polypeptides comprising a sequence having at least 60% sequence identity with one of these sequences and having UDP-glucose 4 claim 33 ,6-dehydratase/UDP-4- ...

Подробнее
Номер записи: 70
24-01-2019 дата публикации

Transgenic pig which simultaneously expresses ho-1 gene and tnfr1-fc gene, and comprises knocked-out ggta1 gene, and use thereof

Номер: US20190024115A1
Автор: Bum Rae Cho, Byeong Chun Lee, Cu Rie Ahn, Eun Mi Lee, Geon A Kim, Hye Jin Hong, In Chang Hwang, Sang Hoon Lee, Su Cheong Yeom, Su Jin Kim
Принадлежит: Chong Kun Dang Corp, SEOUL NATIONAL UNIVERSITY R&DB FOUNDATION

The present invention relates to a transgenic pig in which an immune rejection response is suppressed during xenotransplantation, wherein a gene coding for heme oxygenase-1 (HO-1) and a gene coding for tumor necrosis factor receptor 1-Fc (TNFR1-Fc) are simultaneously expressed and a gene coding for α-1,3-galactosyltransferase (GGTA1) is knocked out; and a method for producing the same. The transgenic pig of the present invention, in which the genes coding for human HO-1 and TNFR1-Fc fusion protein are simultaneously expressed and the gene coding for GGTA1 is knocked out, may reduce oxidative stress during organ isolation and in vitro culture by antioxidative reaction, cytoprotective function, etc., and may also reduce a TNF-α-mediated inflammatory response in early transplantation by TNFR1-Fc expression. In addition, the transgenic pig may inhibit the maturation of dendritic cells and regulate the activation and proliferation of T-cells, thereby reducing an acute vascular rejection response to promote early engraftment of a transplanted organ. In addition, the transgenic pig can increase the viability of a transplanted organ by suppressing a hyper-acute immune rejection reaction caused by GGTA1. Accordingly, an organ, in which an immune rejection response is suppressed during xenotransplantation, can be produced using the transgenic pig.

Подробнее
Номер записи: 71
28-01-2021 дата публикации

COMPOSITIONS AND METHODS RELATING TO A MUTANT CLOSTRIDIUM DIFFICILE TOXIN

Номер: US20210024903A1
Автор: Anderson Annaliesa Sybil, Donald Robert G. K., Flint Michael James, Jansen Kathrin Ute, Kalyan Narender Kumar, Lotvin Jason Arnold, Moran Justin Keith, Ruppen Mark Edward, Sidhu Maninder K., Sun Weiqiang
Принадлежит:

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

Подробнее
Номер записи: 72
02-02-2017 дата публикации

FORMULATIONS COMPRISING POLYMERIC XYLOGLUCAN AS A CARRIER FOR AGRICULTURALLY BENEFICIAL AGENTS

Номер: US20170027167A1
Автор: Benyamino Romil, BERLIN Alex, Diano Audrey, Quinlan Jason
Принадлежит: NOVOZYMES BIOAG A/S

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

Подробнее
Номер записи: 73
02-02-2017 дата публикации

TREATMENT OF HYPERBILIRUBINEMIA

Номер: US20170028036A1
Автор: BORTOLUSSI GIULIA, COLLAUD FANNY, Mingozzi Federico, MURO ANDRÉS, RONZITTI GIUSEPPE
Принадлежит:

