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 20638. Отображено 100.
02-02-2012 дата публикации

Preparation of adipic acid

Номер: US20120028320A1
Автор: Axel Christoph Trefzer, Martin SCHÜRMANN, Petronella Catharina Raemakers-Franken, Stefaan Marie Andre De Wildeman
Принадлежит: DSM IP ASSETS BV

The invention relates to a method for preparing adipic acid, comprising converting alpha-ketoglutaric acid (AKG) into alpha-ketoadipic acid (AKA), converting alpha-ketoadipic acid into alpha-ketopimelic acid (AKP), converting alpha-ketopimelic acid into 5-formylpentanoic acid (5-FVA), and converting 5-formylpentanoic acid into adipic acid, wherein at least one of these conversions is carried out using a heterologous biocatalyst.The invention further relates to a heterologous cell, comprising one or more heterologous nucleic acid sequences encoding one or more heterologous enzymes capable of catalysing at least one reaction step in said method.

Подробнее
Номер записи: 1
02-02-2012 дата публикации

Yeast organism producing isobutanol at a high yield

Номер: US20120028323A1
Автор: Aristos Aristidou, Catherine Asleson Dundon, Christopher Smith, Jun URANO, Peter Meinhold, Reid M. Renny FELDMAN, Uvini GUNAWARDENA
Принадлежит: Gevo Inc

The present invention provides recombinant microorganisms comprising an isobutanol producing metabolic pathway and methods of using said recombinant microorganisms to produce isobutanol. In various aspects of the invention, the recombinant microorganisms may comprise a modification resulting in the reduction of pyruvate decarboxylase and/or glycerol-3-phosphate dehydrogenase activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

Подробнее
Номер записи: 2
09-02-2012 дата публикации

Method for observing gad67-positive cell in transgenic animal

Номер: US20120036588A1
Автор: Yuchio Yanagawa
Принадлежит: JAPAN SCIENCE AND TECHNOLOGY AGENCY

A method for observing glutamate decarboxylase 67-positive cells in a transgenic mouse includes providing the transgenic mouse in which a nucleic acid sequence encoding green fluorescent protein is inserted in a frame within exon 1 of an endogenous glutamate decarboxylase 67 gene, whereby the transgenic mouse is capable of a functional expression of the green fluorescent protein in place of the endogenous glutamate decarboxylase 67 gene. Then, the glutamate decarboxylase 67-positive cells which are specifically visualized by the expression of the green fluorescent protein can be observed. The method may further include analyzing a function or morphology of the GABAergic neurons based on the observation of the glutamate decarboxylase 67-positive cells where the glutamate decarboxylase 67-positive cells are specifically visualized corresponding to a cell distribution of GABAergic neurons.

Подробнее
Номер записи: 3
16-02-2012 дата публикации

Novel aldolase and production process of substituted alpha-keto acids

Номер: US20120040416A1
Автор: Kunihiko Watanabe, Masakazu Sugiyama, Shigeru Kawahara
Принадлежит: Ajinomoto Co Inc

4-(Indol-3-ylmethyl)-4-hydroxy-2-oxoglutarate, which is useful as an intermediate in the synthesis of monatin, may be synthesized from indole pyruvic acid and pyruvic acid (and/or oxaloacetic acid) by using a novel aldolase derived from the genus Pseudomonas, Erwinia, Flavobacterium , or Xanthomonas.

Подробнее
Номер записи: 4
08-03-2012 дата публикации

Homolactic Fermentation from Pentose

Номер: US20120058528A1
Автор: Akihiko Kondo, Hideo Noda, Kenji Okano
Принадлежит: Bio Energy Corp, Kansai Chemical Engineering Co Ltd

Provided is a lactic acid bacterium capable of homolactic fermentation using a pentose as a substrate, the lactic acid bacterium utilizing a pentose, and in which a phosphoketolase pathway is blocked and a pentose phosphate pathway is activated. Also provided is a method for producing lactic acid from a pentose using the lactic acid bacterium and a method for preparing the lactic acid bacterium.

Подробнее
Номер записи: 5
22-03-2012 дата публикации

Dbc1, a novel native inhibitor of the anti-aging protein sirt1

Номер: US20120070843A1
Автор: Wei Gu
Принадлежит: Columbia University of New York

A novel complex is identified between the NAD-dependent deacetylase, SIRT1 and its novel inhibitor, DBC1. Provided herein are methods to indentify a compound that inhibits the complexation between SIRT1 and DBC1. Exemplary methods comprise contacting either the complexation between DBC1 and SIRT1 with an agent being tested for its ability to inhibit the complexation between SIRT1 and DBC1. Also, provided are methods to identify a compound that increases the complexation between SIRT1 and DBC1. Exemplary methods comprise contacting either the complexation between DBC1 and SIRT1 with an agent being tested for its ability to increase the complexation between SIRT1 and DBC1. Further, methods are provided to increase or decrease SIRT1 activity by contacting the complexation between SIRT1 and DBC1 with a peptide that either decreases or increases the complexation between SIRT1 and DBC1. Further, methods are provided for the treatment of patients suffering from diseases including metabolic diseases including obesity and diabetes, and neurodegenerative disorders including Alzheimer's disease and Huntington's disease using compounds that inhibit the complexation between SIRT1 and DBC1.

Подробнее
Номер записи: 6
19-04-2012 дата публикации

Class ii human histone deacetylases, and uses related thereto

Номер: US20120094862A1
Автор: Christian A. Hassig, Christina M. Grozinger, Stuart L. Schreiber
Принадлежит: Harvard College

The invention provides histone deacetylase class II nucleic acids and polypeptides, methods and reagents for their use, and related compounds including small molecule libraries containing class II histone deacetylase inhibitors.

Подробнее
Номер записи: 7
07-06-2012 дата публикации

Recombinant Colwellia Psychrerythraea Alkaline Phosphatase and Uses Thereof

Номер: US20120142061A1
Автор: Christopher J. Kubu, Jeannine Muller-Greven, Marc A. Post
Принадлежит: Affymetrix Inc

A heat labile alkaline phosphatase enzyme and methods of using the same and kits including the same are disclosed. Specifically, a nucleotide sequence of, peptide sequence of, methods of using, and kits comprising, a heat labile alkaline phosphatase isolated from Colwellia psychrerythraea are provided. Methods of over-expression and purification of the recombinant alkaline phosphatase and mutants thereof are also disclosed. Methods of over-expressing and purifying commercially useful quantities of active recombinant heat labile alkaline phosphatase fusion enzymes from C. psychrerythraea, wherein the fusion enzymes comprise one or more heterologous leader sequences are disclosed. The disclosed C. psychrerythraea heat labile alkaline phosphatase has properties similar to shrimp alkaline phosphatase and can be substituted for shrimp alkaline phosphatase in assays involving the same.

Подробнее
Номер записи: 8
21-06-2012 дата публикации

Preparation of alpha-ketopimelic acid

Номер: US20120156737A1
Автор: Axel Christoph Trefzer, Martin SCHÜRMANN, Petronella Catharina Raemakers-Franken, Stefaan Marie Andre De Wildeman
Принадлежит: DSM IP ASSETS BV

The invention relates to a method for preparing alpha-ketopimelic acid, comprising converting alpha-ketoglutaric acid into alpha-ketoadipic acid and converting alpha-ketoadipic acid into alpha-ketopimelic acid, wherein at least one of these conversions is carried out using a heterologous biocatalyst. The invention further relates to a heterologous cell, comprising one or more heterologous nucleic acid sequences encoding one or more heterologous enzymes capable of catalysing at least one reaction step in the preparation of alpha-ketopimelic acid from alpha-ketoglutaric acid.

Подробнее
Номер записи: 9
28-06-2012 дата публикации

Succinic acid production in a eukaryotic cell

Номер: US20120165569A1
Автор: Cornelis Maria Jacobus Sagt, Liang Wu, Rene Verwaal, Robbertus Antonius Damveld
Принадлежит: DSM IP ASSETS BV

The present invention relates to a recombinant eukaryotic cell selected from a yeast of a filamentous fungus comprising a nucleotide sequence encoding a NAD(H)-dependent fumarate reductase that catalyses the conversion of fumaric acid to succinic acid. The invention further relates to a process for the production of succinic acid wherein the eukaryotic cell according to the present invention is used.

Подробнее
Номер записи: 10
12-07-2012 дата публикации

Mutated acetohydroxyacid synthase genes in brassica

Номер: US20120178628A1
Автор: Christian Schopke, Greg F. W. Gocal, Keith Walker, Peter R. Beetham
Принадлежит: Individual

Provided are mutated acetohydroxyacid synthase (AHAS) nucleic acids and the proteins encoded by the mutated nucleic acids. Also provided are canola plants, cells, and seeds comprising the mutated genes.

Подробнее
Номер записи: 11
02-08-2012 дата публикации

Novel Sesquiterpene Synthase Gene and Protein

Номер: US20120196340A1
Автор: Bryan Greenhagen, Joseph Chappell
Принадлежит: University of Kentucky Research Foundation

The invention relates to sesquiterpene synthases and methods for their production and use. Particularly, the invention provides nucleic acids comprising the nucleotide sequence of citrus valencene synthase (CVS) which codes for at least one CVS. The invention further provides nucleic acids comprising the nucleotide sequence coding for amino acid residues forming the tier 1 and tier 2 domains of CVS. The invention also provides for methods of making and using the nucleic acids and amino acids of the current invention.

Подробнее
Номер записи: 12
16-08-2012 дата публикации

Compositions and methods for promoting neuronal outgrowth

Номер: US20120207732A1
Автор: A. M. Vecchione, Elliott A. Gruskin, Gargi Roy, Jennifer F. Iaci, Sarah Kasperbauer
Принадлежит: Acorda Therapeutics Inc

Neural outgrowth in the central nervous system is achieved by administering chondroitinase AC and/or chondroitinase B to degrade chondroitin sulfate proteoglycans that inhibit or contribute to the inhibition of nervous tissue regeneration.

Подробнее
Номер записи: 13
23-08-2012 дата публикации

Plant biochemical systems and uses thereof

Номер: US20120214239A1
Автор: Katherine Warpeha, Lon Kaufman
Принадлежит: University of Illinois

Plants and plant cells including an isolated nucleic acid encoding for prephenate dehydratase, wherein the nucleic acid is operably linked to a promoter such that prephenate dehydratase is expressed at sufficient levels to protect the plant cell from damage from an abiotic or biotic stressor. The abiotic or biotic stressor may include UV radiation, cold, drought, heat, salt, hormones, fungi, bacteria, arthropods, worms and products of biotic organisms.

Подробнее
Номер записи: 14
30-08-2012 дата публикации

Recombinant Halohydrin Dehalogenase Polypeptides

Номер: US20120220002A1
Автор: Erik de Vries, Erika Segraves, Kheng Lin Tan, Louis Clark, Scott McVicar, Shiwei Song
Принадлежит: Codexis Inc

The present disclosure provides engineered halohydrin dehalogenase (HHDH) polypeptides having improved enzyme properties as compared to the wild-type HHDH enzyme HheC and other reference engineered HHDH polypeptides. Also provided are polynucleotides encoding the engineered HHDH enzymes, host cells capable of expressing the engineered HHDH enzymes, and methods of using the engineered HHDH enzymes to synthesize a variety of chiral compounds including chiral epoxides and chiral alcohols.

Подробнее
Номер записи: 15
06-09-2012 дата публикации

Method for the Production of Very Long Chain Fatty Acids (VLCFA) by Fermentation with a Recombinant Yarrowia SP

Номер: US20120226059A1
Автор: Brice Bourdenx, Jean-Denis Faure, Jean-Marc Nicaud, Ramdane Haddouche
Принадлежит: Institut National de la Recherche Agronomique INRA

The present invention concerns a method for the production of Very Long Chain Fatty Acids (VLCFA) by fermentation, comprising culturing a recombinant strain of a Yarrowia sp. comprising a heterologous gene coding for a hydroxyacyl-CoA dehydratase, under control of regulatory elements allowing expression of the said heterologous gene in the said Yarrowia sp. The invention also concerns the recombinant Yarrowia sp.

Подробнее
Номер записи: 16
20-09-2012 дата публикации

Microorganisms and methods for the biosynthesis of fumarate, malate, and acrylate

Номер: US20120237990A1
Автор: Anthony P. Burgard, Mark J. Burk, Priti Pharkya
Принадлежит: Genomatica Inc

A non-naturally occurring eukaryotic or prokaryotic organism includes one or more gene disruptions occurring in genes encoding enzymes imparting increased fumarate, malate or acrylate production in the organism when the gene disruption reduces an activity of the enzyme. The one or more gene disruptions confers increased production of acrylate onto the organism. Organisms that produce acrylate have an acrylate pathway that at least one exogenous nucleic acid encoding an acrylate pathway enzyme expressed in a sufficient amount to produce acrylate, the acrylate pathway comprising a decarboxylase. Methods of producing fumarate, malate or acrylate include culturing these organisms.

Подробнее
Номер записи: 17
25-10-2012 дата публикации

Methods of developing terpene synthase variants

Номер: US20120270260A1
Автор: Andrew MAIN, Lan Xu, Lishan Zhao, Patrick WESTFALL
Принадлежит: Amyris Inc

The present disclosure relates to methods of developing terpene synthase variants through engineered host cells. Particularly, the disclosure provides methods of developing terpene synthase variants with improved in vivo performance that are useful in the commercial production of terpene products. Further encompassed in the present disclosure are superior terpene synthase variants and host cells comprising such terpene synthase variants.

Подробнее
Номер записи: 18
17-01-2013 дата публикации

Nucleic acid construct comprising pyripyropene biosynthetic gene cluster and marker gene

Номер: US20130017581A1
Автор: Hiroyuki Anzai, Kentaro Yamamoto, Koichiro Murashima, Kouji Yanai, Naomi Sumida, Sato Aihara
Принадлежит: Individual

There is provided a nucleic acid construct comprising a pyripyropene biosynthetic gene cluster and a marker gene. The nucleic acid construct according to the present invention provides an inexpensive and highly productive method for producing pyripyropene.

Подробнее
Номер записи: 19
31-01-2013 дата публикации

Recombinant ethanologenic bacteria

Номер: US20130029393A1
Автор: Andrew MacDonald, David Nunn
Принадлежит: BP Corp North America Inc

The invention provides recombinant ethanologenic bacteria, methods of making the bacteria and methods of producing ethanol using the bacteria.

Подробнее
Номер записи: 20
21-02-2013 дата публикации

Isoprene synthase variants for improved production of isoprene

Номер: US20130045891A1
Автор: Christopher L. Rife, David A. Estell, Derek H. Wells, Jeffrey V. Miller, Zachary Q. Beck
Принадлежит: DANISCO US INC

The present invention provides methods and compositions comprising at least one isoprene synthase enzyme with improved specific productivity. In particular, the present invention provides variant plant isoprene synthases for increased isoprene production in host cells.

Подробнее
Номер записи: 21
07-03-2013 дата публикации

Novel amidase, gene for the same, vector, transformant, and method for production of optically active carboxylic acid amide and optically active carboxylic acid by using any one of those items

Номер: US20130059348A1
Автор: Daisuke Moriyama, Masutoshi Nojiri, Naoaki Taoka, Tozo Nishiyama
Принадлежит: Kaneka Corp

The present invention has its object to provide a novel polypeptide having amidase activity to selectively hydrolyze S-enantiomer in racemic nipecotamide, a DNA encoding the polypeptide, a vector containing the DNA, a transformant transformed with the vector, and a method for producing an optically active carboxylic acid amide and an optically active carboxylic acid in which a racemic carboxylic acid amide is hydrolyzed with the polypeptide or the transformant.

Подробнее
Номер записи: 22
21-03-2013 дата публикации

Chimeric isoprenoid synthases and uses thereof

Номер: US20130071877A1
Автор: Back Kyoungwhan, Chappell Joseph
Принадлежит:

Provided is a chimeric isoprenoid synthase polypeptide including a first domain from a first isoprenoid synthase joined to a second domain from a second, heterologous, isoprenoid synthase, whereby the chimeric isoprenoid synthase is capable of catalyzing the production of isoprenoid reaction products that are not produced in the absence of the second domain of the second, heterologous, isoprenoid synthase. Also provided is a chimeric isoprenoid synthase polypeptide including an asymmetrically positioned heterologous domain, whereby the chimeric isoprenoid synthase is capable of catalyzing the production of isoprenoid reaction products that are not produced when the domain is positioned at its naturally-occurring site in the isoprenoid synthase polypeptide. 1. A method for producing a chimeric isoprenoid synthase polypeptide , comprising:(a) providing a cell comprising DNA encoding the chimeric isoprenoid synthase polypeptide, wherein the encoded synthase comprises a first sesquiterpene isoprenoid synthase polypeptide domain joined to a second, different, sesquiterpene isoprenoid synthase polypeptide domain such that the chimeric isoprenoid synthase polypeptide produces one or more isoprenoid products that are not produced in the absence of the second domain of the second synthase;(b) culturing said cell under conditions for expressing the DNA to produce the chimeric isoprenoid synthase; and(c) recovering the chimeric isoprenoid synthase.2. The method of claim 1 , wherein the encoded chimeric isoprenoid synthase comprises a first isoprenoid sesquiterpene synthase polypeptide domain joined to a second different isoprenoid sesquiterpene synthase polypeptide domain claim 1 , interrupted by or including a ratio determinant domain claim 1 , whereby the chimeric isoprenoid sesquiterpene synthase polypeptide encoded by the DNA catalyzes the production of at least one isoprenoid sesquiterpene synthase reaction product that is not produced in the absence of the second ...

Подробнее
Номер записи: 23
25-04-2013 дата публикации

Method for producing patchoulol and 7-epi-alpha-selinene

Номер: US20130102038A1
Автор: Fabienne Deguerry, Michel Schalk
Принадлежит: FIRMENICH SA

A method of producing patchoulol and 7-epi-α-selinene by contacting at least one polypeptide with farnesyl phyrophosphate (fpp). The method may be carried out in vitro or in vivo to produce patchoulol and 7-epi-α-selinene, compounds which can be useful in the field of perfumery.

Подробнее
Номер записи: 24
02-05-2013 дата публикации

Transmembrane prostatic acid phosphatase

Номер: US20130108609A1
Автор: Pirkko Vihko
Принадлежит: CHEMPATH OY

The present invention relates to a novel transmembrane prostatic acid phosphatase (TM-PAP) protein or the C-terminal part thereof, nucleic acid molecules encoding said protein, vectors containing said nucleic acid molecules and host cells expressing said proteins. The present invention relates also to pharmaceutical compositions containing TM-PAP or the C-terminal part thereof and methods for using thereof in therapy and diagnostics. The present invention also relates to methods utilizing a transmembrane prostatic acid phosphatase knockout/knockdown non-human animal model and uses thereof.