The invention relates to a nucleic acid sequence useful in the treatment of hyperbilirubinemia, in particular in the treatment of Crigler-Najjar syndrome. More particularly, the nucleic acid sequence of the present invention is a codon-optimized UGT1A1 coding sequence. 136-. (canceled)37. A nucleic acid sequence which is a codon optimized UGT1A1 coding sequence , wherein the optimized coding sequence has an increased GC content and/or has a decreased number of alternative open reading frames as compared to the wild-type coding sequence.38. The nucleic acid sequence according to claim 37 , comprising a nucleotide sequence at least 80% identical to SEQ ID NO:2 or SEQ ID NO:339. The nucleic acid sequence according to claim 38 , comprising the nucleotide sequence shown in SEQ ID NO:2 or SEQ ID NO:3.40. A nucleic acid construct comprising the nucleic acid sequence according to .41. The nucleic acid construct according to claim 40 , wherein said nucleic acid construct comprises said nucleic acid sequence operably linked to a promoter is an expression cassette.42. The nucleic acid construct according to claim 41 , wherein the promoter is a liver-specific promoter.43. The nucleic acid construct according to claim 41 , wherein the promoter is selected from the group consisting of the hAAT promoter claim 41 , alpha-1 antitrypsin promoter (hAAT) claim 41 , the transthyretin promoter claim 41 , the albumin promoter and the thyroxine-binding globulin (TBG) promoter.44. The nucleic acid construct according to claim 41 , said nucleic acid construct further comprising an intron.45. The nucleic acid construct according to claim 44 , wherein the intron is selected from the group consisting of a human beta globin b2 (or HBB2) intron claim 44 , a FIX intron and a chicken beta-globin intron.46. The nucleic acid construct according to claim 44 , wherein the intron is a modified intron with decreased or no alternative open reading frames (ARFs) claim 44 ,47. The nucleic acid construct ...

Подробнее
Номер записи: 74
29-01-2015 дата публикации

Immunization against clostridium difficile disease

Номер: US20150030612A1
Автор: Paul J. Giannasca, Thomas P. Monath, Wende Lei, William D. Thomas, Jr., Zhenxi Zhang
Принадлежит: Sanofi Pasteur Biologics LLC

The invention provides active and passive immunization methods for preventing and treating Clostridium difficile infection, which involve percutaneous administration of C. difficile toxin-neutralizing polyclonal immune globulin, C. difficile toxoids, or combinations thereof. Also provided by the invention are C. difficile toxoids, C. difficile toxin-neutralizing polyclonal immune globulin, and methods of identifying subjects that produce C. difficile toxin-neutralizing polyclonal immune globulin.

Подробнее
Номер записи: 75
04-02-2016 дата публикации

Toxoid, Compositions and Related Methods

Номер: US20160030542A1
Автор: Mark Shieh, Mike SOIKA
Принадлежит: Sanofi Pasteur Inc

Methods of producing purified clostridial toxin comprising tangential flow filtration, hydrophobic interaction chromatography and anion exchange chromatography are disclosed. These methods provide good yields of C. difficile toxin having a purity of about 90% or greater. Highly purified Clostridial toxins, toxoids (e.g., prepared by inactivating the toxin as disclosed herein) and compositions comprising these toxins and/or toxoids are also disclosed. Methods of using the purified toxins and/or toxoids for example, to elicit an immune response against Clostridium (e.g., C. difficile ) are also disclosed.

Подробнее
Номер записи: 76
01-02-2018 дата публикации

METHODS AND COMPOSITIONS FOR TREATMENT OF FORBES-CORI DISEASE

Номер: US20180028676A1
Автор: Armstrong Dustin D.
Принадлежит:

In certain embodiments, the present disclosure provides compositions and methods for treating Forbes-Cori Disease. 1171.-. (canceled)172. A method of treating Forbes-Cori disease in a subject in need thereof , comprising contacting the cell with a chimeric polypeptide comprising: (i) a mature acid alpha-glucosidase (GAA) polypeptide and (ii) an internalizing moiety that promotes delivery into cells;wherein the chimeric polypeptide has acid alpha-glucosidase activity, and wherein the chimeric polypeptide does not comprise a GAA precursor polypeptide of approximately 110 kilodaltons.173. The method of claim 172 , wherein the mature GAA polypeptide has a molecular weight of approximately 70-76 kilodaltons.174. The method of claim 172 , wherein the mature GAA polypeptide consists of an amino acid sequence selected from residues 122-782 of SEQ ID NO: 4 or residues 204-782 of SEQ ID NO: 5.175. The method of claim 172 , wherein the internalizing moiety promotes delivery of the chimeric polypeptide into cells.176. (canceled)177. (canceled)178. The method of claim 172 , wherein said chimeric polypeptide reduces cytoplasmic glycogen accumulation.179. The method of claim 172 , wherein the mature GAA polypeptide is glycosylated.180. The method of claim 172 , wherein the mature GAA polypeptide is not glycosylated.181. (canceled)182. The method of claim 172 , wherein the internalizing moiety comprises an antibody or antigen binding fragment.183. The method of claim 182 , wherein said antibody is a monoclonal antibody or fragment thereof claim 182 , orwherein said antibody is monoclonal antibody 3E10, or an antigen binding fragment thereof.184. (canceled)185. The method of claim 172 , wherein the internalizing moiety transits cellular membranes via an equilibrative nucleoside transporter claim 172 , orwherein the internalizing moiety transits cellular membranes via an equilibrative nucleoside transporter 2 (ENT2) transporter.186189.-. (canceled)190. The method of claim 182 , ...