Подробнее
Номер записи: 25
02-05-2013 дата публикации

PLANTS AND SEEDS WITH ALTERED STORAGE COMPOUND LEVELS, RELATED CONSTRUCTS AND METHODS INVOLVING GENES ENCODING PROTEINS WITH SIMILARITY TO BACTERIAL 2,4-DIHYDROXY-HEPT-2-ENE-1,7-DIOIC ACID CLASS II-LIKE ALDOLASE PROTEINS

Номер: US20130111633A1
Автор: MEYER KNUT, Stecca Kevin L.
Принадлежит: E.I. DU PONT DE NEMOOURS AND COMPANY

This invention is in the field of plant molecular biology. More specifically, this invention pertains to isolated nucleic acid fragments encoding proteins with similarity to bacterial 2,4-dihydroxy-hept-2-ene-1,7-dioic acid class II-like aldolase proteins in plants and seeds and the use of such fragments to modulate expression of a gene encoding proteins with similarity to bacterial 2,4-dihydroxy-hept-2-ene-1,7-dioic acid class II-like aldolase proteins in a transformed host cell. 136-. (canceled)37. A transgenic plant comprising a recombinant DNA construct comprising a polynucleotide operably linked to at least one regulatory element , wherein said polynucleotide encodes a polypeptide having an amino acid sequence of at least 85% sequence identity , based on the Clustal V method of alignment , when compared to SEQ ID NO: 29 , 31 , 33 , 35 , 49 , 107 , 108 , 109 , 110 , 111 , 112 , 113 , 114 , 115 , 116 , 117 , 118 , 119 , 120 , 121 , 122 , 123 , and 147 and wherein seed obtained from said transgenic plant has an altered i.e. increased or decreased oil , protein , starch and/or soluble carbohydrate content and/or altered seed weight when compared to a control plant not comprising said recombinant DNA construct.39. A method for producing a transgenic plant , the method comprising:(a) transforming a plant cell with a recombinant DNA construct comprising a polynucleotide operably linked to at least one regulatory sequence, wherein the polynucleotide encodes a polypeptide having an amino acid sequence of at least 85% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO: 29, 31, 33, 35, 49, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, and 123; and (b) regenerating a plant from the transformed plant cell and optionally (c) obtaining a progeny plant derived from transgenic plant, wherein said progeny plant comprises in its genome the recombinant DNA construct and seed obtained from said progeny plant ...

Подробнее
Номер записи: 26
02-05-2013 дата публикации

Methods and compositions for silencing genes using artificial micrornas

Номер: US20130111634A1
Автор: Brian Mcgonigle, Genhai Zhu, Itzhak Kurek
Принадлежит: E I Du Pont De Nemours And Co And Pioneer Hi-Bred International

Methods and compositions are provided that employ microRNA (miRNA) that, when expressed in a plant cell, is capable of reducing the level of mRNA of a target sequence (i.e. endogenous sequence) without reducing the level of mRNA of one or more closely related sequences. While miRNAs can be designed with specificity for a particular target sequence, the instant application demonstrates that a miRNA can specifically silence a target sequence without silencing a closely related sequence having high sequence identity to the target sequence. In certain embodiments, an endogenous target sequence can be suppressed with a recombinant miRNA expression construct without silencing a recombinant polynucleotide of interest having a sequence closely related to the target sequence. Such methods and compositions employ recombinant miRNA expression constructs which produce a 21-nt miRNA. Transgenic plant cells, plants and seeds incorporating miRNA expression constructs and recombinant polynucleotide constructs comprising polynucleotides of interest are also provided.

Подробнее
Номер записи: 27
09-05-2013 дата публикации

METHODS, COMPOSITIONS AND DEVICES UTILIZING STRUCTURALLY STABLE CYANURIC ACID HYDROLASE

Номер: US20130112608A1
Автор: Sadowsky Michael J., Seffernick Jennifer L., Wackett Lawrence P.
Принадлежит: Regents of the University of Minnesota

The present invention relates to stable cyanuric acid hydrolase enzymes, compositions, and devices for use in the treatment of a liquid, such as water. The present invention also relates to methods of using these enzymes, compositions and devices for the treatment of a liquid, such as water. 1. An isolated or purified structurally stable cyanuric acid hydrolase (CAH) enzyme.2. The CAH of having a Kvalue for cyanuric acid of 25-150 μM.3. The CAH of having a kvalue for cyanuric acid of 4.8-76 s.4. The CAH of claim 1 , wherein the enzyme is thermostable.5. The CAH of claim 1 , wherein the enzyme retains at least 30% enzymatic activity at a temperature above 25° C.6. The CAH of claim 1 , wherein the enzyme comprises amino acid sequence TEGNG(C/G/L)(V/M/A)ND(Y/F)(T/S)R (SEQ ID NO:15).7. The CAH of claim 1 , wherein the enzyme comprises amino acid sequence (M/F/L/I)(V/I/M)(M/F/W)SGG (D/E/G/P)G(V/I/L/G/A)(L/I/M/A)(S/T/A/C)PHX(T/I/L/S)(V/I/L)(F/I/V) (SEQ ID NO:17) claim 1 , wherein X is any amino acid.8. The CAH of claim 1 , wherein the enzyme comprises amino acid sequence TEGNGCVNDFTR (SEQ ID NO:16) and amino acid sequence FIMSGGEGVMTPHTVF (SEQ ID NO:18).9. The CAH of claim 1 , wherein the enzyme is 350-380 amino acids in length.10. The CAH of claim 1 , wherein the enzyme has a pI of about 5-6.11Moorella thermoacetica.. The CAH of claim 1 , wherein the enzyme is from12Moorella thermoacetica. The CAH of claim 11 , wherein the enzyme is from ATCC 39073.13. The CAH of claim 1 , wherein the enzyme has a specific activity of 12-18 μmmol/min/mg with cyanuric acid as substrate and less than 2 μmol/min/mg with barbituric acid as substrate.14. A composition for remediation of a liquid comprising the CAH of and polyethylene glycol (PEG) and/or KCl.15. The composition of claim 14 , wherein the PEG is PEG4000.16. The composition of claim 14 , wherein the PEG is present at a concentration of 5-50% by weight.17. The composition of claim 14 , wherein KCl is present at a concentration of ...

Подробнее
Номер записи: 28
09-05-2013 дата публикации

ISOLATED HIGH AFFINITY ENTITIES WITH T-CELL RECEPTOR LIKE SPECIFICITY TOWARDS NATIVE COMPLEXES OF MHC CLASS II AND GLUTAMIC ACID DECARBOXYLASE (GAD) AUTOANTIGENIC PEPTIDES

Номер: US20130115218A1
Автор: Dahan Rony, Reiter Yoram
Принадлежит: TECHNION RESEARCH & DEVELOPMENT FOUNDATION LTD

Provided are isolated complexes comprising a major histocompatibility complex (MHC) class II and a type I diabetes-associated GAD autoantigenic peptide, the isolated complex having a structural conformation which enables isolation of a high affinity entity which comprises an antigen binding domain capable of specifically binding to a native conformation of a complex composed of the MHC class II and the type I diabetes-associated GAD autoantigenic peptide; and isolated high affinity entities comprising an antigen binding domain capable of specifically binding the complex, wherein the isolated high affinity entity does not bind to the MHC class II in an absence of the diabetes-associated GAD autoantigenic peptide, wherein the isolated high affinity entity does not bind to the diabetes-associated GAD autoantigenic peptide in an absence of the MHC class II; and methods and kits using same for diagnostic and therapeutic purposes. 1. An isolated complex comprising a major histocompatibility complex (MHC) class II and a Glutamic acid decarboxylase (GAD) autoantigenic peptide , wherein said GAD autoantigenic peptide comprises a core amino acid sequence set forth by SEQ ID NO:14 (GAD556-565 , FFRMVISNPA) , wherein said GAD autoantigenic peptide is covalently attached to a beta chain of said MHC class II.2. The isolated complex of claim 1 , wherein said GAD autoantigenic peptide is covalently bound at a C terminus thereof to an N-terminus of an extracellular domain of a beta chain of said MHC class II.3. The isolated complex of claim 1 , wherein said GAD autoantigenic peptide is covalently embedded between amino acids 1-6 of an extracellular domain of a beta chain of said MHC class II.4. The isolated complex of claim 1 , wherein said GAD autoantigenic peptide is covalently attached to said beta chain between the third and forth amino acid of a mature polypeptide of said MHC class II beta chain.5. An isolated antibody comprising an antigen binding domain capable of ...

Подробнее
Номер записи: 29
09-05-2013 дата публикации

METHOD FOR THE GENERATION OF COMPACT TALE-NUCLEASES AND USES THEREOF

Номер: US20130117869A1
Автор: Bertonati Claudia, Duchateau Philippe, Epinat Jean-Charles, Juillerat Alexandre, Silva George H., Valton Julien
Принадлежит: CELLECTIS S.A.

The present invention relates to a method for the generation of compact Transcription Activator-Like Effector Nucleases (TALENs) that can efficiently target and process double-stranded DNA. More specifically, the present invention concerns a method for the creation of TALENs that consist of a single TALE DNA binding domain fused to at least one catalytic domain such that the active entity is composed of a single polypeptide chain for simple and efficient vectorization and does not require dimerization to target a specific single double-stranded DNA target sequence of interest and process DNA nearby said DNA target sequence. The present invention also relates to compact TALENs, vectors, compositions and kits used to implement the method. 1) A method for targeting and processing a double-stranded DNA , comprising:(a) selecting one DNA target sequence of interest on one strand of a double-stranded DNA; (i) one core TALE scaffold comprising Repeat Variable Dipeptide regions (RVDs) having DNA binding specificity onto said DNA target sequence of interest;', '(ii) at least one catalytic domain wherein said catalytic domain is capable of processing DNA a few base pairs away from said DNA target sequence of interest when fused to the C and/or N terminal of said core TALE scaffold from (i);', '(iii) optionally one peptidic linker to fuse said catalytic domain from (ii) to said core TALE scaffold from (i) when needed;, '(b) providing a unique compact TALEN monomer comprisingwherein said compact TALEN monomer is assembled to bind and process said double stranded DNA without requiring dirnerization;(c) contacting said double-stranded DNA with said unique monomer such that the double-stranded is processed a few base pairs away in 3′ and/or 5′ direction(s) from said one strand target sequence.2) A method according to claim 1 , wherein said catalytic domain has cleavage activity on said double-stranded DNA.3) A method according to claim 1 , wherein said catalytic domain is fused to ...

Подробнее
Номер записи: 30
16-05-2013 дата публикации

ENHANCED PHYTASE VARIANTS

Номер: US20130122567A1
Автор: Blesa Stephane, Chautard Helene, Delcourt Marc, Mesta Laurent, Winter Bruno
Принадлежит: BIOMETHODES

The present invention provides the field of enhancing proteins and in particular to that of proteins enhanced by molecular change. It provides a variant of a phytase that is termed enhanced in that is has better thermostability and/or activity than the original phytase. The invention also provides a nucleic acid coding for said variant, a cassette or an expression vector containing said variant, a host cell expressing said variant, a composition comprising said variant and uses thereof, principally in the preparation of food additives and animal feed. 1. An enhanced phytase variant whose sequence is SEQ ID NO: 1 or a functional derivative thereof , comprising at least one substitution on one of the amino acids from the group consisting of P3 , V4 , A5 , P8 , T9 , G10 , V16 , V17 , L19 , S20 , R21 , H22 , G23 , V24 , R25 , S26 , P27 , T28 , K29 , Q30 , T31 , Q32 , L33 , M34 , D36 , P39 , K41 , W45 , A49 , G50 , Y51 , L52 , T53 , G56 , A57 , V60 , Y67 , G75 , A78 , C81 , D92 , V93 , D94 , Q95 , R96 , T97 , R98 , L99 , T100 , G101 , A103 , V116 , V125 , D126 , F129 , H130 , P131 , V132 , D133 , D140 , T142 , Q143+H145 , A147 , L152 , P155 , L156 , E158 , E158+S160 , F167 , A177 , C182 , G189 , D193 , N196 , F197 , K201 , K206 , P207 , T209 , K210 , V211 , S212 , L213 , L217 , A218 , L219 , S220 , S221 , T222 , L223 , G224 , E225 , I226 , F227 , L228 , L229 , Q230 , N231 , Q233 , A234 , P236 , R242 , I250 , S251 , L252 , L253 , L255 , H256 , N257 , Q259 , F260 , D261 , M263 , A264 , Y268 , K273 , G274 , P276 , L277 , Q292 , G293 , P297 , P300 , Q301 , G308 , G309 , H310 , D311 , T312 , N313 , I314 , A315 , N316 , G322 , A323 , Q326 , P331 , D332 , N333 , T334 , P335 , P336 , G337 , G338 , G339 , V341 , E343 , D349 , Q352 , R353 , Y354 , I355 , A370 , E371 , K376 , P379 , A380 , G381 , D388 , E391 , S393 , G394 and P414 , the positions being indicated in SEQ ID NO: 1.2. The enhanced phytase variant whose sequence is SEQ ID NO: 1 or a functional derivative thereof ...

Подробнее
Номер записи: 31
23-05-2013 дата публикации

Methods and Compositions for Sequence Specific RNA Endonucleases

Номер: US20130129701A1
Автор: Choudhury Rajarshi, Wang Zefeng
Принадлежит:

The present invention provides sequence specific restriction enzymes for site-specific cleavage of RNA, as well as methods of their use. 1. A synthetic RNA endonuclease comprising the formula:{'br': None, 'A-B-C,'}wherein:A is an RNA binding domain,B is a linker peptide, andC is a cleavage domain.2. The RNA endonuclease of claim 1 , wherein the RNA binding domain comprises a Pumilio homology domain (PU-HUD).3. The RNA endonuclease of claim 2 , wherein the PU-HUD comprises a human Pumilio 1 (PUF) domain.4. The RNA endonuclease of claim 1 , wherein the linker peptide is from about three amino acids to about 20 amino acids in length.5. The RNA endonuclease of claim 1 , wherein the linker peptide comprises the amino acid sequence VDTGNGS.6. The RNA endonuclease of claim 1 , wherein the cleavage domain comprises the PilT N-terminus (PIN) domain of SMG6.7. The RNA endonuclease of claim 2 , wherein the PU-HUD is modified to bind an RNA sequence that is different than the RNA sequence bound by non-modified PU-HUD.8. The RNA endonuclease of claim 7 , wherein the RNA sequence is an 8 mer RNA sequence.9. The RNA endonuclease of claim 1 , wherein the RNA binding domain is at the amino terminus of the endonuclease and the cleavage domain as at the carboxy terminus of the endonuclease.10. The RNA endonuclease of claim 1 , comprising the amino acid sequence of ASRE(WT) claim 1 , ASRE(6-2/7-2) claim 1 , ASRE(6-2/7-2/1-1) claim 1 , ASRE(531) claim 1 , ASRE(87621) or ASRE(LacZ).11. A method of detecting a target RNA in a sample claim 1 , comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a) contacting the sample with the RNA endonuclease of under conditions whereby cleavage of RNA occurs if the target RNA is present in the sample and wherein the RNA binding domain of the RNA endonuclease is modified to bind the target RNA; and'}b) detecting a cleavage product of the target RNA, thereby detecting the target RNA in the sample.12. A method of cleaving a target mRNA in a ...

Подробнее
Номер записи: 32
23-05-2013 дата публикации

OBLIGATE HETERODIMER VARIANTS OF FOKI CLEAVAGE DOMAIN

Номер: US20130130350A1
Автор: CHANDRASEGARAN Srinivasan, KANDAVELOU Karthikeyan, RAMALINGAM Sivaprakash
Принадлежит:

Disclosed are methods of making and using engineered FokI cleavage domain variants. Also disclosed are methods, compositions and fusion proteins containing obligate heterodimers of engineered FokI cleavage domain variants and DNA binding domains, such as zinc finger protein (ZFP) domains and transcription activator-like effector (TALE) domains. 1. A polypeptide comprising an engineered FokI cleavage domain variant comprising a mutation in at least three or more wild-type amino acid residues , the engineered FokI cleavage domain variant forming an obligate heterodimer with a second engineered FokI cleavage domain variant with at least one different mutation in one or more wild-type amino acid residues.2. The polypeptide of claim 1 , wherein the amino acid residues at positions 483 claim 1 , 486 claim 1 , 487 claim 1 , 490 claim 1 , 499 or 538 are mutated.3. The polypeptide of claim 1 , wherein the engineered FokI cleavage domain variant comprises the polypeptide designated D483R:Q486E:I499L (SEQ ID NO: 1).4. The polypeptide of claim 1 , wherein the engineered FokI cleavage domain variant comprises the polypeptide designated R487D:E490K:I538K (SEQ ID NO: 3).5. The polypeptide of claim 1 , wherein the obligate heterodimer comprises a first monomer containing the polypeptide designated D483R:Q486E:I499L (SEQ ID NO: 1) claim 1 , and a second monomer containing the polypeptide designated R487D:E490K:I538K (SEQ ID NO: 3).6. The polypeptide of claim 1 , wherein the engineered FokI cleavage domain variant comprises the polypeptide designated D483R:Q486E:I499L:I538V (SEQ ID NO: 5).7. The polypeptide of claim 1 , wherein the engineered FokI cleavage domain variant comprises the polypeptide designated R487D:E490K:I499A:I538K (SEQ ID NO: 7).8. The polypeptide of claim 1 , wherein the obligate heterodimer comprises a first monomer containing the polypeptide designated D483R:Q486E:I499L:I538V (SEQ ID NO: 5) claim 1 , and a second monomer containing the polypeptide designated R487D ...

Подробнее
Номер записи: 33
23-05-2013 дата публикации

High Fidelity Restriction Endonucleases

Номер: US20130130935A1
Автор: Chan Siu-hong, Guan Shengxi, Quimby Aine, Sun Dapeng, Wei Hua, Xu Shuang-Yong, Zhang Penghua, Zhu Zhenyu
Принадлежит: NEW ENGLAND BIOLABS, INC.

Compositions and methods are provided for enzymes with altered properties that involve a systematic approach to mutagenesis and a screening assay that permits selection of the desired proteins. Embodiments of the method are particularly suited for modifying specific properties of restriction endonucleases such as star activity. The compositions includes restriction endonucleases with reduced star activity as defined by an overall fidelity index improvement factor. 1. A composition , comprising: a restriction endonuclease having at least one artificially introduced mutation and an overall fidelity index (FI) improvement factor of at least 2 , the restriction endonuclease being capable of cleaving a substrate with at least a similar cleavage activity to that of the restriction endonuclease absent the artificially introduced mutation , in a predetermined buffer , wherein the artificially introduced mutation is the product of at least one of a targeted mutation , saturation mutagenesis , or a mutation introduced through a PCR amplification procedure.2. A composition claim 1 , according to claim 1 , wherein at least one of the artificially introduced mutations is a replacement of a naturally occurring residue with an oppositely charged residue at a target site in the restriction endonuclease.3. A composition claim 1 , according to claim 1 , wherein at least one of the artificially introduced mutations is a replacement of a naturally occurring residue with a residue selected from a Phenylalanine and an Alanine at a target site in the restriction endonuclease.4. A composition claim 1 , according to claim 1 , wherein the restriction enzyme absent the at least one artificially introduced mutation is selected from the group consisting of: BamHI claim 1 , EcoRI claim 1 , ScaI claim 1 , SalI claim 1 , SphI claim 1 , PstI claim 1 , NcoI claim 1 , NheI claim 1 , SspI claim 1 , NotI claim 1 , SacI claim 1 , PvuII claim 1 , MfeI claim 1 , HindIII claim 1 , SbfI claim 1 , EagI claim ...

Подробнее
Номер записи: 34
06-06-2013 дата публикации

PRODUCTION OF MONOTERPENES

Номер: US20130143291A1
Автор: ALVIZO Oscar, McDaniel Robert, Meshulam-Simon Galit, Zhang Xiyun
Принадлежит: CODEXIS, INC.

The present invention relates to methods for producing monoterpenes, particularly tricyclene, by culturing microbial organisms that express a terpene synthase and optionally a prenyl transferase. 1. A method for producing tricyclene comprising:culturing a microbial organism expressing a heterologous terpene synthase under conditions in which the terpene synthase converts geranyl diphosphate to tricyclene, wherein the tricyclene is produced at a level of at least 0.5% of total monoterpene production by the organism.2. The method according to claim 1 , wherein the terpene synthase is a bornyl diphosphate synthase or a variant thereof.3. The method according to claim 1 , wherein the terpene synthase has at least 80% sequence identity to SEQ ID NO: 2 or 4.4. (canceled)5. The method according to claim 2 , wherein the terpene synthase comprises at least 90% sequence identity to SEQ ID NO: 2 claim 2 , and comprises an amino acid substitution at one or both of positions V399 and I404 of SEQ ID NO: 2.6. The method according to claim 5 , wherein the substitution at position V399 is V399I.7. The method according to claim 5 , wherein the terpene synthase further comprises an amino acid substitution at one or more of positions S4 claim 5 , E159 claim 5 , G338 claim 5 , S267 claim 5 , I291 claim 5 , I297 claim 5 , K285 claim 5 , T460 claim 5 , and F525 of SEQ ID NO: 2.8. The method according to claim 1 , wherein the terpene synthase is a camphene synthase or a variant thereof.9. The method according to claim 8 , wherein the terpene synthase has at least 80% sequence identity to SEQ ID NO: 6 or 8.10. (canceled)11. The method according to claim 8 , wherein the terpene synthase comprises at least 90% sequence identity with SEQ ID NO: 6 claim 8 , and comprises an amino acid substitution at one or more of positions N18 claim 8 , A283 claim 8 , I320 claim 8 , and T431 of SEQ ID NO: 6.12. The method according to claim 11 , wherein the terpene synthase further comprises an amino acid ...

Подробнее
Номер записи: 35
06-06-2013 дата публикации

MEGANUCLEASE VARIANTS CLEAVING A DNA TARGET SEQUENCE FROM THE DYSTROPHIN GENE AND USES THEREOF

Номер: US20130145487A1
Автор: Cedrone Frédéric
Принадлежит: CELLECTIS

The invention relates to meganuclease variants which cleave a DNA target sequence from the human dystrophin gene (DMD), to vectors encoding such variants, to a cell, an animal or a plant modified by such vectors and to the use of these meganuclease variants and products derived therefrom for genome therapy, ex vivo (gene cell therapy) and genome engineering including therapeutic applications and cell line engineering. The invention also relates to the use of meganuclease variants for inserting therapeutic transgenes other than DMD at the dystrophin gene locus, using this locus as a safe harbor locus. The invention also relates to the use of meganuclease variants for using the dystrophin gene locus as a landing pad to insert and express genes of interest. 1. An I-CreI variant , comprising at least two I-CreI monomers , wherein at least one of the two I-CreI monomers comprises at least two substitutions , one in each of two functional subdomains of a LAGLIDADG core domain situated from positions 26 to 40 and 44 to 77 of I-CreI , the variant being able to cleave a DNA target sequence from a dystrophin gene (DMD) , and wherein the I-CreI variant is obtained by a method comprising:(a) constructing a first series of I-CreI variants comprising a substitution of at least one position selected from the group consisting of 26, 28, 30, 32, 33, 38 and 40 of a first functional subdomain of the LAGLIDADG core domain situated from positions 26 to 40 of I-CreI,(b) constructing a second series of I-CreI variants comprising a substitution of at least one position selected from the group consisting of 44, 68, 70, 75 and 77 of a second functional subdomain of the LAGLIDADG core domain situated from positions 44 to 77 of I-CreI, (i) a nucleotide triplet in positions −10 to −8 of the I-CreI site has been replaced with a nucleotide triplet which is present in positions −10 to −8 of the DNA target sequence from DMD and', '(ii) a nucleotide triplet in positions +8 to +10 has been replaced ...

Подробнее
Номер записи: 36
13-06-2013 дата публикации

(r)-hydroxynitrile lyase from brassicaceae

Номер: US20130149752A1
Автор: Jennifer Andexer, Thorsten Eggert
Принадлежит: EVOCATAL GMBH

The invention concerns a polypeptide which can be isolated from the Brassicaceae family and which has at least the activity of a hydroxynitrile lyase (HNL). The hydroxynitrile lyase of the invention is the first HNL from the Brassicaceae family. The plants ( Arabidopsis ) from which this enzyme or its gene is isolated is also described as non-cyanogenic. All HNL-containing plants described so far are cyanogenic plants and so it has until now been assumed that only cyanogenic plants contain hydroxynitrile lyases. Surprisingly, it transpires that a polypeptide (AtHNL) of the invention is (R)-selective. The amino acid sequence gives a theoretical molecular weight of 29.2 kDa for the AtHNL subunit. The calculated molecular mass of the protein of approximately 30 kDa can be confirmed by SDS gel electrophoresis.

Подробнее
Номер записи: 37
13-06-2013 дата публикации

Heat-Stable Persephonella Carbonic Anhydrases and Their Use

Номер: US20130149771A1
Автор: Martin Simon Borchert
Принадлежит: Novozymes AS

The present invention relates to use of Persephonella carbonic anhydrase in CO 2 extraction, e.g., from flue gas, natural gas, biogas or ambient air. The Persephonella carbonic anhydrases are especially well suited for these purpose due to their extreme thermostability.

Подробнее
Номер записи: 38
13-06-2013 дата публикации

METHODS FOR ANALYZING LARIAT RNA

Номер: US20130150251A1
Автор: Menees Thomas Matthew
Принадлежит: The Curators of the University of Missouri

The present invention relates to compositions and methods useful for analyzing lariat RNA, which plays a role in the regulation of gene expression. A sample of RNA is specifically treated to remove linear mRNA and enrich for lariat RNA. The enriched lariat RNA sample may be analyzed further to identify introns, branch point sequences, alternative splicing patters, and gene transcription levels. The enriched lariat RNA sample may also be exploited as a detection or compound screening tool, as well as other uses. 1. An isolated debranching enzyme comprising an amino acid sequence having at least 35% sequence identity to a sequence selected from the group consisting of SEQ ID NO: 46-66.2. The isolated debranching enzyme of claim 1 , wherein the sequence identity is selected from the group consisting of about 40 claim 1 , 45 claim 1 , 50 claim 1 , 55 claim 1 , 60 claim 1 , 65 claim 1 , 70 claim 1 , 75 claim 1 , 80 claim 1 , 81 claim 1 , 82 claim 1 , 83 claim 1 , 84 claim 1 , 85 claim 1 , 86 claim 1 , 87 claim 1 , 88 claim 1 , 89 claim 1 , 90 claim 1 , 91 claim 1 , 92 claim 1 , 93 claim 1 , 94 claim 1 , 95 claim 1 , 96 claim 1 , 97 claim 1 , 98 claim 1 , 99% or more.3. The isolated debranching enzyme of claim 1 , wherein the amino acid sequence has at least 75% sequence identity to the metallophosphatase domain of a sequence selected from the group consisting of SEQ ID NO: 46-66.4. The isolated debranching enzyme of claim 3 , wherein the sequence identity is selected from the group consisting of about 40 claim 3 , 45 claim 3 , 50 claim 3 , 55 claim 3 , 60 claim 3 , 65 claim 3 , 70 claim 3 , 75 claim 3 , 80 claim 3 , 81 claim 3 , 82 claim 3 , 83 claim 3 , 84 claim 3 , 85 claim 3 , 86 claim 3 , 87 claim 3 , 88 claim 3 , 89 claim 3 , 90 claim 3 , 91 claim 3 , 92 claim 3 , 93 claim 3 , 94 claim 3 , 95 claim 3 , 96 claim 3 , 97 claim 3 , 98 claim 3 , 99% or more.5. A method of enriching an RNA population for lariat RNA comprising:a. providing an RNA population; and,b. ...

Подробнее
Номер записи: 39
20-06-2013 дата публикации

NOVEL ALDOLASE AND PRODUCTION PROCESS OF SUBSTITUTED ALPHA-KETO ACIDS

Номер: US20130157342A1
Автор: Kawahara Shigeru, Sugiyama Masakazu, Watanabe Kunihiko
Принадлежит: AJINOMOTO CO., INC.

4-(Indol-3-ylmethyl)-4-hydroxy-2-oxoglutarate, which is useful as an intermediate in the synthesis of monatin, may be synthesized from indole pyruvic acid and pyruvic acid (and/or oxaloacetic acid) by using a novel aldolase derived from the genus or 1. An isolated polynucleotide of following (a) or (b):(e) a polynucleotide having the nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15; or(f) a polynucleotide that hybridizes under stringent conditions with a polynucleotide having the nucleotide sequence complementary to the full-length nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15, and encodes a protein having aldolase activity, wherein said stringent conditions comprise a wash step in 0.1×SSC and 0.1% SDS at 65° C.2. An isolated polynucleotide of following (c) or (d):(g) a polynucleotide that encodes a protein comprising the amino acid sequence of SEQ ID NO: 16; or(h) a polynucleotide that encodes a protein having an amino acid sequence that contains a substitution, deletion, insertion, or addition of one to ten amino acid residues in the amino acid sequence of SEQ ID NO: 16, and has aldolase activity.3. A recombinant DNA produced by ligating into a vector DNA a polynucleotide selected from the group consisting of:(e) a polynucleotide having the nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15;(f) a polynucleotide that hybridizes under stringent conditions with a polynucleotide having the nucleotide sequence complementary to the full-length nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15, and encodes a protein having aldolase activity, wherein said stringent conditions comprise a wash step in 0.1×SSC and 0.1% SDS at 65° C.;(g) a polynucleotide that encodes a protein comprising the amino acid sequence of SEQ ID NO: 16; and(h) a ...

Подробнее
Номер записи: 40
20-06-2013 дата публикации

NUCLEOTIDE SEQUENCES AND CORRESPONDING POLYPEPTIDES CONFERRING MODULATED GROWTH RATE AND BIOMASS IN PLANTS GROWN IN SALINE CONDITIONS

Номер: US20130160163A1
Автор: FELDMANN Kenneth A., Sosa Julissa, Zhou Fasong
Принадлежит: CERES, INC.

The present invention relates to isolated nucleic acid molecules and their corresponding encoded polypeptides able confer the trait of improved plant size, vegetative growth, growth rate, seedling vigor and/or biomass in plants challenged with saline conditions. The present invention further relates to the use of these nucleic acid molecules and polypeptides in making transgenic plants, plant cells, plant materials or seeds of a plant having plant size, vegetative growth, growth rate, seedling vigor and/or biomass that are improved in saline conditions with respect to wild-type plants grown under similar conditions. 1. A method of modulating plant tolerance to salinity , said method comprising introducing into a plant cell at least one isolated nucleic acid comprising a nucleotide sequence selected from the group consisting of:(a) a nucleotide sequence that encodes an amino acid sequence and that is at least 85% identical SEQ ID NOs. 80, 99, 106, 123, 132, 146, 154 and 172, respectively;(b) a nucleotide sequence that is complementary to any one of the nucleotide sequences according to paragraph (a);(c) a nucleotide sequence according to any one of SEQ ID NOs. 79, 98, 105, 122, 131, 145, 153 and 171;(d) a nucleotide sequence that is an interfering RNA to the nucleotide sequence according to paragraph (a);(e) a nucleotide sequence able to form a hybridized nucleic acid duplex with the nucleic acid according to any one of paragraphs (a)-(d) at a temperature from about 5° C. to about 10° C. below a melting temperature of the hybridized nucleic acid duplex;(f) a nucleotide sequence encoding any one of the amino acid sequences identified as SEQ ID Nos. 80, 99, 106, 123, 132, 146, 154 and 172;{'figref': {'@idref': 'DRAWINGS', 'FIGS. 1-8'}, '(g) a nucleotide sequence encoding an amino acid sequence that fits an HMM based on the sequences aligned in any one of with an HMM bit score greater than 20; or'}{'figref': {'@idref': 'DRAWINGS', 'FIG. 9'}, '(h) a nucleotide sequence ...

Подробнее
Номер записи: 41
27-06-2013 дата публикации

METHOD FOR TARGETED GENOMIC EVENTS IN ALGAE

Номер: US20130164850A1
Автор: Sourdive David
Принадлежит:

The invention relates to endonucleases cleaving DNA target sequences from algae genomes, to appropriate vectors encoding such endonucleases, to cells or to algae modified by such vectors and to the use of these endonucleases and products derived therefrom for targeted genomic engineering in algae. 115-. (canceled)16. A method for targeted genomic engineering in an algal cell comprising introducing an endonuclease into the algal cell to induce a double-stranded cleavage at a site of interest in the genome of the algal cell.17. The method of claim 16 , comprising:providing an endonuclease capable of inducing a double-stranded cleavage at a site of interest in the genome of an algal cell;introducing the endonuclease into an algal cell; andisolating an algal cell having a modified targeted genomic site of interest.18. The method of claim 16 , wherein the endonuclease is introduced into the algal cell by electroporation or bombardment.19. The method of claim 17 , wherein the endonuclease is introduced into the algal cell by electroporation or bombardment.20. The method of claim 16 , wherein a targeted knock-out in algae is induced by the endonuclease at the site of interest in the genome.21. The method of claim 16 , wherein at least one transgene is inserted at the targeted genomic site of interest by introducing a template that is flanked by sequences sharing homology with the region surrounding the genomic DNA cleavage site of interest.22. The method of claim 21 , wherein the template comprises at least one transgene encoding a gene selected from the group consisting of quorum sensing claim 21 , secretion of hydrocarbons claim 21 , fatty acid composition claim 21 , lipids accumulation claim 21 , enhanced photosynthesis claim 21 , pigments production claim 21 , mercury volatilization claim 21 , frustule composition or organization claim 21 , and mitigation genes.23. The method of claim 21 , wherein the template comprises a nucleic acid encoding a selectable marker.24. ...

Подробнее
Номер записи: 42
04-07-2013 дата публикации

POLYPEPTIDE HAVING DITERPENE SYNTHASE ACTIVITY

Номер: US20130171701A1
Автор: Mitterbauer Rudolf, Specht Thomas
Принадлежит:

The present application, among others, relates to novel polypeptides having diterpene synthase activity, nucleic acid molecules encoding same, as well as to a gene cluster from which is thought to be involved in the biosynthetic pathway for producing pleuromutilin. 115.-. (canceled)16. An isolated polypeptide ,the polypeptide comprising an amino acid sequence which comprisesa sequence having at least 50% sequence identity to SEQ ID NO: 1,a sequence having at least 40% sequence identity to SEQ ID NO: 2, and i) a sequence having at least 15% sequence identity to SEQ ID NO: 7;', 'ii) a sequence having at least 25% sequence identity to SEQ ID NO: 4;', 'iii) a sequence having at least 45% sequence identity to SEQ ID NO: 5; and', 'iv) a sequence having at least 45% sequence identity to SEQ ID NO: 6,, 'at least one sequence selected from the group consisting of'}{'i': 'Clitopilus passeckerianus', 'wherein SEQ ID NOs: 1-2 and 4-7 are of origin and wherein said polypeptide is a diterpene synthase.'}17Clitopilus passeckerianus. The isolated polypeptide according to claim 16 , wherein said amino acid sequence further comprises a sequence having at least 50% sequence identity to SEQ ID NO: 3 claim 16 , wherein SEQ ID NO: 3 is of origin.18. The isolated polypeptide according to claim 16 , wherein the molecular weight of the polypeptide is between 90 kDa and 140 kDa.19. The isolated polypeptide according to claim 16 , wherein the polypeptide comprises an amino acid sequence which amino acid sequence comprises a sequence having at least 70% sequence identity to SEQ ID NO: 9.20. The isolated polypeptide according to claim 16 , wherein the polypeptide is capable of catalyzing the conversion of geranyl pyrophosphate into a pleuromutilin precursor.21. The isolated polypeptide according to claim 20 , wherein said pleuromutilin precursor is a compound according to formula (I).22. The isolated polypeptide according to claim 16 , wherein said polypeptide is derivable from a fungal host. ...

Подробнее
Номер записи: 43
04-07-2013 дата публикации

HIGHLY STABLE BETA-CLASS CARBONIC ANHYDRASES USEFUL IN CARBON CAPTURE SYSTEMS

Номер: US20130171718A1
Автор: ALVIZO Oscar, Benoit Michael R., Novick Scott J.
Принадлежит: CODEXIS, INC.

The present disclosure relates to β-class carbonic anhydrase polypeptides having improved properties including increased thermostability and/or stability in the presence of amine compounds, ammonia, or carbonate ion. The present disclosure also provides formulations and uses of the polypeptides for accelerating the absorption of carbon dioxide from a gas stream into a solution as well as for the release of the absorbed carbon dioxide for further treatment and/or sequestering. Also provided are polynucleotides encoding the carbonic anhydrase polypeptides and host cells capable of expressing them. 149-. (canceled)50. A recombinant carbonic anhydrase polypeptide comprising an amino acid sequence having at least 80% identity to SEQ ID NO: 2 and comprising an amino acid residue difference relative to SEQ ID NO: 2 at a position selected from X30 , X40 , X84 , X96 , X120 , X121 , X139 , and X147.51. The recombinant carbonic anhydrase polypeptide of in which the amino acid residue difference at a position selected from X30 claim 50 , X40 claim 50 , X84 claim 50 , X96 claim 50 , X120 claim 50 , X121 claim 50 , X139 claim 50 , and X147 is selected from X30A claim 50 , X30K claim 50 , X30L claim 50 , X30Q claim 50 , X30R claim 50 , X40L claim 50 , X40Q claim 50 , X40W claim 50 , X84K claim 50 , X84N claim 50 , X84Q claim 50 , X84R claim 50 , X84S claim 50 , X96A claim 50 , X96C claim 50 , X96E claim 50 , X96K claim 50 , X120R claim 50 , X121H claim 50 , X121K claim 50 , X121L claim 50 , X121Q claim 50 , X121T claim 50 , X121V claim 50 , X121W claim 50 , X139H claim 50 , X139K claim 50 , X139M claim 50 , X139Q claim 50 , X147E claim 50 , X147F claim 50 , X147G claim 50 , and X147T.52. The recombinant carbonic anhydrase polypeptide of in which the amino acid residue difference at a position selected from X30 claim 50 , X40 claim 50 , X84 claim 50 , X96 claim 50 , X120 claim 50 , X121 claim 50 , X139 claim 50 , and X147 is selected from X30R claim 50 , X40L claim 50 , X84Q claim ...

Подробнее
Номер записи: 44
18-07-2013 дата публикации

Fermentive production of four carbon alcohols

Номер: US20130183731A1
Автор: Andrew C. Eliot, Dennis Flint, Gail K. Donaldson, Lori Ann Maggio-Hall, Vasantha Nagarajan
Принадлежит: BUTAMAX ADVANCED BIOFUELS LLC

Methods for the fermentative production of four carbon alcohols is provided. Specifically, butanol, preferably isobutanol is produced by the fermentative growth of a recombinant bacterium expressing an isobutanol biosynthetic pathway.