Подробнее
Номер записи: 77
29-01-2015 дата публикации

DITERPENE PRODUCTION

Номер: US20150031868A1
Автор: BOER Viktor Marius, Kumar Manoj, LEHMANN Martin, Meijrink Bernard, SAGT CORNELIS MARIA JACOBUS, Trueheart Joshua, VAN DEN BERG Marco Alexander, Wu Liang, Zwart Jens Priscilla
Принадлежит:

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

Подробнее
Номер записи: 78
02-02-2017 дата публикации

METHOD FOR IMPROVING CROP PRODUCTIVITY

Номер: US20170029839A1
Автор: BLOMSTEDT Cecilia Karstin Maria, GAFF Donald Frederick, GRIFFITHS Cara, HAMILL John Diarmuid, ISLAM Sharmin, LE Ngoc Tuan, NEALE Alan Donald
Принадлежит:

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

Подробнее
Номер записи: 79
04-02-2016 дата публикации

Process for the production of hyaluronic acid in escherichia coli or bacillus megaterium

Номер: US20160032337A1
Автор: Alessandro Negro, Sonia Bisicchia, Vincenza Corsa
Принадлежит: Fidia Farmaceutici SpA

A method of producing hyaluronic acid (HA) in Escherichia coli and Bacillus megaterium through episomal plasmid vectors wherein the gene is under the control of strong promoter T7, preferably under the control of strong promoter T7 of bacteriophage T7, and a system for the selection of stable bacterial strains producing high levels of hyaluronic acid, are provided.

Подробнее
Номер записи: 80
31-01-2019 дата публикации

CREATION OF CHRYSANTHEMUM WITH BLUE FLOWER COLOR

Номер: US20190032066A1
Автор: Aida Ryutaro, HONGO Satoshi, Noda Naonobu, Sato Sanae, Tanaka Yoshikazu
Принадлежит:

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

Подробнее
Номер записи: 81
30-01-2020 дата публикации

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

Номер: US20200032308A1
Автор: Jennewein Stefan, Parschat Katja
Принадлежит:

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 ...

Подробнее
Номер записи: 82
04-02-2021 дата публикации

Microbial production of triterpenoids including mogrosides

Номер: US20210032669A1
Автор: Ajikumar Parayil Kumaran, Christine Nicole S. SANTOS, Michelle N. Goettge, Ryan PHILIPPE
Принадлежит: Manus Bio Inc

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.

Подробнее
Номер записи: 83
09-02-2017 дата публикации

MODIFIED HOST CELLS AND USES THEREOF

Номер: US20170035900A1
Автор: FERON Christiane Marie-Paule Simone Jeanne, KEMMLER Stefan J., KOWARIK Michael, QUEBATTE Julien L.
Принадлежит:

Described herein are modified host cells useful in the production of bioconjugates that can be used to vaccinate subjects against infection with . The genomes of the modified host cells described herein comprise genes that encode proteins involved in glyosylation of proteins as well as genes specific to the production of -specific antigens. 1102-. (canceled)103. A host cell comprising:{'i': 'Pseudomonas,', '(i) a nucleic acid that encodes a glycosyltransferase derived from an rfb cluster of'} (a) a polypeptide comprising a formyltransferase enzyme, wherein said nucleic acid encodes a protein having about or at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity or homology to SEQ ID NO:2, or wherein said nucleic acid encodes SEQ ID NO:2;', '(b) a wzy polymerase, wherein said nucleic acid encodes a protein having about or at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity or homology to SEQ ID NO:3, or wherein said nucleic acid encodes SEQ ID NO:3; and', {'i': 'Pseudomonas', '(c) a glycosyltransferase derived from an rfb cluster of , wherein said nucleic acid has been stably inserted into the genome of the host cell;'}], '(ii) a nucleic acid that encodes a polypeptide selected from the group consisting of(iii) a nucleic acid that encodes an oligosaccharyl transferase, and(iv) a nucleic acid that encodes a carrier protein comprising an N-glycosylation consensus, wherein if the polypeptide is (a) or (b), then the N-glycosylation consensus sequence is D/E-X-N-X-S/T and X is any amino acid except proline.104Pseudomonas aeruginosa. The host cell of claim 103 , wherein the glycotransferase is derived from an rfb cluster of serotype 6.105E. coli.. The host cell of claim 103 , wherein said host cell is106Pseudomonas. A method of producing a bioconjugate that comprises a antigen linked to a carrier protein claim 103 , said method comprising culturing the host cell of under conditions suitable for the production of proteins.107Pseudomonas. A bioconjugate produced ...

Подробнее
Номер записи: 84
09-02-2017 дата публикации

Processes for producing hydrocarbon products

Номер: US20170037320A1
Автор: Anna EL TAHCHY, James Robertson Petrie, Qing Liu, Surinder Singh, Thomas Vanhercke
Принадлежит: Commonwealth Scientific and Industrial Research Organization CSIRO

The present invention relates to processes for producing industrial products such as hydrocarbon products from non-polar lipids in a vegetative plant part. Preferred industrial products include alkyl esters which may be blended with petroleum based fuels.

Подробнее
Номер записи: 85
08-02-2018 дата публикации

NON-CALORIC SWEETENER

Номер: US20180037600A1
Автор: CHATURVEDULA VENKATA SAI PRAKASH, Mao Guohong, Yu Xiaodan
Принадлежит: Conagen Inc.

Disclosed is a steviol glycoside referred to as rebaudioside D2. Rebaudioside D2 has five β-D-glucosyl units connected to the aglycone steviol. Also disclosed are methods for producing rebaudioside D2, a UDP-glycosyltransferase fusion enzyme, and methods for producing rebaudioside D and rebaudioside E. 132-. (canceled)33. A method for synthesizing rebaudioside D , the method comprising:preparing a reaction mixture comprising rebaudioside A; a substrate selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose); and a UDP-glycosyltransferase selected from the group consisting of a uridine diphospho glycosyltransferase and a UDP-glycosyltransferase fusion enzyme comprising a uridine diphospho glycosyltransferase domain coupled to a sucrose synthase domain; and incubating the reaction mixture for a sufficient time to produce rebaudioside D, wherein a glucose is covalently coupled to the rebaudioside A to produce rebaudioside D.34Oryza sativa. The method of claim 33 , wherein the UDP-glycosyltransferase is a uridine diphospho glycosyltransferase EUGT11.35. The method of further comprising adding a sucrose synthase to the reaction mixture.36ArabidopsisCoffeaStevia. The method of claim 35 , wherein the sucrose synthase is selected from the group consisting of an sucrose synthase 1; a sucrose synthase 1 and a sucrose synthase 1.37Arabidopsis thaliana. The method of claim 35 , wherein the sucrose synthase is an sucrose synthase 1.38. The method of claim 33 , wherein the UDP-glycosyltransferase is a UDP-glycosyltransferase fusion enzyme comprising a uridine diphospho glycosyltransferase domain coupled to a sucrose synthase domain.39. The method of claim 38 , wherein the UDP-glycosyltransferase fusion enzyme comprises an amino acid sequence having about 90% sequence identity to SEQ ID NO:5.40Oryza sativa. The method of claim 33 , wherein the uridine diphospho glycosyltransferase domain is an uridine diphospho ...