Подробнее
Номер записи: 45
18-07-2013 дата публикации

MODIFIED ETHYLENEDIAMINE-N, N'-DISUCCINATE: ETHYLENEDIAMINE LYASE

Номер: US20130184448A1
Автор: Akiyama Takanori, MIZUNASHI Wataru, NAKAMURA Tetsuji
Принадлежит: MITSUBISHI RAYON CO., LTD.

The present invention provides a modified ethylenediamine-N,N′-disuccinate:ethylenediamine lyase. The present invention also provides a protein that comprises the amino acid sequence represented by SEQ ID NO: 1; or a protein that comprises an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 1 by deletion, substitution, or addition of one or more amino acid residues, and has an ethylenediamine-N,N′-disuccinate:ethylenediamine lyase activity. 153-. (canceled)54. An isolated gene encoding a protein comprising the amino acid sequence of SEQ ID NO:1 and having an ethylenediamine-N ,N′-disuccinate:ethylenediamine lyase activity.55. An isolated gene encoding a protein comprising the amino acid sequence of SEQ ID NO:1 having 1 to 10 substitutions , and wherein the protein has ethylenediamine-N ,N′-disuccinate:ethylenediamine lyase activity that is heat resistant.56. The isolated gene of claim 55 , wherein one substitution is the substitution of the lysine residue at position 120 with a glutamic acid residue.56. The isolated gene of claim 55 , wherein one substitution is the substitution of the isoleucine residue at position 166 with a serine residue.58. The isolated gene of claim 55 , wherein one substitution is the substitution of the isoleucine residue at position 166 with a threonine residue.59. The isolated gene of claim 55 , wherein one substitution is the substitution of the alanine residue at position 365 with a valine residue.60. An isolated gene encoding a protein comprising the amino acid sequence of SEQ ID NO:1 having 2 to 10 substitutions claim 55 , wherein the substitutions include the substitution of the lysine residue at position 120 with a glutamic acid residue claim 55 , and wherein the protein has ethylenediamine-N claim 55 ,N′-disuccinate:ethylenediamine lyase activity that is heat resistant.61. The isolated gene of claim 60 , wherein the isoleucine residue at position 166 is substituted with a serine residue.62. The ...

Подробнее
Номер записи: 46
25-07-2013 дата публикации

ANTIBODIES WITH T-CELL RECEPTOR LIKE SPECIFICITY TOWARDS NATIVE COMPLEXES OF MHC CLASS II AND DIABETES-ASSOCIATED AUTOANTIGENIC PEPTIDES

Номер: US20130189284A1
Автор: Dahan Rony, Reiter Yoram
Принадлежит: Technion Research & Development Foundation Ltd.

Provided are isolated complexes comprising a major histocompatibility complex (MHC) class II and a type I diabetes-associated autoantigenic peptide, the isolated complex having a structural conformation which enables isolation of a high affinity entity which comprises an antigen binding domain capable of specifically binding to a native conformation of a complex composed of the MHC class II and the type I diabetes-associated autoantigenic peptide; and isolated high affinity entities comprising an antigen binding domain capable of specifically binding the complex, wherein the isolated high affinity entity does not bind to the MHC class II in an absence of the diabetes-associated autoantigenic peptide, wherein the isolated high affinity entity does not bind to the diabetes-associated autoantigenic peptide in an absence of the MHC class II; and methods and kits using same for diagnostic and therapeutic purposes. 1. An isolated complex comprising a major histocompatibility complex (MHC) class II and a type I diabetes-associated autoantigenic peptide , the isolated complex having a structural conformation which enables isolation of a high affinity entity which comprises an antigen binding domain capable of specifically binding to a native conformation of a complex composed of said MHC class II and said type I diabetes-associated autoantigenic peptide , wherein said diabetes-associated autoantigenic peptide is covalently bound at a C terminus thereof to an N-terminus of an extracellular domain of a beta chain of said MHC class II.2. An isolated high affinity entity comprising an antigen binding domain capable of specifically binding a complex composed of a major histocompatibility complex (MHC) class II and a type I diabetes-associated autoantigenic peptide , wherein the isolated high affinity entity does not bind to said MHC class II in an absence of said diabetes-associated autoantigenic peptide , wherein the isolated high affinity entity does not bind to said diabetes- ...

Подробнее
Номер записи: 47
25-07-2013 дата публикации

MEGANUCLEASES VARIANTS CLEAVING A DNA TARGET SEQUENCE IN THE NANOG GENE AND USES THEREOF

Номер: US20130189759A1
Автор: Sourdive David
Принадлежит: CELLECTIS

Meganuclease variants cleaving DNA target sequences of the NANOG gene, vectors encoding such variants, and cells expressing them. Methods of using meganuclease variants recognizing NANOG gene sequences for modifying the NANOG gene sequence or for incorporating a gene of interest or therapeutic gene using the NANOG gene as a landing pad and a safe harbor locus. 1. A method for generating a secure iPS cell or a derivate thereof at various differentiation stages , the method comprising expressing at least one endonuclease in an iPS cell or a derivate thereof , wherein the at least one endonuclease induces a double-strand break in a NANOG gene to produce a cell lacking capacity for de-differentiation to a more pluripotent state.23-. (canceled)4. The method according to claim 1 , wherein said endonuclease is a meganuclease.5. A meganuclease variant that induces a double-strand break in a NANOG gene.6. The meganuclease of claim 5 , which recognizes the NANOG4 sequence (SEQ ID NO: 18).7. The meganuclease of claim 5 , which recognizes the NANOG4 sequence (SEQ ID NO: 18) and which comprises a variant I-CreI amino acid sequence selected from the group consisting of SEQ ID NO: 33 to 40.89-. (canceled)10. The meganuclease variant of claim 5 , which is a homodimer claim 5 , a heterodimer claim 5 , or a single chain.1114-. (canceled)15. The polynucleotide that encodes the meganuclease of or a fragment thereof having meganuclease activity.16. (canceled)17. A vector claim 15 , comprising the polynucleotide of .18. A host cell claim 17 , comprising the vector of .1928-. (canceled)29. A cell bank claim 17 , comprising cells in which NANOG is knocked-out by an endonuclease.30. A cell bank claim 17 , comprising cells in which NANOG is knocked-out by a meganuclease3134-. (canceled)35. A purified iPS cells culture claim 17 , wherein a NANOG gene of said iPS cells is not functional.36. A purified differentiated cell culture selected from the purified iPS cells culture according to .37. ...

Подробнее
Номер записи: 48
25-07-2013 дата публикации

Wheat plants having increased resistance to imidazolinone herbicides

Номер: US20130190170A1
Автор: Calvin Konzak
Принадлежит: Northwest Plant Breeding Co

The present invention is directed to wheat plants having increased resistance to an imidazolinone herbicide. More particularly, the present invention includes wheat plants containing one or more IMI nucleic acids such as a Gunner IMI 205, Gunner IMI 208 and Madsen IMI cultivar. The present invention also includes seeds produced by these wheat plants and methods of controlling weeds in the vicinity of these wheat plants.

Подробнее
Номер записи: 49
25-07-2013 дата публикации

High Fidelity Restriction Endonucleases

Номер: US20130190209A1
Автор: Chan Siu-hong, Guan Shengxi, Quimby Aine, Sun Dapeng, Wei Hua, Xu Shuang-Yong, Zhang Penghua, Zhu Zhenyu
Принадлежит: NEW ENGLAND BIOLABS, INC.

Compositions and methods are provided for enzymes with altered properties that involve a systematic approach to mutagenesis and a screening assay that permits selection of the desired proteins. Embodiments of the method are particularly suited for modifying specific properties of restriction endonucleases such as star activity. The compositions includes restriction endonucleases with reduced star activity as defined by an overall fidelity index improvement factor. 1. A composition , comprising: a restriction endonuclease having at least one artificially introduced mutation and an overall fidelity index (FI) improvement factor of at least 2 , the restriction endonuclease being capable of cleaving a substrate with at least a similar cleavage activity to that of the restriction endonuclease absent the artificially introduced mutation , in a predetermined buffer , wherein the artificially introduced mutation is the product of at least one of a targeted mutation , saturation mutagenesis , or a mutation introduced through a PCR amplification procedure.2. A composition claim 1 , according to claim 1 , wherein at least one of the artificially introduced mutations is a replacement of a naturally occurring residue with an oppositely charged residue at a target site in the restriction endonuclease.3. A composition claim 1 , according to claim 1 , wherein at least one of the artificially introduced mutations is a replacement of a naturally occurring residue with a residue selected from a Phenylalanine and an Alanine at a target site in the restriction endonuclease.4. A composition claim 1 , according to claim 1 , wherein the restriction enzyme absent the artificially introduced mutation is selected from the group consisting of: ScaI claim 1 , SphI claim 1 , PstI claim 1 , SspI claim 1 , PvuII claim 1 , MfeI claim 1 , SbfI claim 1 , EagI claim 1 , EcoRV claim 1 , AvrII claim 1 , BstXI claim 1 , PciI claim 1 , HpaI claim 1 , BsmBI claim 1 , BspQI claim 1 , SapI claim 1 , KpnI and ...

Подробнее
Номер записи: 50
01-08-2013 дата публикации

SYSTEMS AND METHODS FOR BIOTRANSFORMATION OF CARBON DIOXIDE INTO HIGHER CARBON COMPOUNDS

Номер: US20130196359A1
Автор: Baker David, Lidstrom Mary E., Louw Catherine, Siegel Justin, Smith Amanda Lee
Принадлежит: University of Washington through its Center for Commercialization

Systems, compounds and methods for the conversion of C1 carbon compounds to higher carbon compounds useful for the generation of commodity compounds. 1. A method for converting formaldehyde to dihydroxyacetone , comprising contacting formaldehyde with a thiamine diphosphate cofactor and a polypeptide comprising a motif capable of associating with the thiamine diphosphate cofactor ,wherein the motif comprises a histidine residue in a first position, an asparagine or glutamine residue in a second position, and a glutamic acid or aspartic acid residue in a third position;wherein upon association with the thiamine diphosphate cofactor, the cofactor pyrimidine N1 is hydrogen bonded to the acid side chain of the glutamic acid residue or the acid side chain of the aspartic acid residue of the motif; andwherein the histidine N1 and the amide side chain of the asparagine residue or the amide side chain of the glutamine residue are capable of making a water-mediated hydrogen bond with the formaldehyde substrate and/or dihydroxyacetone product.2. The method of claim 1 , wherein the motif is incorporated in a benzaldehyde lyase polypeptide scaffold.3. The method of claim 2 , wherein the benzaldehyde lyase polypeptide scaffold comprises an amino acid sequence with at least 40% identity to the sequence set forth in SEQ ID NO:2 or SEQ ID NO:3.4. The method of claim 1 , wherein the polypeptide is engineered.5. The method of claim 4 , wherein the motif comprises at least one amino acid substitution corresponding to the substitutions selected from the group consisting of A394G claim 4 , A480W claim 4 , G419N claim 4 , A28S claim 4 , and A28I claim 4 , with reference to the amino acid sequence set forth in SEQ ID NO:2.6. The method of claim 5 , wherein the motif comprises the amino acid substitutions corresponding to A394G and A480W claim 5 , with reference to the amino acid sequence set forth in SEQ ID NO:2.7. The method of claim 4 , wherein the motif comprises at least three amino ...

Подробнее
Номер записи: 51
01-08-2013 дата публикации

Method of Producing Dicer

Номер: US20130196383A1
Автор: Doudna Jennifer A., Ma Enbo
Принадлежит:

The present disclosure provides a method for producing a Dicer polypeptide in a prokaryotic host cell. The present disclosure further provides a purified Dicer complex. The present disclosure further provides kits for producing a Dicer polypeptide in a prokaryotic host cell. 1. A method of producing a Dicer polypeptide in a prokaryotic host cell , the method comprising:a) expressing a first Dicer polypeptide in a first prokaryotic host cell, wherein the first Dicer polypeptide comprises a DUF and a PAZ domain;b) expressing a second Dicer polypeptide in the first prokaryotic host cell or in a second prokaryotic host cell, wherein the second Dicer polypeptide comprises an RNAse IIIA domain and an RNase IIIb domain,wherein the first Dicer polypeptide and the second Dicer polypeptide spontaneously associate to form an enzymatically active Dicer complex.2. The method of claim 1 , wherein second Dicer polypeptide is expressed in the first prokaryotic host cell.3. The method of claim 1 , wherein the second Dicer polypeptide is expressed in a second prokaryotic host cell.4. The method of claim 1 , further comprising purifying the Dicer complex from the first and/or the second prokaryotic host cell.5. The method of claim 1 , wherein the first Dicer polypeptide comprises an amino acid sequence having at least about 85% amino acid sequence identity to amino acids 1-1008 claim 1 , amino acids 1-1068 claim 1 , amino acids 605-1008 claim 1 , amino acids 605-1068 claim 1 , amino acids 886-1008 claim 1 , or amino acids 886-1068 of the amino acid sequence set forth in claim 1 , and wherein the second Dicer polypeptide comprises an amino acid sequence having at least about 85% amino acid sequence identity to amino acids 1235 to about 1922 claim 1 , amino acids 1296 to 1922 claim 1 , amino acids 1235 to 1772 claim 1 , or amino acids 1296 to 1772 claim 1 , of the amino acid sequence set forth in .6. The method of claim 1 , wherein the first Dicer polypeptide has a length of from about ...

Подробнее
Номер записи: 52
08-08-2013 дата публикации

COMPOSITIONS AND METHODS FOR IMMUNIZATION AGAINST BACTERIA EXPRESSING A CARBAPENEMASE

Номер: US20130202616A1
Автор: Lin Lin, Spellberg Brad J.
Принадлежит: Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center

The present invention provides vaccine and pharmaceutical compositions for treating or preventing bacterial inventions. The vaccine compositions of the invention include a carbapenemase such as a serine carbapenemase, a metallo-β-lactamase or an immunogenic fragment thereof. The pharmaceutical compositions include an anti-carbapenemase antibody or fragment thereof. Also provided are methods for treating and preventing a bacterial infection using the vaccine and pharmaceutical compositions of the invention. The invention further provides antibody conjugates that include an antibody or fragment thereof conjugated to a siderophore or an analog thereof. 1. A vaccine composition comprising a serine carbapenemase or immunogenic fragment thereof and a pharmaceutically acceptable carrier.2K. pneumoniaSerratia marcescens. The vaccine composition of claim 1 , wherein said serine carbapenemase is selected from the group consisting of a carbapenemase (KPC) claim 1 , a enzyme (SME) claim 1 , a imipenem-hydrolyzing β-lactamase (IMI) claim 1 , a not metalloenzyme carbapenemase (NMC-A) claim 1 , an integron-borne cephalosporinase (IBC) and a Guiana extended spectrum (GES) enzyme.37-. (canceled)8. A method for treating or preventing a bacterial infection in a subject in need thereof comprising administering a therapeutically effective amount of a vaccine composition of to said subject.915-. (canceled)16. A pharmaceutical composition comprising an anti-serine carbapenemase antibody or a fragment thereof and a pharmaceutically acceptable carrier.1719-. (canceled)20. The pharmaceutical composition of claim 16 , wherein said anti-serine carbapenemase antibody is conjugated to a siderophore or an analog thereof.21. The pharmaceutical composition of claim 20 , wherein said siderophore is selected from the group consisting of enterobactin claim 20 , bacillibactin claim 20 , vibrobactin claim 20 , ferrichrome claim 20 , desferrioxamine claim 20 , fusarinine C claim 20 , ornibactin claim 20 ...

Подробнее
Номер записи: 53
08-08-2013 дата публикации

DETERMINATION OF IMMUNOGENIC PEPTIDES IN LYSOSOMAL ENZYMES AND INDUCTION OF ORAL TOLERANCE

Номер: US20130202633A1
Автор: Barrera Luis, Bellone Clifford, Knutsen Alan, Montaño-Suarez Adriana, Sosa-Molano Angela, Tomatsu Shunji
Принадлежит: Saint Louis University, a non-profit organization

Disclosed are methods and compositions for determining immunodominant peptides of target enzymes used in enzyme replacement therapy for lysosomal storage disorders. More specifically disclosed are immunodominant peptides for N-acetylgalactosamine-6-sulfatase (GALNS). Also disclosed are methods of inducing oral tolerance towards a target enzyme through oral administration of immunodominant peptides prior to commencing enzyme replacement therapy. More specifically disclosed is a method of inducing oral tolerance for GALNS, by orally administering specific immunodominant peptides for GALNS; in subjects suffering from mucopolysaccharidosis type IVA prior to commencing enzyme replacement therapy using GALNS. 1. An isolated peptide selected from the group consisting of SEQ ID NO:12 , SEQ ID NO:10 , SEQ ID NO:6 , fragments of SEQ ID NO:12 , fragments of SEQ ID NO:10 , and fragments of SEQ ID NO:6 , wherein said fragments consists of 6 amino acids or greater and said fragments are effective in inducing oral tolerance to N-acetyl galactosamine-6-sulfate sulfatase in a subject suffering from mucopolysaccharidosis type IVA.2. The isolated peptide of claim 1 , wherein said fragments consist of 12 amino acids or greater.3. The isolated peptide of claim 1 , wherein said fragments consist of 16 amino acids or greater.4. The isolated peptide of claim 1 , consisting of SEQ ID NO:12.5. The isolated peptide of claim 1 , consisting of SEQ ID NO:10.6. The isolated peptide of claim 1 , consisting of SEQ ID NO:6.7. The isolated peptide of claim 1 , further comprising fillers claim 1 , binders claim 1 , stabilizers claim 1 , preservatives claim 1 , buffers claim 1 , rapid release claim 1 , or sustained release components.8. A method of inducing oral tolerance to N-acetyl galactosamine-6-sulfate sulfatase in a subject suffering from mucopolysaccharidosis type IVA claim 1 , comprising claim 1 , administering by oral ingestion claim 1 , an effective amount claim 1 , for an effective period of ...

Подробнее
Номер записи: 54
08-08-2013 дата публикации

HEAT-STABLE CARBONIC ANHYDRASES AND THEIR USE

Номер: US20130203156A1
Автор: Borchert Martin, Saunders Paria
Принадлежит: NOVOZYMES A/S

The present invention relates to use of heat-stable carbonic anhydrase in COextraction, e.g., from flue gas, natural gas or biogas. Furthermore, the invention relates to isolated polypeptides having carbonic anhydrase activity at elevated temperatures and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides. 1Bacillus halodurans. A method of using a heat-stable carbonic anhydrase for extraction of carbon dioxide from a carbon dioxide-containing medium , comprising contacting a carbon dioxide-containing medium with a heat stable carbonic anhydrase that maintains activity at temperatures above 45° C. for at least 15 minutes , wherein the heat stable carbonic anhydrase is an alpha-carbonic anhydrase which is at least 85% identical to the alpha-carbonic anhydrase from (SEQ ID NO: 16) , wherein carbon dioxide is extracted from the carbon dioxide-containing medium.2Bacillus halodurans. The method in accordance with claim 1 , wherein the carbonic anhydrase is the alpha-carbonic anhydrase from (SEQ ID NO: 16).3. The method of claim 1 , where the carbonic anhydrase maintains activity at temperatures above 45° C. for at least two hours.4. The method of claim 1 , where the carbonic anhydrase is used in a bioreactor.5. The method of claim 4 , wherein the bioreactor comprises a contained liquid membrane.6. The method of claim 5 , wherein the membrane liquid of the contained liquid membrane contains a bicarbonate buffer with a pH of at least 9.0.7. The method of claim 1 , wherein the carbon dioxide-containing medium is a carbonic dioxide-containing gas.8. The method of claim 1 , wherein the carbon dioxide-containing medium is a multiphase mixture.9. The method of claim 7 , where the carbonic dioxide-containing gas is emitted from combustion or fermentation.10. The method of claim 7 , where the carbonic dioxide-containing gas is a flue gas.11. The method of claim 7 , where ...