Подробнее
Номер записи: 86
08-02-2018 дата публикации

BETA-HEXOSYL-TRANSFERASES AND USES THEREOF

Номер: US20180037873A1
Автор: Azcarate-Peril Maria, BRUNO-BARCENA JOSE M., DAGHER Suzanne
Принадлежит:

This invention relates generally to the discovery of novel recombinant forms of β-hexosyl-transferases (BHT) and uses thereof to produce galacto-oligosaccharides (GOS) or as food additives. 1. An isolated DNA encoding a recombinant β-hexosyl-transferase (rBHT) protein having the amino acid sequence set forth in SEQ ID NO: 4 , 6 , 8 , 10 , 12 , 14 , 16 , 18 or 20.2. The isolated DNA encoding the recombinant β-hexosyl-transferase (rBHT) protein of having the amino acid sequence set forth in SEQ ID NO: 12 or 14.3. The isolated DNA encoding the recombinant β-hexosyl-transferase (rBHT) protein of claim 1 , wherein the DNA has at least 97% sequence identity with the nucleic acid sequence set forth in SEQ ID NO: 3 claim 1 , 5 claim 1 , 7 claim 1 , 9 claim 1 , 11 claim 1 , 13 claim 1 , 15 claim 1 , 17 or 19.4. The isolated DNA encoding the recombinant β-hexosyl-transferase (rBHT) protein of claim 3 , wherein the DNA has at least 97% sequence identity with the nucleic acid sequence set forth in SEQ ID NO: 13 or 15.5. An enzymatically active recombinant β-hexosyl-transferase (rBHT) protein wherein the protein has the amino acid sequence set forth in SEQ ID NO: 4 claim 3 , 6 claim 3 , 8 claim 3 , 10 claim 3 , 12 claim 3 , 14 claim 3 , 16 claim 3 , 18 or 20.6. The enzymatically active rBHT protein of claim 5 , wherein the protein is membrane bound.7. The enzymatically active rBHT protein of claim 5 , wherein the protein is a soluble enzyme.8. A method for producing enzymatically active recombinant β-hexosyl-transferase (rBHT) protein in a cultured eukaryotic host cell which comprises transforming the eukaryotic host cell with a plasmid under the control of a suitable promotor wherein the plasmid contains an isolated DNA encoding a rBHT protein having the amino acid sequence set forth in SEQ ID NO: 4 claim 5 , 6 claim 5 , 8 claim 5 , 10 claim 5 , 12 claim 5 , 14 claim 5 , 16 claim 5 , 18 or 20 wherein the promoter is operably linked to the isolated DNA claim 5 , and culturing ...

Подробнее
Номер записи: 87
08-02-2018 дата публикации

Methods for Producing Diterpenes

Номер: US20180037912A1
Автор: Andersen-Ranberg Johan, Bohlmann Carl Jorg, Hamberger Bjorn, Lindberg Moller Birger, Nielsen Morten Thrane, Zerbe Philipp
Принадлежит:

The present invention discloses that by combining different di TPS enzymes of class I and class II different diterpenes may be produced including diterpenes not identified in nature. Surprisingly it is revealed that a di TPS enzyme of class I of one species may be combined with a di TPS enzyme of class II from a different species, resulting in a high diversity of diterpenes, which can be produced. 1. A method of producing a terpene , comprising: i. A heterologous nucleic acid encoding a diTPS of class II,', 'with the proviso that the diTPS of class II and the diTPS of class I are not from same species; and with the proviso that when the diTPS of class II is SsLPPS then the diTPS of class I is not CfTPS3, CfTPS4 or EpTPS8 and when the diTPS of class I is EpTPS8, then the diTPS of class II is not CfTPS2 or SsLPPS;', 'ii. A heterologous nucleic acid encoding a diTPS of class I,'}], '(a) providing a host organism comprising'}(b) incubating the host organism in the presence of geranylgeranyl pyrophosphate (GGPP) under conditions allowing growth of the host organism; andc) Optionally isolating diterpene from the host organism.2. The method of claim 1 , wherein the diterpene is a C-molecule containing a decalin core and up to 3 oxygen molecules.4. The method of claim 3 , wherein the diterpene is a C-molecule containing a cores structure of formula I claim 3 , II claim 3 , III claim 3 , IV claim 3 , V claim 3 , VI claim 3 , IX or X substituted at one or more positions by one or more groups comprising:(a) alkyl, wherein the alkyl is linear or branched;(b) alkenyl; and(c) hydroxyl.5. The method of claim 1 , wherein the diterpene is a C-molecule containing a decalin substituted at the 10 position with C-alkenyl chain claim 1 , a hydroxyl claim 1 , a methyl group and/or ═C.6. The method of claim 1 , wherein the diterpene is a C-molecule consisting of 20 carbon atoms claim 1 , with up to three oxygen atoms and hydrogen atoms claim 1 , wherein the molecule contains a core ...

Подробнее
Номер записи: 88
08-02-2018 дата публикации

PROTECTING GROUP CHEMISTRY FOR CLEAN, REDUCTANT-FREE DYEING

Номер: US20180037917A1
Автор: BHAKTA Shyam Pravin, CERVANTES Bernardo, DUEBER John Eugene, FONG, HSU Tammy Melody, III Arthur Muir, Johnson, Jr. Terry Don, LATIMER Luke Nathaniel, PRATHURI Ramya Lakshmi, RUSS Zachary Nicholas
Принадлежит: The Regents of the University of California

The present disclosure relates to the biosynthesis of indigoid dye precursors and their conversion to indigoid dyes. Specifically, the present disclosure relates to methods of using polypeptides to produce indigoid dye precursors from indole feed compounds, and the use of the indigoid dye precursors to produce indigoid dyes. 2. The method of claim 1 , wherein the host cell is a bacterial cell.3E. coliCorynebacterium glutamicum.. The method of claim 2 , wherein the host cell is selected from the group consisting of and4. The method of claim 1 , wherein the host cell is a eukaryotic cell.5. The method of claim 4 , wherein the host cell is a yeast.6Saccharomyces cerevisiae, Pichia pastoris, Kluyveromyces lactis, Kluyveromyces marxianus,Schizosaccharomyces pombe.. The method of claim 4 , wherein the host cell is selected from the group consisting of and7. The method of claim 1 , wherein the polypeptide with oxygenase activity comprises SEQ ID NO: 1 claim 1 , SEQ ID NO: 2 claim 1 , SEQ ID NO: 21 claim 1 , SEQ ID NO: 22 claim 1 , SEQ ID NO: 23 claim 1 , or SEQ ID NO: 24 claim 1 , or the amino acid sequence of any homologs thereof.8. The method of claim 1 , wherein the polypeptide with glycosyltransferase activity comprises SEQ ID NO: 3 claim 1 , SEQ ID NO: 4 claim 1 , SEQ ID NO: 5 claim 1 , SEQ ID NO: 6 claim 1 , SEQ ID NO: 7 claim 1 , SEQ ID NO: 8 claim 1 , SEQ ID NO: 15 claim 1 , SEQ ID NO: 16 claim 1 , SEQ ID NO: 17 claim 1 , SEQ ID NO: 18 claim 1 , SEQ ID NO: 19 claim 1 , or SEQ ID NO: 20 claim 1 , or the amino acid sequence of any homologs thereof.9. The method of claim 1 , wherein the polypeptide with sulfotransferase activity comprises SEQ ID NO: 9 claim 1 , SEQ ID NO: 10 claim 1 , SEQ ID NO:11, SEQ ID NO: 12, SEQ ID NO: 13, or SEQ ID NO: 14, or the amino acid sequence of any homologs thereof.10. The method of claim 7 , wherein the homolog of a polypeptide with oxygenase activity comprises an amino acid sequence at least 90% claim 7 , at least 95% claim 7 , or at ...

Подробнее
Номер записи: 89
07-02-2019 дата публикации

TRANSGENIC SILKWORM HAVING MAMMALIAN-TYPE SUGAR CHAIN ATTACHED THERETO

Номер: US20190037820A1
Автор: Fujiyama Kazuhito, MISAKI Ryo, Sezutsu Hideki, Tatematsu Ken-ichiro
Принадлежит:

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

Подробнее
Номер записи: 90
12-02-2015 дата публикации

VACCINES AGAINST CLOSTRIDIUM DIFFICILE COMPRISING RECOMBINANT TOXINS

Номер: US20150044250A1
Автор: Bodmer Jean-Luc, Caro-Aguilar Ivette, Gentile Marie-Pierre, Goerke Aaron Rudy, Heijden-Liefkens Karin Huberdina Antonia van der, Heinrichs Jon Henry, Horton Melanie S., Miezeiewski Matthew Ryan, Secore Susan Lynn, Skinner Julie M., Sondermeijer Paulus Jacobus Antonius, Subramanian Shyamsundar, Wang Su, Xie Jinfu, Xoconostle Rachel Flora, Zorman Julie K.
Принадлежит:

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

Подробнее
Номер записи: 91
24-02-2022 дата публикации

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

Номер: US20220056497A1
Автор: Heidtman Matthew Ian, McCoy John M., Merighi Massimo
Принадлежит:

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 ...

Подробнее
Номер записи: 92
07-02-2019 дата публикации

GLUCOSYLTRANSFERASE ENZYMES FOR PRODUCTION OF GLUCAN POLYMERS

Номер: US20190040432A1
Автор: Brun Yefim, HE HONGXIAN, PAYNE MARK S., Scholz Thomas
Принадлежит: E I DU PONT DE NEMOURS AND COMPANY

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 ...

Подробнее
Номер записи: 93
06-02-2020 дата публикации

O-Mannosyltransferase Deficient Filamentous Fungal Cells and Methods of Use Thereof

Номер: US20200040314A1
Автор: Hiltunen Jukka, Huuskonen Anne, Natunen Jari, Ostermeier Christian, Saloheimo Markku, Sommer Benjamin Patrick, Wahl Ramon
Принадлежит: GLYKOS FINLAND OY

The present disclosure relates to compositions and methods useful for the production of heterologous proteins with reduced O-mannosylation in filamentous fungal cells, such as cells. More specifically, the invention provides a PMT-deficient filamentous fungal cell comprising a) at least a first mutation that reduces an endogenous protease activity compared to a parental filamentous fungal cell which does not have said first mutation, and, b) at least a second mutation in a PMT gene that reduces endogenous O-mannosyltransferase activity compared to a parental filamentous fungal cell which does not have said second mutation, wherein said filamentous fungal cell is selected from the group consisting of or cell. 1. A PMT-deficient filamentous fungal cell comprisinga) a first mutation in a gene encoding an endogenous protease that reduces or eliminates an endogenous protease activity as compared to a parental filamentous fungal cell which does not have said first mutation, said endogenous protease being selected among aspartic proteases, trypsin-like serine proteases, subtilisin proteases, glutamic proteases and sedolisin proteases, andb) a second mutation in a PMT gene that reduces endogenous O-mannosyltransferase activity compared to a parental filamentous fungal cell which does not have said second mutation,{'i': Trichoderma, Neurospora, Myceliophthora', 'Chrysosporium, 'wherein said filamentous fungal cell is selected from the group consisting of and cell.'}2. The PMT-deficient filamentous fungal cell of claim 1 , wherein said second mutation that reduces the endogenous O-mannosyltransferase activity is a deletion or a disruption of a PMT gene encoding an endogenous protein O-mannosyltransferase activity.3. The PMT-deficient filamentous fungal cell of claim 1 , wherein said second mutation in a PMT gene is a mutation in either:a) PMT1 gene comprising the polynucleotide of SEQ ID NO:1,{'i': 'T. reesei', 'b) a homologous gene of a PMT1 gene, which homologous gene is ...