Подробнее
Номер записи: 55
08-08-2013 дата публикации

Protoilludene synthase

Номер: US20130204034A1
Автор: Benedikt Engels, Marc Stadler, Stefan Jennewein, Torsten Grothe
Принадлежит: Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV

The present invention relates to an isolated, recombinant or synthetic polynucleotide encoding a polypeptide with protoilludene synthase activity and comprising a sequence selected from the group consisting of a) SEQ ID Nos. 1 or 14 of the attached sequence listing; b) a nucleic acid sequence complementary to SEQ ID Nos. 1 or 14; c) nucleic acid sequences which hybridize under stringent conditions to the nucleic acid sequences defined in a) and b) or their complementary strands, as well as to the polypeptide encoded by the isolated polynucleotide, as well as a method for the production of melleolides employing the polynucleotide or polypeptide of the invention.

Подробнее
Номер записи: 56
15-08-2013 дата публикации

INHIBITORS OF PDE4 BINDING TO HSP20

Номер: US20130209439A1
Автор: Baillie George S., Day Jon, Houslay Miles
Принадлежит: THE UNIVERSITY COURT OF THE UNIVERSITY OF GLASGOW

The present invention provides methods and materials for use in increasing HSP20 activation in a biological system, for example by increasing phosphorylation of Ser16 of HSP20. In one aspect, the invention provides a method for increasing HSP20 activation in a biological system, comprising contacting the system with an antagonist capable of inhibiting PDE4 binding to HSP20, the antagonist comprising or consisting essentially of a fragment of PDE4 or an analogue thereof. In a further aspect the invention provides a method of screening for an agent able to increase activation of HSP20. A preferred antagonist has a C-terminal lysine residue. 1. A method for increasing HSP20 activation in a biological system , comprising contacting the system with an antagonist capable of inhibiting PDE4 binding to HSP20.2. The method according to claim 1 , wherein the antagonist comprises an HSP20-binding moiety capable of inhibiting PDE4 binding to HSP20 claim 1 , wherein the HSP20-binding moiety is a fragment of PDE134 or analogue thereof.3. The method according to claim 2 , wherein the HSP20-binding moiety comprises an amino acid sequence represented by a formula selected from:X491-X492, X490-X491-X492, X489-X490-X491-X492, X488-X489-X490-X491-X492 or X487-X488-X489-X489-X490-X491-X492, wherein:X487 is F or is substituted with any amino acid other than Y, optionally an uncharged amino acid or a non-polar amino acid;X488 is Q or is substituted with any amino acid, optionally an uncharged amino acid or an uncharged polar amino acid;X489 is N or is substituted with any amino acid, optionally an uncharged amino acid or an uncharged polar amino acid;X490 is L or is substituted with any amino acid, optionally an uncharged amino acid or a non-polar amino acid;X491 is T or is substituted with any amino acid, optionally an uncharged amino acid or an uncharged polar amino acid;X492 is K.4. The method according to claim 3 , wherein:X487 is F or is substituted with an uncharged amino acid other ...

Подробнее
Номер записи: 57
15-08-2013 дата публикации

ASPARAGINASE FROM BASIDIOMYCETES

Номер: US20130209608A1
Автор: Berends Pieter, Berger Ralf Gunter, Eisele Nadine, Linke Diana, Rabe Swen
Принадлежит: NESTEC S.A.

An asparaginase enzyme derived from the fungi Basidiomycete, in particular the Basidiomycete is . A method for hydrolysing at least one of L-asparagine or L-glutamine. A method for reducing acrylamide formation in a substance comprising L-asparagine is also described. 1. An asparaginase enzyme obtainable from Basidiomycete.2Flammulina velutipes.. The asparaginase enzyme of claim 1 , wherein the Basidiomycete is Basidiomycete4. The asparaginase enzyme of in a form selected from the group consisting of a solid claim 1 , a liquid and an intermediate between a solid and a liquid.5. A combination of an asparaginase enzyme of obtainable from Basidiomycete and an exipient selected from the group consisting of lactose claim 1 , glycerol claim 1 , and albumin.6. A method for hydrolysing at least one of L-asparagine or L-glutamine comprising:treating a substance comprising at least one of L-asparagine or L-glutamine with an asparaginase enzyme obtainable from Basidiomycete.7. The method of claim 6 , wherein the substance comprising at least one of L-asparagine or L-glutamine is at least one of a human consumable product or an animal consumable product.8. The method of claim 6 , wherein the substance is selected from the group consisting of beverages claim 6 , cocoa beans claim 6 , cheese claim 6 , coffee beans claim 6 , confectionery claim 6 , desserts claim 6 , dough claim 6 , dressings claim 6 , French fries claim 6 , drinks claim 6 , meat products claim 6 , medical supplements claim 6 , nutritional supplements claim 6 , pet food claim 6 , potato chips claim 6 , sauces claim 6 , snacks and soups9. A method for reducing acrylamide formation in a food substance comprising L-asparagine comprising:applying to the food substance comprising L-asparagine an asparaginase enzyme obtainable from Basidiomycete; andheating the substance comprising L-asparagine.10. The method of claim 9 , wherein the substance is heated to at least 120° C.11. The method of claim 9 , wherein the source ...

Подробнее
Номер записи: 58
22-08-2013 дата публикации

FOOD ADDITIVE COMPRISING AN AMIDASE FOR DETOXIFYING OCHRATOXIN

Номер: US20130216654A1
Автор: Dalsgaard Søren, Nikolaev Igor, Poulsen Charlotte Horsmans, Wang Huaming, Yu Shukun
Принадлежит: DUPONT NUTRITION BIOSCIENCES APS

The present invention relates to amidase enzymes and a feed or food additive comprising the amidase enzyme capable of degrading ochratoxin. 1. An isolated amidase enzyme capable of degrading ochratoxin.2. The amidase according to claim 1 , wherein the ochratoxin is ochratoxin A.4. The amidase according to claim 1 , wherein the amidase has a Tim barrel structure including an active site comprising 6 histidine residues claim 1 , 1 lysine residue and 1 aspartic acid residue claim 1 , wherein the amino acid residues in the active site correspond to positions H111 claim 1 , H113 claim 1 , H191 claim 1 , K246 claim 1 , H287 claim 1 , H289 claim 1 , H307 and D378 of SEQ ID NO:1 when the tertiary structure of the amidase and SEQ ID NO:1 are compared.56-. (canceled)7. The amidase according to claim 1 , wherein the amidase enzyme comprises a polypeptide sequence having the sequence of any one of SEQ ID NO: 1 claim 1 , SEQ ID NO: 3 claim 1 , or SEQ ID NOs:5-11 claim 1 , 13 claim 1 , 14 or 15 claim 1 , or a sequence having at least 70% claim 1 , 75% claim 1 , 80% claim 1 , 90% claim 1 , 95% claim 1 , 98% claim 1 , 99% identity to any one of SEQ ID NO: 1 claim 1 , SEQ ID NO: 3 claim 1 , or SEQ ID NOs:5-11 claim 1 , 13 claim 1 , 14 or 15 claim 1 , or a polypeptide which differs from any one of SEQ ID NO: 1 claim 1 , SEQ ID NO: 3 claim 1 , or SEQ ID NOs:5-11 claim 1 , 13 claim 1 , 14 or 15 by one or several amino acid additions claim 1 , deletions and/or substitutions.8. The amidase according to claim 1 , wherein the amidase enzyme comprises a polypeptide sequence having the sequence of SEQ ID NO: 1 or SEQ ID NO: 3 or a sequence having at least 70% identity to SEQ ID NO: 1 or SEQ ID NO: 3 claim 1 , or a polypeptide which differs from SEQ ID NO: 1 or SEQ ID NO: 3 by one or several amino acid additions claim 1 , deletions and/or substitutions; or a polypeptide which is produced by expression of a polynucleotide comprising the sequence of SEQ ID NO: 2 or a sequence having at least 70 ...

Подробнее
Номер записи: 59
22-08-2013 дата публикации

Method of producing biofuel using brown algae

Номер: US20130217075A1
Автор: Byung Jo Yu, Hwa Young Cho, Jae Chan Park, Jae Hwa Lee, Sung Min Park
Принадлежит: SAMSUNG ELECTRONICS CO LTD

In a method of producing biofuel using brown algae, Bacterium antarctica is used as a hydrolysis catalyst for saccharification to obtain monosaccharides from the brown algae. The saccharification with the hydrolysis catalyst is effective in saccharification of the brown algae.

Подробнее
Номер записи: 60
29-08-2013 дата публикации

Phytase Variants

Номер: US20130224332A1
Автор: Andersen Carsten, De Maria Leonardo, Skov Lars Kobberoee, Soerensen Mikael Blom
Принадлежит: NOVOZYMES A/S

The present invention relates to a phytase which has at least 74% identity to a phytase derived from and comprises at least one alteration as compared to this phytase. These phytase variants have amended, preferably improved, properties, such as thermostability, temperature profile, pH profile, specific activity, performance in animal feed, reduced protease sensitiliby, and/or an amended glycosylation pattern. The invention also relates to DNA encoding these phytases, methods of their production, as well as the use thereof, e.g. in animal feed and animal feed additives. 129-. (canceled)30. A phytase variant , comprising an amino acid substitution corresponding to an amino acid substitution in SEQ ID NO: 2 at a position selected from the group consisting of 1 , 2 , 3 , 4 , 5 , 31 , 41 , 46 , 52 , 53 , 55 , 57 , 59 , 74 , 76 , 82 , 84 , 91 , 99 , 100 , 104 , 105 , 107 , 109 , 111 , 114 , 115 , 116 , 117 , 118 , 119 , 120 , 121 , 122 , 123 , 124 , 136 , 137 , 141 , 154 , 161 , 162 , 164 , 167 , 171 , 176 , 177 , 179 , 180 , 181 , 182 , 183 , 184 , 185 , 186 , 196 , 199 , 200 , 202 , 203 , 218 , 223 , 239 , 240 , 241 , 247 , 273 , 276 , 281 , 282 , 283 , 284 , 285 , 286 , 289 , 294 , 299 , 308 , 314 , 316 , 324 , 331 , 339 , 351 , 355 , 362 , 379 , 385 , 406 , 409 , 410 , and 411; provided that the phytase is not SEQ ID NO:3 , not SEQ ID NO:4 , and not SEQ ID NO:6 and wherein the variant has at least 80% identity to SEQ ID NO: 2 and has phytase activity.31. The phytase variant of claim 30 , which has at least 85% identity to SEQ ID NO: 2.32. The phytase variant of claim 30 , which has at least 90% identity to SEQ ID NO: 2.33. The phytase variant of claim 30 , which has at least 95% identity to SEQ ID NO: 2.35. A composition comprising the phytase of claim 30 , and(a) at least one fat soluble vitamin;(b) at least one water soluble vitamin; and/or(c) at least one trace mineral.36. The composition of claim 35 , further comprising at least one enzyme selected from the ...

Подробнее
Номер записи: 61
29-08-2013 дата публикации

Diterpene Synthases And Method For Producing Diterpenoids

Номер: US20130224809A1
Автор: Bohlmann Joerg, Zerbe Philipp
Принадлежит:

Provided herein are diterpene synthases (diTPS) and methods for producing diterpenoids. Also provided herein are nucleic acid sequences encoding diTPS, diTPS amino acid sequences, diTPS proteins, vectors, cells, transgenic organisms, uses, compositions, methods, processes, and kits thereof. 1. An isolated nucleic acid molecule , comprising a sequence of nucleotides encoding a cis-abienol synthase (CAS) polypeptide selected from among:a) the polypeptide whose sequence is set forth in one of SEQ ID NOS:7, 50 and 55;b) an active fragment of the polypeptide of a); andc) a polypeptide having a sequence of amino acids that has at least 85% sequence identity with a polypeptide of a) or b),wherein the encoded polypeptide or active fragment catalyzes the formation of cis-abienol from geranylgeranyl diphosphate (GGPP).2. The isolated nucleic acid molecule of claim 1 , wherein the sequence of nucleotides encodes a polypeptide that comprises the sequence of amino acids set forth in one of SEQ ID NOS:7 claim 1 , 50 and 55 or an active fragment thereof.3. The isolated nucleic acid molecule of claim 1 , wherein the sequence of nucleotides encodes a polypeptide that consists of the sequence of amino acids set forth in one of SEQ ID NOS:7 claim 1 , 50 and 55 or an active fragment thereof.4. The isolated nucleic acid molecule of claim 1 , wherein the active fragment is a pseudomature form.5. The isolated nucleic acid molecule of claim 4 , wherein the sequence of nucleotides encodes a cis-abienol synthase (CAS) polypeptide whose sequence is set forth in SEQ ID NO:50 or 55 claim 4 , or encodes a polypeptide whose sequence has at least 85% sequence identity with SEQ ID NO:50 or 55.6. The isolated nucleic acid molecule of claim 1 , comprising a sequence of nucleotides selected from among:a) the nucleic acid molecule whose sequence is set forth in one of SEQ ID NOS:8, 54 and 56; andb) a nucleic acid molecule whose sequence of nucleotides has at least 85% sequence identity to the sequence ...

Подробнее
Номер записи: 62
29-08-2013 дата публикации

Recombinant microorganisms and uses therefor

Номер: US20130224838A1
Автор: Fungmin Liew, Michael Koepke, Sean Simpson, Wendy Chen
Принадлежит: Individual

The invention provides, inter alia, methods for the production of acetone, isopropanol and/or precursors of acetone and/or isopropanol by microbial fermentation of substrates comprising CO, genetically modified microorganisms of use in such methods, nucleic acids suitable for preparation of genetically modified microorganisms, a novel alcohol dehydrogenase and nucleic acids encoding same.

Подробнее
Номер записи: 63
29-08-2013 дата публикации

USE OF ENDONUCLEASES FOR INSERTING TRANSGENES INTO SAFE HARBOR LOCI

Номер: US20130227715A1
Автор: Danos Olivier, Duclert Aymeric
Принадлежит: CELLECTIS

The present invention concerns the endonucleases capable of cleaving a target sequence located in a “safe harbor loci”, i.e. a loci allowing safe expression of a transgene. The present invention further concerns the use of such endonucleases for inserting transgenes into a cell, tissue or individual. 1. A variant endonuclease capable of cleaving a target sequence for use in inserting a transgene into a the genome of an individual , whereini. said genome comprises a locus comprising said target sequence; andii. said target sequence is located at a distance of at most 200 kb from a retroviral insertion site (RIS), wherein said RIS is neither associated with cancer nor with abnormal cell proliferation.2. The endonuclease according to claim 1 , wherein insertion of said transgene does not substantially modify expression of genes located in the vicinity of the target sequence.3. The endonuclease according to claim 1 , wherein said target sequence is located at a distance of at least 100 kb from the nearest genes.4. The endonuclease according to claim 1 , wherein said endonuclease is a homing endonuclease.5. The endonuclease according to claim 1 , wherein said endonuclease is capable of cleaving a target sequence located within a locus selected from the group consisting of the SH6 locus on human chromosome 21q21.1 claim 1 , the SH3 locus on human chromosome 6p25.1 claim 1 , the SH4 locus on human chromosome 7q31.2 claim 1 , the SH12 locus on human chromosome 13q34 claim 1 , the SH13 locus on human chromosome 3p12.2 claim 1 , the SH19 locus on human chromosome 22 claim 1 , the SH20 locus on human chromosome 12q21.2 claim 1 , the SH21 locus on human chromosome 3p24.1 claim 1 , the SH33 locus on human chromosome 6p12.2 claim 1 , the SH7 locus on human chromosome 2p16.1 claim 1 , the SH8 locus on human chromosome 5 claim 1 , the SH18 locus claim 1 , the SH31 locus claim 1 , the SH38 locus claim 1 , the SH39 locus claim 1 , the SH41 locus claim 1 , the SH42 locus claim 1 , the ...

Подробнее
Номер записи: 64
05-09-2013 дата публикации

ULVAN LYASE, METHOD FOR MANUFACTURING SAME, AND USES THEREOF

Номер: US20130230889A1
Автор: Helbert William, Lerat Yannick, Nyvall-Collen Pi, Sassi Jean-François
Принадлежит: UNIVERSITE PIERRE ET MARIE CURIE

The present invention notably relates to ulvan lyases, to nucleic acid sequences coding for these ulvan lyases, to vectors comprising these coding sequences, to a method of manufacturing these ulvan lyases, as well as to a method of degrading ulvans using these ulvan lyases and applicable applications to the degradation products of the ulvans. The ulvan lyases of the present invention, or ulvanolytic protein, are notably defined as proteins of 30 or 46 kD comprising the following four sequences in their peptide sequence: PNDPNLK, LLEVGNTGTFGSTGS, DLANPDNV and WNLPE. 2. The ulvan lyase according to claim 1 , said protein being of sequence SEQ ID No. 1.3. The ulvan lyase according to claim 2 , further comprising sequence SEQ ID No. 2 at its C-terminal end.4. The ulvan lyase according to claim 1 , further comprising a signal sequence at its N-terminal end.5. The ulvan lyase according to claim 4 , wherein the signal sequence is sequence SEQ ID No. 3.6. Nucleic acid of sequence SEQ ID No. 5.7. The nucleic acid according to claim 6 , further comprising sequence SEQ ID No. 6 at its 3′ end.8. The nucleic acid according to claim 6 , further comprising sequence SEQ ID No. 7 at its 5′ end.9. A vector comprising a nucleic acid claim 6 , the nucleic acid chosen sequence from SEQ ID No. 5 claim 6 , sequence SEQ ID No. 5 having sequence SEQ ID No. 6 at its 3′ end claim 6 , sequence ID No. 5 having sequence SEQ ID No. 7 at its 5′ end claim 6 , and sequence SEQ ID No. 5 having sequence SEQ ID No. 6 at its 3′ end and sequence SEQ ID No. 7 at its 5′ end.10. A host cell comprising a nucleic acid sequence or a vector claim 6 , the nucleic acid chosen from sequence SEQ ID No. 5 claim 6 , sequence SEQ ID No. 5 having sequence SEQ ID No. 6 at its 3′ end claim 6 , sequence ID No. 5 having sequence SEQ ID No. 7 at its 5′ end claim 6 , and sequence SEQ ID No. 5 having sequence SEQ ID No. 6 at its 3′ end and sequence SEQ ID No. 7 at its 5′ end claim 6 , and the vector comprising any of the ...

Подробнее
Номер записи: 65
12-09-2013 дата публикации

IDURONATE-2-SULFATASE AND USE THEREOF

Номер: US20130236442A1
Автор: LEE Sy, PARK Sung-Ick
Принадлежит: GREEN CROSS CORPORATION

Disclosed is a modified iduronate-2-sulfatase (IDS) gene constructed by inserting the nucleotide of SEQ ID NO: 2 into a wild-type IDS gene. In addition to being negatively charged, the improved IDS enzyme encoded by the modified gene exhibits a sufficient retention time in blood to target the bone, so that it is more effective for treating Hunter syndrome. 1. A gene comprising a coding sequence of a wild-type iduronate-2-sulfatase (IDS) gene , with an oligonucleotide encoding 5 to 7 negatively charged amino acids inserted into the coding sequence.2. The gene of claim 1 , wherein the oligonucleotide is inserted between an N-terminal leader sequence and a mature sequence of the IDS coding sequence.3. The gene of claim 1 , wherein the negatively charged amino acids are aspartic acid or glutamic acid.4. The gene of claim 1 , wherein the wild-type iduronate-2-sulfatase gene is represented by the nucleotide sequence as set forth in SEQ ID NO: 1 claim 1 , and the oligonucleotide is represented by the nucleotide sequence as set forth in SEQ ID NO: 2.5. The gene of claim 4 , wherein the oligonucleotide of SEQ ID NO: 2 is inserted between 75and 76bases of the polynucleotide of SEQ ID NO: 1.6. The gene of claim 2 , wherein a linker is further inserted between the oligonucleotide and the mature IDS coding sequence.7. The gene of claim 6 , wherein the linker has a nucleotide sequence of SEQ ID NO: 3.8. A polypeptide encoded by the gene of .9. An expression vector comprising the gene of .10. A host cell comprising the expression vector of .11. A pharmaceutical composition for treating or prevention of Hunter syndrome claim 8 , comprising the polypeptide of as an active ingredient.12. A method for treating or preventing Hunter syndrome claim 11 , comprising administering the composition of to a subject in need thereof. The present invention relates to an improved iduronate-2-sulfatase, and the use thereof.Hunter syndrome, or mucosaccharidosis type II, is a lysosomal storage ...

Подробнее
Номер записи: 66
12-09-2013 дата публикации

Manufacture of Active Highly Phosphorylated Human Lysosomal Sulfatase Enzymes and Uses Thereof

Номер: US20130236921A1
Автор: CHEN Zhi, Hague Charles, Pungor Erno
Принадлежит: Biomarin Pharmaceutical Inc.

This invention provides compositions of active highly phosphorylated lysosomal sulfatase enzymes, their pharmaceutical compositions, methods of producing and purifying such lysosomal sulfatase enzymes and compositions and their use in the diagnosis, prophylaxis, or treatment of diseases and conditions, including particularly lysosomal storage diseases that are caused by, or associated with, a deficiency in the lysosomal sulfatase enzyme. 138-. (canceled)39. A method for measuring the activity of a recombinant human lysosomal enzyme to degrade natural substrates , comprising:(a) culturing an isolated human cell deficient in the lysosomal enzyme under conditions in which natural substrates for the lysosomal enzyme accumulate;(b) contacting the cell with the lysosomal enzyme;(c) lysing the cell;(d) adding to the cell lysate an enzyme that (i) is specific for natural substrates and (ii) cleaves small oligosaccharides from the natural substrates;(e) labeling the small oligosaccharides with a detectable moiety;(f) optionally separating the labeled small oligosaccharides;(g) detecting the labeled small oligosaccharides; and(h) determining the activity of the lysosomal enzyme to degrade natural substrates by comparing (i) the amount of labeled small oligosaccharides from cells contacted with the lysosomal enzyme with (ii) the amount of labeled small oligosaccharides from cells not contacted with the lysosomal enzyme, wherein a reduction in (h)(i) as compared to (h)(ii) indicates the activity of the lysosomal enzyme to degrade natural substrates.40. The method of claim 39 , wherein the lysosomal enzyme is a lysosomal sulfatase enzyme selected from the group consisting of arylsulfatase B (ARSB) claim 39 , iduronate-2-sulfatase (IDS) claim 39 , sulfamidase/heparin-N-sulfatase (SGSH) claim 39 , N-acetylglucosamine-sulfatase (G6S) and N-acetylgalactosamine-ó-sulfatase (GALNS).41. The method of claim 40 , wherein the lysosomal sulfatase enzyme is GALNS.42. The method of claim 39 ...

Подробнее
Номер записи: 67
12-09-2013 дата публикации

MEGANUCLEASE VARIANTS CLEAVING A DNA TARGET SEQUENCE FROM THE MOUSE ROSA26 LOCUS AND USES THEREOF

Номер: US20130236946A1
Автор: Gouble Agnès
Принадлежит: CELLECTIS

An I-CreI variant, wherein one of the two I-CreI monomers has at least two substitutions, one in each of the two functional subdomains of the LAGLIDADG core domain situated respectively from positions 26 to 40 and 44 to 77 of I-CreI, said variant being able to cleave a DNA target sequence from the mouse ROSA26 locus. Use of said variant and derived products for the engineering of transgenic mice and recombinant mouse cell lines expressing an heterologous protein of interest. 143-. (canceled)44. A method of cleaving a DNA target sequence from a mouse ROSA26 locus comprising contacting said DNA target sequence with an I-CreI variant to thereby cleave said DNA target sequencewherein said I-CreI variant comprises a first monomer and a second monomer which are associated to form an active form,wherein said I-CreI variant comprises at least two substitutions in at least one of the monomers,wherein at least one substitution is of a residue in the range of positions 26 to 40 of I-CreI and at least one substitution is of a residue in the range of positions 44 to 77 of I-CreI andwherein said DNA target sequence is at least one sequence selected from the group consisting of SEQ ID NO: 5 to 14 and 16 to 30.45. The method of claim 44 , wherein said at least one substitution of a residue in the range of 26 to 40 of I-CreI is at least one substitution of a residue selected from the group consisting of positions 26 claim 44 , 28 claim 44 , 30 claim 44 , 32 claim 44 , 33 claim 44 , 38 and 40.46. The method of claim 44 , wherein said at least one substitution of a residue in the range of 44 to 77 of I-CreI is at least one substitution of a residue selected from the group consisting of positions 44 claim 44 , 68 claim 44 , 70 claim 44 , 75 and 77.47. The method of claim 44 , wherein said substitutions comprise replacing the wild-type amino acids with an amino acid selected from the group consisting of A claim 44 , D claim 44 , E claim 44 , G claim 44 , H claim 44 , K claim 44 , N ...

Подробнее
Номер записи: 68
12-09-2013 дата публикации

LEUCINE BETA ROLL DOMAINS AND USES THEREOF

Номер: US20130237612A1
Автор: Banta Scott
Принадлежит: The Trustees of Columbia University in the City of New York

In one aspect, the invention relates to a peptide that forms a calcium-dependent hydrogel using a rationally engineered beta roll peptide. In the absence of calcium, the peptide is intrinsically disordered. Upon addition of calcium, the peptide forms a corkscrew-like structure. In one embodiment, one face of the beta roll is mutated to comprise leucine residues. In some embodiments, a leucine zipper forming helical domain to the engineered beta roll forms hydrogels by physical cross-linking in calcium rich environments. 1. A beta roll comprising a scaffold from the RTX domain of adenylate cyclase , wherein leucine mutations are introduced on the beta roll domain.2. The beta roll of claim 1 , wherein the beta roll domain comprises the amino acid sequence XXXXXXXXX(SEQ ID NO: 10) claim 1 , wherein{'sub': '1', '(a) Xis an amino acid selected from the group consisting of glycine, valine and serine;'}{'sub': '2', '(b) Xis an amino acid selected from the group consisting of glycine, serine, aspartic acid and leucine;'}{'sub': '3', '(c) Xis an amino acid selected from the group consisting of alanine, glutamic acid, glutamine, tyrosine and glycine;'}{'sub': '4', '(d) Xis an amino acid selected from the group consisting of glycine and arginine;'}{'sub': '5', '(e) Xis an amino acid selected from the group consisting of aspartic acid, alanine, asparagine, serine, and histidine;'}{'sub': '6', '(f) Xis an amino acid selected from the group consisting of aspartic acid and asparagine;'}{'sub': '7', '(g) Xis an amino acid selected from the group consisting of valine, leucine, and threonine;'}{'sub': '8', '(h) Xis an amino acid selected from the group consisting of leucine, isoleucine, and tyrosine; and'}{'sub': '9', '(i) Xis an amino acid selected from the group consisting of isoleucine, leucine, serine, and arginine.'}3. The beta roll of claim 2 , wherein the beta roll domain further comprises X claim 2 , wherein Xis appended adjacent to X claim 2 , and wherein{'sub': '10', '(a) ...

Подробнее
Номер записи: 69
19-09-2013 дата публикации

Chimeric protein

Номер: US20130243765A1
Автор: Elliott A. Gruskin, Eric Theodore Cjojnicki, Gargi Roy
Принадлежит: Acorda Therapeutics Inc

A chimeric protein is disclosed for promoting repair and regeneration of neurons damaged by disease or physical injury wherein the chimeric protein is a combination of a first polypeptide possessing matrix modification activity and a second polypeptide possessing regenerating activity for neural cells.

Подробнее
Номер записи: 70
19-09-2013 дата публикации

Method for biocatalytic production of nitriles from oximes and oxime dehydratases usable therein

Номер: US20130244298A1
Автор: Andrea Piatesi, Kai-Uwe Baldenius, Wolfgang Siegel
Принадлежит: BASF SE

The present invention relates to novel methods for biocatalytic production of nitriles from oximes using oxime dehydratases and novel mutants with oxime dehydratase activity and use thereof in a process for biocatalytic production of nitriles, such as in particular for the production of citral nitrile, neral nitrile, geranial nitrile or citronellyl nitrile from citral oxime, neral oxime, geranial oxime or citronellal oxime; and oxime dehydratases usable therefor, nucleotide sequences therefor and expression constructs or microorganisms comprising these.

Подробнее
Номер записи: 71
19-09-2013 дата публикации

OVEREXPRESSION OF PHYTASE GENES IN YEAST SYSTEMS

Номер: US20130244303A1
Автор: Lei Xingen
Принадлежит: Cornell Research Foundation, Inc.

The present invention relates to a method of producing a heterologous protein or polypeptide having phytase activity in a yeast system. The invention also provides proteins having phytase activity which have increased thermostability. Yeast strains which produce a heterologous phytase and the vectors used to produce the phytase are also provided. 2E. coli. The method of wherein said polynucleotide encodes AppA phytase.3Pichia. The method of wherein said yeast is a species.4. The method of wherein said polynucleotide is carried on a nucleic acid vector. This application is a continuation application of U.S. patent application Ser. No. 12/941,445, filed Nov. 8, 2010, which is a continuation of U.S. patent application Ser. No. 11/962,446, filed Dec. 21, 2007, and issued as U.S. Pat. No. 7,829,318, on Nov. 9, 2010, which is a continuation application of U.S. patent application Ser. No. 11/372,851, filed Mar. 10, 2006, and issued as U.S. Pat. No. 7,312,063 on Dec. 25, 2007, which is a continuation application of U.S. patent application Ser. No. 10/094,693, filed Mar. 8, 2002, and issued as U.S. Pat. No. 7,026,150 on Apr. 11, 2006, which is a continuation-in-part of U.S. patent application Ser. No. 09/104,769, filed Jun. 25, 1998, and issued as U.S. Pat. No. 6,451,572 on Sep. 17, 2002, the contents of each of which are incorporated herein by reference in their entirety.The present invention relates to a method of producing phytase in yeast, yeast strains which express heterologous phytase, and the heterologous phytase produced by yeast.Phytases, a specific group of monoester phosphatases, are required to initiate the release of phosphate (“P”) from phytate (myo-inositol hexophosphate), the major storage form of P in cereal foods or feeds (Reddy, N. R. et al., “Phytates in Legumes and Cereals,” 28:1 (1982)). Because simple-stomached animals like swine and poultry as well as humans have little phytase activity in their gastrointestinal tracts, nearly all of the ingested ...

Подробнее
Номер записи: 72
19-09-2013 дата публикации

SOLVENT AND METHOD FOR CO2 CAPTURE FROM FLUE GAS

Номер: US20130244305A1
Автор: BABURAO BARATH, Bedell Stephen A., Ceperkovic Olivera, Fradette Sylvie, Versteeg Geert F., VITSE FREDERIC
Принадлежит:

The present disclosure describes the efficient use of a catalyst, an enzyme for example, to provide suitable real cyclic capacity to a solvent otherwise limited by its ability to absorb and maintain a high concentration of COcaptured from flue gas. This invention can apply to non-promoted as well as promoted solvents and to solvents with a broad range of enthalpy of reaction. 1. A solvent solution for the capture of COfrom a flue gas stream , the solvent solution including:an amine solvent; and{'sub': '2', 'a catalyst achieving increased COloadings in the amine solvent as compared to a non-catalyzed solvent at temperatures in the range of 80-140° F.'}2. The solvent solution of claim 1 , wherein the catalyst is a biocatalyst.3. The solvent solution of claim 1 , wherein the biocatalyst is carbonic anhydrase or an analog thereof.4. The solvent solution of claim 1 , wherein the amine solvent has a theoretical cyclic capacity greater than or equal to about 1 mole/liter.5. The solvent solution of claim 1 , wherein the amine solvent has an acid dissociation constant (pKa) greater than or equal to about 9 and less than or equal to about 10.5.6. The solvent solution of claim 1 , wherein the amine solvent is selected from the group including DMEA (dimethylethanolamine) claim 1 , DEEA (diethylethanolamine) claim 1 , and DMgly (dimethylglycine).7. A method of reducing energy demand of a system for capturing COfrom a flue gas stream using an amine solvent claim 1 , the method comprising:{'sub': 2', '2', '2', '2, 'claim-text': an amine solvent, and', {'sub': '2', 'a catalyst achieving increased COloadings in the amine solvent as compared to a non-catalyzed solvent at temperatures in the range of 80-140° F.; and'}], 'applying a COlean solvent solution to a COrich flue gas stream in an absorber column to provide a COrich solvent solution and a COlean flue gas stream, the solvent solution including{'sub': 2', '2, 'reducing a temperature of the COlean solvent solution provided to the ...

Подробнее
Номер записи: 73
19-09-2013 дата публикации

Als inhibitor herbicide tolerant beta vulgaris mutants

Номер: US20130247253A1
Автор: Andreas Loock, Bernd Holtschulte, Clemens Springmann, Guenter Donn, Juergen Benting, Nathalie Knittel-Ottleben, Rudorf Jansen, Ruediger Hain
Принадлежит: Bayer Intellectual Property GmbH

The present invention relates to an ALS inhibitor herbicide tolerant Beta vulgaris plant and parts thereof comprising a mutation of an endogenous acetolactate synthase (ALS) gene, wherein the ALS gene encodes an ALS polypeptide containing an amino acid different from tryptophan at a position 569 of the ALS polypeptide.

Подробнее
Номер записи: 74
26-09-2013 дата публикации

Purification of Cystathionine Beta-Synthase

Номер: US20130251700A1
Автор: CARRILLO Richard G., KRAUS Jan P., MAJTAN Thomas, NAVEH David
Принадлежит: THE REGENTS OF THE UNIVERSITY OF COLORADO

This invention provides chromatographic methods for the purification of a cystathionine β-Synthase (CBS) protein, particularly truncated variants thereof and compositions and pharmaceutical compositions prepared therefrom. 1. A method for purifying cystathionine β-Synthase (CBS) protein , wherein the CBS protein is a naturally occurring , chemically cleaved or genetically engineered truncated CBS protein , comprising the steps of:(a) providing a CBS-containing solution comprising one or a plurality of impurities; and(b) performing chromatographic separation of the CBS-containing solution using an ion exchange chromatography column and a metal affinity chromatography (IMAC) resin, wherein the impurities are removed thereby.2. The method of claim 1 , further comprising the step of performing chromatographic separation using a Hydrophobic Interaction Chromatography (HIC) column.3. The method of claim 1 , further comprising the step of performing chromatographic separation using a ceramic hydroxyapaptite resin.4. The method of claim 1 , wherein the ion exchange chromatography column is a weak anion exchanger.5. The method of claim 4 , wherein the weak anion exchanger is a DEAE-Sepharose FF column.6. The method of claim 1 , wherein the metal affinity chromatography (IMAC) resin is charged with a divalent metal cation.7. The method of claim 6 , wherein the divalent metal cation is nickel claim 6 , copper claim 6 , cobalt or zinc.8. The method of claim 7 , wherein the divalent metal ion is zinc.9. The method of claim 1 , further comprising eluting CBS from the metal affinity chromatography (IMAC) resin with an elution buffer comprising imidazole.10. The method of claim 1 , wherein the truncated CBS protein has an amino acid sequence identified by SEQ ID NO: 3.11. The method of claim 1 , wherein the CBS-containing solution is a clarified CBS solution.12. The method of claim 1 , wherein the CBS is produced in a recombinant cell.13. The method of claim 12 , wherein the ...

Подробнее
Номер записи: 75
26-09-2013 дата публикации

Three-dimensional structure of isoprene synthase and its use thereof for generating variants

Номер: US20130252303A1
Автор: Christopher L. Rife, Derek H. Wells, Jeffrey V. Miller, Richard R. Bott, Zachary Q. Beck
Принадлежит: DANISCO US INC, Goodyear Tire and Rubber Co

The present invention provides a three-dimensional structures of P. tremuloides isoprene synthase and P. alba isoprene synthase. The invention also provides methods of using the three-dimensional structure to design isoprene synthases with improved activity for increased isoprene production in microbial host cells. Biosynthetically produced isoprene of the present invention finds use in the manufacture of rubber and elastomers.

Подробнее
Номер записи: 76
26-09-2013 дата публикации

NOVEL ALDOLASE AND PRODUCTION PROCESS OF SUBSTITUTED ALPHA-KETO ACIDS

Номер: US20130252310A1
Автор: Kawahara Shigeru, Sugiyama Masakazu, Watanabe Kunihiko
Принадлежит: AJINOMOTO, CO., INC.

4-(Indol-3-ylmethyl)-4-hydroxy-2-oxoglutarate, which is useful as an intermediate in the synthesis of monatin, may be synthesized from indole pyruvic acid and pyruvic acid (and/or oxaloacetic acid) by using a novel aldolase derived from the genus , or 1. An isolated polynucleotide of following (a) or (b):(e) a polynucleotide having the nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15; or(f) a polynucleotide that hybridizes under stringent conditions with a polynucleotide having the nucleotide sequence complementary to the full-length nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15, and encodes a protein having aldolase activity, wherein said stringent conditions comprise a wash step in 0.1×SSC and 0.1% SDS at 65° C.2. An isolated polynucleotide of following (c) or (d):(g) a polynucleotide that encodes a protein comprising the amino acid sequence of SEQ ID NO: 16; or(h) a polynucleotide that encodes a protein having an amino acid sequence that contains a substitution, deletion, insertion, or addition of one to ten amino acid residues in the amino acid sequence of SEQ ID NO: 16, and has aldolase activity.3. A recombinant DNA produced by ligating into a vector DNA a polynucleotide selected from the group consisting of:(e) a polynucleotide having the nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15;(f) a polynucleotide that hybridizes under stringent conditions with a polynucleotide having the nucleotide sequence complementary to the full-length nucleotide sequence of SEQ ID NO: 15 or the nucleotide sequence of nucleotides 398 to 1141 of SEQ ID NO: 15, and encodes a protein having aldolase activity, wherein said stringent conditions comprise a wash step in 0.1×SSC and 0.1% SDS at 65° C.;(g) a polynucleotide that encodes a protein comprising the amino acid sequence of SEQ ID NO: 16; and(h) a ...