Подробнее
Номер записи: 94
18-02-2016 дата публикации

GLUCURONOSYLTRANSFERASE, GENE ENCODING SAME AND USE THEREOF

Номер: US20160046912A1
Автор: MURANAKA Toshiya, Ohyama Kiyoshi, SAITO Kazuki, Seki Hikaru
Принадлежит: RIKEN

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

Подробнее
Номер записи: 95
18-02-2016 дата публикации

Enhanced itaconic acid production in aspergillus with increased laea expression

Номер: US20160046967A1
Автор: Scott E. Baker, Ziyu Dai
Принадлежит: Battelle Memorial Institute Inc

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

Подробнее
Номер записи: 96
16-02-2017 дата публикации

BIOLOGICAL PLATFORM FOR PRODUCTION OF COMMODITY CHEMICALS

Номер: US20170044583A1
Автор: FAN Zhiliang, HILDEBRAND Amanda
Принадлежит:

The present disclosure generally relates to biological platforms for the conversion of cellulosic biomass into fuels and chemicals. More specifically, the present disclosure relates to the conversion of cellulosic materials into sugar acids or their salts, which may then be used to produce commodity chemicals. In one aspect, the present disclosure relates to a recombinant host cell including: reduced activity of one or more polypeptides having P-glucosidase activity as compared to a corresponding wild type cell, where each of said one or more polypeptides are encoded by a gene that has at least 80% sequence identity to a gene. 1. A recombinant host cell comprising:a) reduced activity of one or more polypeptides having β-glucosidase activity as compared to a corresponding wild type cell, wherein each of said one or more polypeptides are encoded by a gene that has at least 80% sequence identity to a gene selected from the group consisting of NCU00130, NCU04952, NCU05577, NCU07487, NCU08755, and NCU03641 genes;b) reduced activity of a polypeptide having cellobionate phosphorylase activity as compared to a corresponding wild type cell, wherein said polypeptide is encoded by a gene that has at least 80% sequence identity to NCU09425 (NdvB);c) reduced activity of a polypeptide encoded by a gene that has at least 80% sequence identity to NCU08807 (CRE-1) as compared to a corresponding wild type cell; andd) reduced activity of a polypeptide encoded by a gene that has at least 80% sequence identity to NCU09333 (ACE-1) as compared to a corresponding wild type host cell.2. The host cell of claim 1 , wherein said host cell is a fungal cell.3Neurospora crassa.. The host cell of claim 2 , wherein said fungal cell is4. The host cell of any one of - claim 2 , wherein reduced activity of one or more of the polypeptides having β-glucosidase activity is due to a genetic mutation.5. The host cell of claim 4 , wherein a genetic mutation is present in two or more claim 4 , three or more ...

Подробнее
Номер записи: 97
15-02-2018 дата публикации

Polypeptides Having Glucuronyl Esterase Activity and Polynucleotides Encoding Same

Номер: US20180044646A1
Автор: Anders Viksoe-Nielsen, Johan Börjesson, Kristian Krogh, Nikolaj Spodsberg
Принадлежит: Novozymes AS

The present invention relates to isolated polypeptides having glucuronyl esterase activity, catalytic domains and polynucleotides encoding the polypeptides or catalytic domains. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides or catalytic domains.

Подробнее
Номер записи: 98
19-02-2015 дата публикации

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

Номер: US20150051381A1
Автор: Byung-Kwon Choi, Juergen Hermann Nett, Piotr Bobrowicz, Robert C. Davidson, Stefan Wildt, Stephen R. Hamilton, Tillman U. Gerngross
Принадлежит: GLYCOFI, INC.

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

Подробнее
Номер записи: 99
03-03-2022 дата публикации

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

Номер: US20220064608A1
Автор: Chen Yujie
Принадлежит:

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

Подробнее
Номер записи: 100