Подробнее
Номер записи: 77
03-10-2013 дата публикации

Means of controlling infection persistence of helicobacter pylori

Номер: US20130259888A1
Автор: Alma Fulurija, Barry J. Marshall, Douglas E. Berg, Mohammed Benghezal, Tobias Schoep
Принадлежит: ONDEK PTY LTD

The present invention relates to a means of controlling infection persistence of Helicobacter pylori ( H. pylori ). In particular, the present invention relates to an isolated, genetically modified Helicobacter pylori comprising a functional urease, wherein the contiguous amino acid sequence between amino acid 529 and amino acid 555 of SEQ ID NO:1 is altered to produce said modified Helicobacter pylori which is unable to establish or maintain a persistent infection.

Подробнее
Номер записи: 78
03-10-2013 дата публикации

ISOPRENE SYNTHASE VARIANTS FOR IMPROVED MICROBIAL PRODUCTION OF ISOPRENE

Номер: US20130260432A1
Автор: Bott Richard R., Cervin Marguerite A., JR. James T., Kellis, McAuliffe Joseph C., Miasnikov Andrei, PERES Caroline M., Rife Christopher Lee, WELLS Derek H., Weyler Walter, Whited Gregory M.
Принадлежит: DANISCO US INC.

The present invention provides methods and compositions comprising at least one isoprene synthase enzyme with improved catalytic activity and/or solubility. In particular, the present invention provides variant plant isoprene synthases for increased isoprene production in microbial host cells. Biosynthetically produced isoprene of the present invention finds use in the manufacture of rubber and elastomers. 156-. (canceled)57. An isolated host cell comprising a heterologous polynucleotide sequence encoding an isoprene synthase variant in operable combination with a promoter , wherein said isoprene synthase variant comprises one or more amino acid substitution(s) at one or more amino acid residues corresponding to a poplar isoprene synthase having the sequence of SEQ ID NO: 120 , wherein said substitution(s) are selected from the group consisting of V10M , F12S , T15A , E18G , V58I , V58F , L70Q , L70R , L70V , L70T , T71P , V79L , E89D , G94A , S119F , F120L , G127R , E175V , T212I , S257A , R262G , A266G , F280L , N297K , F305L , L319M , E323K , A328T , D342E , A359T , K366N , E368D , L374M , S396T , V4185 , K438N , H440R , T442A , I449V , A469S , K500R , K505Q , G507S , S509N , F511Y , and N532K; andwherein the variant is capable of more effectively converting dimethylallyl diphosphate (DMAPP) to isoprene, as compared to an isoprene synthase variant without a substitution.58. The host cell of wherein at least one amino acid substitution is a L70R substitution.59. The host cell of wherein at least one amino acid substitution is a G507S substitution.60. The host cell of wherein the variant comprises one of more amino acid substitutions selected from the group consisting of G127R/F511Y claim 57 , L70Q/G94A/R262G/F305L claim 57 , F12S/T15A/E18G/N297K claim 57 , S396T/T442I claim 57 , V10M/E323K claim 57 , F120L/A266G claim 57 , K438N/K500R claim 57 , V79L/S509N claim 57 , E175V/S257A/E368D/A469S claim 57 , T71P/L374M claim 57 , F280L/H440R claim 57 , E89D/H440R claim ...

Подробнее
Номер записи: 79
03-10-2013 дата публикации

ENZYMES THAT SYNTHESIZE ZINGIBERENE

Номер: US20130263329A1
Автор: Barry Cornelius, Gonzales-Vigil Eliana
Принадлежит: BOARD OF TRUSTEES OF MICHIGAN STATE UNIVERSITY

The invention relates to nucleic acids encoding a zingiberene synthase that enables host cells and plants to make zingiberene that is useful in fragrances and for repelling or killing insects. The invention also relates to isolated zingiberene synthases and to methods for making zingiberenes. 1. An isolated nucleic acid encoding a zingiberene synthase with at least 95% sequence identity to amino acid SEQ ID NO: 2 , 4 , 6 , 8 , 11 , 12 , 14 , 16 , 18 , or a combination thereof.2. The isolated nucleic acid of claim 1 , wherein the nucleic acid comprises a sequence with at least 86% sequence identity to any of nucleotide sequences SEQ ID NO:1 claim 1 , 3 claim 1 , 5 claim 1 , 7 claim 1 , 13 claim 1 , 15 claim 1 , 17 claim 1 , 19 claim 1 , or a combination thereof.3. An expression cassette comprising the nucleic acid of operably linked to a promoter functional in a host cell.4. A host cell comprising the nucleic acid of .5. The host cell of claim 4 , further comprising a promoter operably linked to the nucleic acid claim 4 , wherein the promoter is functional in the host cell.6. The host cell of claim 5 , wherein the host cell is a plant cell.7. The host cell of claim 5 , wherein the host cell is a microorganism.8. A plant tissue comprising the nucleic acid of .9. A plant tissue comprising the host cell of .10. A plant comprising the nucleic acid of .11. A plant comprising the plant tissue of .12. A method of making a zingiberene comprising:{'claim-ref': {'@idref': 'CLM-00004', 'claim 4'}, 'a) culturing the host cell of under conditions sufficient for expression of the zingiberene synthase; and'}b) providing the host cell with a substrate for the zingiberene synthase to make the sesquiterpene.13. The method of claim 12 , wherein the substrate is 2Z claim 12 ,6Z-farnesyl diphosphate.14. The method of claim 12 , wherein the host cell is a bacterial or yeast cell.15E. coli. The method of claim 14 , wherein the bacterial cell is an cell.16. An isolated zingiberene synthase ...

Подробнее
Номер записи: 80
10-10-2013 дата публикации

Coryneform bacterium transformant and process for producing phenol using the same

Номер: US20130267000A1
Автор: Hideaki Yukawa, Masayuki Inui
Принадлежит: Green Phenol Tech Res Association

Provided is a phenol-producing transformant constructed by transferring a gene which encodes an enzyme having tyrosine phenol-lyase activity into a coryneform bacterium as a host. Also provided is a process for producing phenol, which comprises a step of allowing the transformant to react in a reaction mixture containing tyrosine, a salt thereof, or an ester thereof under reducing conditions, and a step of collecting phenol from the reaction mixture.

Подробнее
Номер записи: 81
17-10-2013 дата публикации

1-deoxy-d-xylulose 5-phosphate synthase alleles responsible for enhanced terpene biosynthesis

Номер: US20130276166A1
Автор: Didier Merdinoglu, Eric Duchene, Phlippe Hugueney
Принадлежит: Genoplante Valor SAS

A method of enhancement of the 1-deoxy-D-xylulose 5-phosphate synthase (DXS) activity of plants or bacteria to increase terpenes production in cells, an enhanced DXS sequence likely to be obtained by this method, a method of enhancement of production of terpenes in a host cell containing the enhanced DXS enzyme, and transgenic bacterium or plants that express this polypeptide are described.

Подробнее
Номер записи: 82
24-10-2013 дата публикации

ENZYMES AND USES THEREOF

Номер: US20130280786A1
Автор: CLAVERIE JEAN-MICHEL, Joseph Pascale, Leonetti Jean-Paul, Roux Lucie
Принадлежит: DEINOVE

The present invention relates to novel enzymes and the uses thereof. The invention also relates to methods of producing such enzymes, coding nucleic acid molecules, recombinant cells and methods of transforming biomass from such materials. The invention is particularly suited to degrade biomass and/or to improve biomass degradation, and to produce bioenergy products or recombinant proteins. This invention also relates to various applications of the enzymes in the field of paper industry, textile industry as well as in the chemical and medical fields. 118-. (canceled)19. An isolated enzyme or polypeptide selected from:{'i': 'Deinococcus', 'a) an enzyme, wherein said enzyme is derived from a or a related bacterium and is involved in energetic metabolism; or'}b) a polypeptide comprising an amino acid sequence selected from SEQ ID NOs: 1-12, 27-41, 58, 60, 62, 64, 66, 68, 70 or 72 or a fragment thereof comprising at least 15 contiguous residues thereof.20. The isolated enzyme of claim 19 , which is active at a temperature of 30° C. or more.21. The isolated enzyme of claim 19 , wherein said enzyme catalyses biomass modification.22. The isolated enzyme of claim 21 , wherein said enzyme is selected from amylases claim 21 , glucosidases claim 21 , cellulases claim 21 , xylanases claim 21 , pectinases claim 21 , esterases claim 21 , acetyl xylan esterases claim 21 , or glucuronidases.23. The isolated enzyme of claim 22 , which comprises all or an active part of an amino acid sequence selected from SEQ ID NOs: 1-12 claim 22 , 58 claim 22 , 60 claim 22 , 62 claim 22 , 64 claim 22 , 66 claim 22 , 68 claim 22 , 70 or 72.24. The isolated enzyme of claim 21 , wherein said enzyme is involved in biofuel production by fermentation.25. The isolated enzyme of claim 24 , wherein said enzyme is selected from acetaldehyde dehydrogenases claim 24 , alcohol dehydrogenases or pyruvate dehydrogenases.26. The isolated enzyme of claim 24 , which comprises all or an active part of an amino acid ...

Подробнее
Номер записи: 83
07-11-2013 дата публикации

SYNTHETIC PATHWAY ENZYMES FOR THE PRODUCTION OF ARGYRINS

Номер: US20130295606A1
Автор: Garcia Ronald, Mueller Rolf, Wenzel Silke
Принадлежит:

The invention provides the amino acid sequences comprised in or constituting the synthetic pathway enzymes participating in the production of Argyrins, as well as the nucleic acid sequences encoding the synthetic pathway enzymes participating in the production of Argyrins, as well as genetically manipulated microorganisms containing nucleic acid sequences encoding the synthetic pathway enzymes for the production of Argyrins, e.g. for inserting one or more of these coding sequences, mutating in a targeted manner one or more of these nucleic acid sequences, in a wild type producer micro-organism or in a heterologous micro-organism, for the production of Argyrins. 1. An isolated nucleic acid molecule , encoding an amino acid sequence having enzymatic activity of synthetic pathway enzymes for the production of Argyrins , wherein the amino acid sequence comprises the amino acid sequences encoded by nucleotides 1608 to 4 of Seq.-ID No. 1 (orf1) , by nucleotides 3615 to 1687 of Seq.-ID No. 1 (orf2) , by nucleotides 5139 to 3661 of Seq.-ID No. 1 (orf3) , by nucleotides 7388 to 5274 of Seq.-ID No. 1 (orf4) , by nucleotides 7710 to 8048 of Seq.-ID No. 1 (orf5) , by nucleotides 8870 to 8043 of Seq.-ID No. 1 (orf6) , by nucleotides 9293 to 10282 of Seq.-ID No. 1 (orf7) , by nucleotides 11057 to 10320 of Seq.-ID No. 1 (orf8) , the amino acid sequences of Seq.-ID No. 7 (Arg1) , Seq.-ID No. 8 (Arg2) and Seq.-ID No. 9 (Arg3) , of Seq.-ID No. 10 (Arg4) and of Seq.-ID No. 11 (Arg5) , and the amino acid sequences encoded by nucleotides 45620 to 44706 of Seq.-ID No. 1 (orf9) , by nucleotides 46507 to 45617 of Seq.-ID No. 1 (orf10) , by nucleotides 47244 to 46504 of Seq.-ID No. 1 (orf11) , by nucleotides 47547 to 47975 of Seq.-ID No. 1 (orf12) , by nucleotides 48288 to 49268 of Seq.-ID No. 1 (orf13) , by nucleotides 49483 to 55209 of Seq.-ID No. 1 (orf14) and by nucleotides 55212 to 55565 of Seq.-ID No. 1 (orf15).2. The nucleic acid molecule according to claim 1 , having at least 90% ...

Подробнее
Номер записи: 84
07-11-2013 дата публикации

ISOPRENE SYNTHASE VARIANTS WITH IMPROVED SOLUBILITY FOR PRODUCTION OF ISOPRENE

Номер: US20130295632A1
Автор: RIFE Christopher L., WELLS Derek H.
Принадлежит:

The present invention provides methods and compositions of variant polypeptides having isoprene synthase activity with improved solubility. In particular, the present invention provides isoprene synthase variant for increased isoprene production in recombinant host cells.

Подробнее
Номер записи: 85
14-11-2013 дата публикации

THERAPEUTIC STRATEGIES TO TREAT CNS PATHOLOGY IN MUCOPOLYSACCHARIDOSES

Номер: US20130302308A1
Автор: Ballabio Andrea, Fraldi Alessandro
Принадлежит: FONDAZIONE TELETHON

The invention refers to nucleotide sequence encoding for a chimeric sulfatase, viral vectors expressing such sequences for gene therapy and pharmaceutical uses of the chimeric expressed protein. The invention is particularly applied in the therapy of mucopolysaccharidosis, preferably type IIIA. 1. A nucleotide sequence encoding for a chimeric sulfatase , said chimeric sulfatase consisting essentially in the N-terminal-C-terminal sequence order of:a) a signal peptide derived by either the human a-antitrypsin (hAAT) amino acid sequence or the human Iduronate-2-sulfatase (IDS) amino acid sequence;b) a human sulfatase derived amino acid sequence deprived of its signal peptide; andc) the ApoB LDLR-binding domain.2. The nucleotide sequence according to wherein the encoded signal peptide has a sequence belonging to SEQ ID No. 2 or SEQ ID No. 4.3. The nucleotide sequence according to wherein the human sulfatase is human sulfamidase.4. The nucleotide sequence according to wherein the encoded human sulfamidase derived amino acid sequence has essentially the sequence of SEQ ID No. 8.5. The nucleotide sequence according to wherein the encoded ApoB LDLR-binding domain has essentially the sequence of SEQ ID No. 10.6. The nucleotide sequence according to having essentially the sequence belonging to the following group:a) Assembly hAATsp-SGSH-3×flag cassette (SEQ ID No. 11),b) Assembly hIDSsp-SGSH-3×flag cassette (SEQ ID No. 13),c) Assembly hAATsp-SGSH-3×flag-ApoB cassette (SEQ ID No. 15),d) Assembly hIDSsp-SGSH-3×flag-ApoB cassette (SEQ ID No. 17),e) Assembly hAATsp-SGSH cassette (SEQ ID No. 19),f) Assembly hIDSsp-SGSH cassette (SEQ ID No. 21),g) Assembly hAATsp-SGSH-ApoB cassette (SEQ ID No. 23), andh) Assembly hIDSsp-SGSH-ApoB cassette (SEQ ID No. 25).7. A recombinant plasmid suitable for gene therapy of MPS comprising the nucleotide sequence according to under the control of a liver specific promoter.8. The recombinant plasmid according to wherein the liver specific promoter is ...

Подробнее
Номер записи: 86
14-11-2013 дата публикации

Polypeptides Having C4 Dicarboxylic Acid Transporter Activity and Polynucleotides Encoding Same

Номер: US20130302865A1
Автор: Amanda Fischer, Debbie Yaver
Принадлежит: Novozymes Inc

The present invention relates to isolated polypeptides having C4-dicarboxylic acid transporter activity and isolated polynucleotides encoding the polypeptides. 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, and methods of producing C4-dicarboxylic acids, such as malic acid.

Подробнее
Номер записи: 87
14-11-2013 дата публикации

Microorganisms and methods for the biosynthesis of adipate, hexamethylenediamine and 6-aminocaproic acid

Номер: US20130303723A1
Автор: Anthony P. Burgard, Mark J. Burk, Priti Pharkya, Robin E. Osterhout
Принадлежит: Genomatica Inc

The invention provides a non-naturally occurring microbial organism having a 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway. The microbial organism contains at least one exogenous nucleic acid encoding an enzyme in the respective 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway. The invention additionally provides a method for producing 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid. The method can include culturing a 6-aminocaproic acid, caprolactam or hexametheylenediamine producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding a 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway enzyme in a sufficient amount to produce the respective product, under conditions and for a sufficient period of time to produce 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid.

Подробнее
Номер записи: 88
28-11-2013 дата публикации

Method for Modifying a Property of a Protein

Номер: US20130316401A1
Автор: Gojobori Takashi, Hino Yumi, Hori Eiichi, Ikeo Kazuho, Kawarabayashi Yutaka, Kimura Eiichiro, Nakamura Yoji, Nishio Yosuke, Usuda Yoshihiro, Yamazaki Jun
Принадлежит: AJINOMOTO CO., INC.

A method for improving the thermostability of a protein includes introducing, into the protein, two or more amino acid substitutions selected from the group consisting of: (i) substitution of an arginine residue for a lysine residue, (ii) substitution of a threonine residue for a serine residue, and (iii) substitution of an alanine residue for a serine residue. 1. A method for improving thermostability of a protein , the method comprising introducing , into said protein , two or more amino acid substitutions selected from the group consisting of:(i) substitution of an arginine residue for a lysine residue,(ii) substitution of a threonine residue for a serine residue, and(iii) substitution of an alanine residue for a serine residue.2. The method according to claim 1 , wherein said protein is a coryneform bacterium protein.3Corynebacterium glutamicum. The method according to claim 2 , wherein said protein is a protein.4. A method for producing a protein having a modified thermostability claim 2 , the method comprising: (i) substitution of an arginine residue for a lysine residue,', '(ii) substitution of a threonine residue for a serine residue, and', '(iii) substitution of an alanine residue for a serine residue; and, '(a) introducing two or more amino acid substitutions into a gene encoding a protein, wherein the two or more amino acid substitutions are selected from the group consisting of(b) introducing said gene obtained in (a) into a suitable host for gene expression to express the protein having the modified thermostability.5. The method according to claim 4 , wherein said protein is a coryneform bacterium protein.6Corynebacterium glutamicum. The method according to claim 5 , wherein said protein is a protein.7. A method for producing a microorganism having a modified thermostability claim 5 , the method comprising introducing two or more amino acid substitutions into a chromosomal DNA of a microorganism to modify a thermostability of the microorganism claim 5 , ...

Подробнее
Номер записи: 89
05-12-2013 дата публикации

METHODS AND COMPOSITIONS FOR THE PRODUCTION OF EXTREMOPHILE ENZYMES FROM GREEN MICROALGAE AND CYANOBACTERIA

Номер: US20130323803A1
Автор: Grunden Amy Michele, Sederoff Heike Inge Ada
Принадлежит: North Carolina State University

The present invention relates to compositions and methods for stable transformation of green microalgae and for production of transgenic green microalgae and/or cyanobacteria that produce extremophile enzymes as co-products during the growth of the green microalgae and/or cyanobacteria for lipid biofuel production. Thus, the present invention provides nucleic acid constructs and methods of transformation useful in the production of stably transformed green microalgae and/or cyanobacteria expressing extremophile enzymes in combination with lipid production for biofuel. 123-. (canceled)24. A method for producing one or more extremophile enzymes , the method comprising:(a) culturing a green microalgae cell, wherein the green microalgae cell is stably transformed with a heterologous nucleotide sequence encoding one or more extremophile enzymes and expresses the one or more extremophile enzymes; and(b) collecting the one or more extremophile enzymes from the green microalgae cell culture of (a), thereby producing one or more extremophile enzymes.25. The method of claim 24 , wherein the green microalgae cell is stably transformed with the heterologous nucleotide sequence encoding one or more extremophile enzymes claim 24 , bypropelling the heterologous nucleotide sequence at a green microalgae cell embedded in a gel at a velocity sufficient to pierce the cell wall, cell membrane and chloroplast membrane and deposit the heterologous nucleotide sequence within a chloroplast of the green microalgae cell;wherein the heterologous nucleotide sequence is incorporated into the chloroplast genome of the green microalgae cell, thereby producing a stably transformed green microalgae cell, andfurther wherein the heterologous nucleotide sequence is carried by a microprojectile and the heterologous nucleotide sequence is propelled at the green microalgae cell by propelling the microprojectile at the green microalgae cell.26. The method of claim 24 , wherein the heterologous nucleotide ...

Подробнее
Номер записи: 90
12-12-2013 дата публикации

MUTANTS OF L-ASPARAGINASE

Номер: US20130330316A1
Автор: Bansal Saurabh, Kundu Bishwajit, Mishra Prashant
Принадлежит:

The present invention relates to a novel mutant of L-asparaginase enzyme characterized in having high thermostability, pH stability and no glutaminase activity useful for therapeutics and the process of preparing the same. The present invention specifically relates to mutant's MTCC 5580, MTCC 5581 and MTCC 5582 characterized in having higher stability, no glutaminase activity etc., to allow their usage in the form of improved protein therapeutics. A thermostable L-asparaginase from was cloned and expressed in host. The enzyme was engineered at its active site to create three different mutants based on structural and sequence analysis with a —derived enzyme homologue. The mutants MTCC 5580, MTCC 5581 and MTCC 5582 were tested for their stability, substrate affinity, optimum pH and temperature of activity and cytotoxicity. Based on the studies, all the three enzymes were found thermostable and with no glutaminase activity as compared to other available enzyme EcA II. MTCC 5579 and the above said three mutants showed the cytotoxicity on the leukemic cell lines. The present study showed that these enzymes are promising candidates for the treatment of leukemia. 1. A L-asparaginase comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 6 , SEQ ID NO: 8 , and SEQ ID NO: 10.2. The mutant of L-asparaginase of claim 1 , wherein the thermostability of said mutant is in the temperature range of 37-90° C.3. The mutant of L-asparaginase of claim 1 , wherein the pH stability of said mutant is in the range of 7.0-9.5.4. The mutant of L-asparaginase of claim 1 , wherein said mutant has no glutaminase activity.5. (canceled)6. A polynucleotide encoding the mutant of L-asparaginase of claim 1 , said polynucleotide comprising a sequence selected from the group consisting of SEQ ID NO: 7 claim 1 , SEQ ID NO: 9 claim 1 , and SEQ ID NO: 11.7. The mutant of L-asparaginase of claim 1 , which is encoded by a polynucleotide comprising a sequence amplified by primer ...

Подробнее
Номер записи: 91
12-12-2013 дата публикации

METHOD FOR PRODUCTION OF RECOMBINANT HUMAN IDURONATE 2-SULFATASE

Номер: US20130330802A1
Автор: Hatano Yuukichi, Kamei Shoichiro, Kawasaki Atsuko, Mihara Kazutoshi, Ootsuki Kouta, Sugimura Atsushi
Принадлежит: JCR PHARMACEUTICALS CO., LTD.

Disclosed is a method for production of recombinant human iduronate 2-sulfatase (rhI2S) in a large scale, with a high purity, and mannose 6-phosphate residues. The method comprises the steps of (a) culturing rhI2S-producing mammalian cells in a serum-free medium, (b) collecting culture supernatant, (c) subjecting the culture supernatant to cation-exchange column chromatography, (d) to dye affinity column chromatography, (e) to anion-exchange column chromatography, and (f) to a column chromatography employing as solid phase a material having affinity for phosphate group, and (g) to gel filtration column chromatography, in the order. 1. A method for production of recombinant human iduronate 2-sulfatase (rhI2S) comprising the steps of:(a) culturing rhI2S-producing mammalian cells in a serum-free medium to let them secrete rhI2S in the medium,(b) collecting culture supernatant by removing the cells from the culture that is obtained in step (a) above,(c) subjecting the culture supernatant collected in step (b) above to cation-exchange column chromatography to collect rhI2S-active fractions,(d) subjecting the fractions collected in step (c) above to dye affinity column chromatography to collect rhI2S-active fractions,(e) subjecting the fractions collected in step (d) above to anion-exchange column chromatography to collect rhI2S-active fractions,(f) subjecting the fractions collected in step (e) above to a column chromatography employing as solid phase a material having affinity for phosphate group to collect rhI2S-active fractions, and(g) subjecting the fractions collected in step (f) above to gel filtration column chromatography to collect rhI2S-active fractions, in the order.2. The method according to claim 1 , wherein the cation exchanger employed in the cation-exchange column chromatography is a weak cation exchanger.3. The method according to claim 2 , wherein the weak cation exchanger has a selectivity based on both hydrophobic interaction and hydrogen bond ...

Подробнее
Номер записи: 92
12-12-2013 дата публикации

Recombinant microorganisms and uses therefor

Номер: US20130330809A1
Автор: Alexander Paul MUELLER, Michael Koepke, Shilpa NAGARAJU
Принадлежит: Lanzatech New Zealand Ltd

Carboxydotrophic acetogenic microorganisms do not produce MEK and/or 2-butanol. They lack the biosynthesis pathways to make these products. In addition, they produce the intermediate (R,R)-2,3-butanediol whereas the production of MEK and 2-butanol requires production of the intermediate (R,S)-2,3-butanediol. Nonetheless, the production of MEK and/or 2-butanol can be accomplished using recombinant microorganisms adapted to express or overexpress key enzymes in the MEK and/or 2-butanol biosynthesis pathways. Such microorganisms, such as the carboxydotrophic acetogen Clostridium autoethanogenum , can ferment substrates comprising CO. The overall scheme involves the production of 2-butanol from (R,S)-2,3-butanediol and the conversion of (R)-acetoin to (S)-2,3-butanediol. These steps are involved in the production of both MEK and 2-butanol. Such fermentation methods offer a means of using carbon monoxide from industrial processes which would otherwise be released into the atmosphere and pollute the environment.

Подробнее
Номер записи: 93
19-12-2013 дата публикации

METHOD FOR INCREASING THE EFFICIENCY OF DOUBLE-STRAND BREAK-INDUCED MUTAGENESIS

Номер: US20130337454A1
Автор: Duchateau Philippe, Epinat Jean-Charles, Juillerat Alexandre, Silva George H.
Принадлежит:

The present invention relates to a method for increasing double-strand break-induced mutagenesis at a genomic locus of interest in a cell, thereby giving new tools for genome engineering, including therapeutic applications and cell line engineering. More specifically, the present invention concerns a method for increasing double-strand break-induced mutagenesis at a genomic locus of interest, leading to a loss of genetic information and preventing any scarless re-ligation of said genomic locus of interest by NHEJ. The present invention also relates to engineered endonucleases, chimeric or not, vectors, compositions and kits used to implement this method. 151-. (canceled)52. A method for increasing double-strand break induced mutagenesis at a genomic locus of interest in a cell comprising the steps of:(i) identifying at said genomic locus of interest at least one DNA target sequence cleavable by one rare-cutting endonuclease;(ii) engineering said at least one rare-cutting endonuclease in order to generate a loss of genetic information around said DNA target sequence within the genomic locus of interest; and(iii) contacting said DNA target sequence with said at least one rare-cutting endonuclease to generate said loss of genetic information around said DNA target sequence within the genomic locus of interest;thereby obtaining a cell in which double-strand break induced mutagenesis at said genomic locus of interest is increased.53. The method according to claim 52 , wherein said engineered rare-cutting endonuclease is a chimeric rare-cutting endonuclease comprising a catalytic domain selected from table 2 (SEQ ID NO: 38-57) and table 3 (SEQ ID NO: 96-152) claim 52 , a functional mutant claim 52 , a variant or a derivative thereof.54. The method according to claim 53 , wherein said chimeric rare-cutting endonuclease comprises a catalytic domain selected from the group of Trex (SEQ ID NO: 145-149) and Tdt (SEQ ID NO: 201) claim 53 , a functional mutant claim 53 , a ...

Подробнее
Номер записи: 94
19-12-2013 дата публикации

CLASS II HUMAN HISTONE DEACETYLASES, AND USES RELATED THERETO

Номер: US20130338024A1
Автор: Grozinger Christina M., Hassig Christian A., Schreiber Stuart L.
Принадлежит: President and Fellows of Harvard College

The invention provides histone deacetylase class II nucleic acids and polypeptides, methods and reagents for their use, and related compounds including small molecule libraries containing class II histone deacetylase inhibitors. 1. An isolated or recombinant polypeptide , wherein the polypeptide comprises an HDx polypeptide sequence at least 88 percent identical to SEQ ID NO: 4 , or fragment thereof.2. The polypeptide of claim 1 , wherein the polypeptide is encoded by a nucleic acid having a coding sequence claim 1 , or portion thereof claim 1 , which hybridizes under stringent conditions to the nucleic acid sequence designated in SEQ ID NO: 3.3. The polypeptide of claim 1 , wherein the polypeptide comprises an HDx polypeptide sequence at least 95 percent identical to SEQ ID NO: 4 claim 1 , or fragment thereof.4. The polypeptide of claim 1 , wherein the polypeptide comprises an HDx polypeptide sequence at least 98 percent identical to SEQ ID NO: 4 claim 1 , or fragment thereof.5. The polypeptide of claim 1 , wherein the polypeptide comprises an HDx polypeptide sequence at least 99 percent identical to SEQ ID NO: 4 claim 1 , or fragment thereof.6. The polypeptide of claim 1 , wherein the polypeptide comprises an HDx polypeptide sequence identical to SEQ ID NO: 4.7. The polypeptide of claim 1 , wherein the polypeptide is of mammalian origin.8. The polypeptide of claim 1 , wherein polypeptide is of human origin.9. The polypeptide of claim 1 , wherein the polypeptide has deacetylase activity.10. The polypeptide of claim 1 , wherein the polypeptide binds to a histone claim 1 , a 14-3-3 protein claim 1 , a MEF2 transcription factor claim 1 , or a retinoblastoma associated protein such as RbAp48.11. The polypeptide of claim 1 , wherein the polypeptide is a fusion protein.12. The polypeptide of claim 11 , wherein the polypeptide comprises the HDx polypeptide claim 11 , or fragment thereof claim 11 , fused to glutathione-S-transferase.13. The polypeptide of claim 11 , ...

Подробнее
Номер записи: 95
26-12-2013 дата публикации

Recombinant Colwellia Psychrerythraea Alkaline Phosphatase and Uses Thereof

Номер: US20130344493A1
Автор: Kubu Christopher J., Muller-Greven Jeannine C., Post Marc A.
Принадлежит: Affymetrix, Inc.

A heat labile alkaline phosphatase enzyme and methods of using the same and kits including the same are disclosed. Specifically, a nucleotide sequence of, peptide sequence of, methods of using, and kits comprising, a heat labile alkaline phosphatase isolated from are provided. Methods of over-expression and purification of the recombinant alkaline phosphatase and mutants thereof are also disclosed. Methods of over-expressing and purifying commercially useful quantities of active recombinant heat labile alkaline phosphatase fusion enzymes from wherein the fusion enzymes comprise one or more heterologous leader sequences are disclosed. The disclosed heat labile alkaline phosphatase has properties similar to shrimp alkaline phosphatase and can be substituted for shrimp alkaline phosphatase in assays involving the same. 1. An isolated nucleic acid comprising:a) a recombinant sequence which is at least 95% identical to SEQ ID NO:3;b) the sequence which is fully complementary to SEQ ID NO:3;c) the sequence which is fully complementary to the sequence which is at least 95% identical to SEQ ID NO:3; ord) the sequence which hybridizes to SEQ ID NO:3 under stringent conditions.23-. (canceled)4. An isolated recombinant protein comprising the sequence according to SEQ ID NO:15 , or the sequence at least 95% identical to SEQ ID NO:15.56-. (canceled)7. A method of dephosphorylating a compound comprising a phosphate group , which comprises incubating a compound comprising a phosphate group with an effective amount of an isolated enzyme encoded by:a) the sequence according to SEQ ID NO:3;b) a sequence which is at least 95% identical to SEQ ID NO:3;c) the sequence which is fully complementary to SEQ ID NO:3;d) the sequence which is fully complementary to the sequence which is at least 95% identical to SEQ ID NO:3; ore) the sequence which hybridizes to SEQ ID NO:3 under stringent conditions.8. The method according to claim 7 , wherein said compound is a nucleic acid claim 7 , DNA or ...

Подробнее
Номер записи: 96
02-01-2014 дата публикации

PURIFICATION OF IDURONATE-2-SULFATASE

Номер: US20140004096A1
Автор: Nichols Dave
Принадлежит:

The present invention provides, among other things, improved methods for purifying I2S protein produced recombinantly for enzyme replacement therapy. The present invention is, in part, based on the surprising discovery that recombinant I2S protein can be purified from unprocessed biological materials, such as, I2S-containing cell culture medium, using a process involving as few as four chromatography columns. 1. A composition comprising purified recombinant iduronate-2-sulfatase (I2S) having an amino acid sequence at least 70% identical to SEQ ID NO:1 ,wherein the purified recombinant I2S comprises at least about 70% conversion of the cysteine residue corresponding to Cys59 of SEQ ID NO:1 to Cα-formylglycine (FGly) andfurther wherein the purified recombinant I2S contains less than 150 ng/mg Host Cell Protein (HCP).23.-. (canceled)5. The composition of claim 1 ,further wherein the purified recombinant I2S contains, on average, at least 16 sialic acids per molecule.67.-. (canceled)8. The composition of claim 1 ,further wherein the purified recombinant I2S contains at least 10% bis-phosphorylated oligosaccharides per enzyme.9. (canceled)10. The composition of claim 1 ,further wherein the purified I2S is characterized with a glycan map comprising seven or fewer peak groups selected from the peak groups indicative of neutral (peak group 1), mono-sialylated (peak group 2), di-sialylated (peak group 3), monophosphorylated (peak group 4), tri-sialylated (peak group 5), tetra-sialylated (peak group 6), or diphosphorylated (peak group 7) I2S protein.1112.-. (canceled)13. A composition comprising purified recombinant iduronate-2-sulfatase (I2S) having an amino acid sequence at least 70% identical to SEQ ID NO:1 claim 1 , wherein the purified recombinant I2S comprises greater than about 85% conversion of the cysteine residue corresponding to Cys59 of SEQ ID NO:1 to Cα-formylglycine (FGly).1420.-. (canceled)21. A formulation comprising the composition of and a physiologically ...

Подробнее
Номер записи: 97
02-01-2014 дата публикации

METHOD OF PRODUCING RECOMBINANT IDURONATE-2-SULFATASE

Номер: US20140004097A1
Автор: Boldog Ferenc, Zhang Chun
Принадлежит:

The present invention provides, among other things, methods and compositions for large-scale production of recombinant I2S protein using suspension culture of mammalian cells in serum-free medium. In particular, the present invention uses mammalian cells co-express a recombinant I2S protein and a formylglycine generating enzyme (FGE). 1. A method for large-scale production of recombinant iduronate-2-sulfatase (I2S) protein in mammalian cells , comprising culturing mammalian cells co-expressing a recombinant I2S protein and a formylglycine generating enzyme (FGE) in suspension in a large-scale culture vessel containing medium lacking serum.2. The method of claim 1 , wherein the cells claim 1 , on average claim 1 , produce the recombinant I2S protein at a specific productivity rate of greater than about 15 picogram/cell/day and further wherein the produced recombinant I2S protein claim 1 , on average claim 1 , comprises at least about 60% conversion of the cysteine residue corresponding to Cys59 of human I2S protein to C-formylglycine.3. The method of claim 1 , wherein the culturing step comprises a perfusion process.47.-. (canceled)8. The method of claim 1 , wherein the produced recombinant I2S protein claim 1 , on average claim 1 , comprises at least about 70% conversion of the cysteine residue corresponding to Cys59 of human I2S protein to C-formylglycine.910.-. (canceled)11. The method of claim 1 , wherein the mammalian cells are human cells.12. The method of claim 1 , wherein the mammalian cells are CHO cells.13. The method of claim 1 , wherein the large-scale culture vessel is a bioreactor.1417.-. (canceled)18. The method of claim 1 , wherein the medium comprises at least one redox-modulator selected from the group consisting of glutathione claim 1 , glucose-6-phosphate claim 1 , carnosine claim 1 , carnosol claim 1 , sulforaphane claim 1 , tocopherol claim 1 , ascorbate claim 1 , dehydroascorbate claim 1 , selenium claim 1 , 2-mercaptoenthanol claim 1 , N- ...

Подробнее
Номер записи: 98
02-01-2014 дата публикации

CELLS FOR PRODUCING RECOMBINANT IDURONATE-2-SULFATASE

Номер: US20140004593A1
Автор: Boldog Ferenc, Heartlein Michael
Принадлежит:

The present invention provides, among other things, methods and compositions for production of recombinant I2S protein with improved potency and activity using cells co-express I2S and FGE protein. In some embodiments, cells according to the present invention are engineered to simultaneously over-express recombinant I2S and FGE proteins. Cells according to the invention are adaptable to various cell culture conditions. In some embodiments, cells of the present invention adaptable to a large-scale suspension serum-free culture. 1. A cell comprisinga first nucleic acid encoding an iduronate-2-sulfatase (I2S) protein comprising an amino acid sequence at least 70% identical to SEQ ID NO:1; anda second nucleic acid encoding a formylglycine generating enzyme (FGE) protein comprising an amino acid sequence at least 70% identical to SEQ ID NO:5,wherein the first and/or the second nucleic acid are exogenous and wherein the cell, once cultivated under a cell culture condition, produces I2S protein comprising at least about 70% conversion of the cysteine residue corresponding to Cys59 of SEQ ID NO:1 to Cα-formylglycine (FGly).2. A cell comprisinga first nucleic acid encoding an iduronate-2-sulfatase (I2S) protein comprising an amino acid sequence at least 70% identical to SEQ ID NO:1; anda second nucleic acid encoding a formylglycine generating enzyme (FGE) protein comprising an amino acid sequence at least 70% identical to SEQ ID NO:5,wherein the first and/or the second nucleic acid are exogenous and wherein the cell, once cultivated under a cell culture condition, produces I2S protein comprising at least about 60% conversion of the cysteine residue corresponding to Cys59 of SEQ ID NO:1 to Cα-formylglycine (FGly) and at a specific productivity rate of great than about 30 picogram/cell/day.3. The cell of claim 1 , wherein the cell claim 1 , once cultivated under a cell culture condition claim 1 , produces the I2S protein comprising at least about 80% conversion of the cysteine ...

Подробнее
Номер записи: 99
02-01-2014 дата публикации

Pectate Lyase Variants

Номер: US20140005093A1
Автор: Carsten Andersen, Henrik Frisner, Katja Salomon Johansen, Mads Eskelund BJØRNVAD, Sanne Schoeder Glad, Thomas Thisted, Torben Vedel Borchert
Принадлежит: Novozymes AS

The present invention relates to pectate lyase variants exhibiting alterations relative to a parent enzyme exhibiting pectate lyase activity as its major enzymatic activity; to a method of producing such enzymes; and to methods for using such enzymes in the textile, detergent and cellulose fiber processing industries. Compared to the parent enzyme, the pectate lyase variants of the present invention exhibit improved stability in detergents.

Подробнее
Номер записи: 100