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1. WO2011121768 - NOVEL CELLULASE GENE

Publication Number WO/2011/121768
Publication Date 06.10.2011
International Application No. PCT/JP2010/055897
International Filing Date 31.03.2010
IPC
C12N 9/42 2006.1
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
14Hydrolases (3.)
24acting on glycosyl compounds (3.2)
42acting on beta-1, 4-glucosidic bonds, e.g. cellulase
C12N 1/15 2006.1
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
1Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
14Fungi ; Culture media therefor
15modified by introduction of foreign genetic material
C12N 1/19 2006.1
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NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
1Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
14Fungi ; Culture media therefor
16Yeasts; Culture media therefor
19modified by introduction of foreign genetic material
C12N 1/21 2006.1
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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1Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
20Bacteria; Culture media therefor
21modified by introduction of foreign genetic material
C12N 5/10 2006.1
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
10Cells modified by introduction of foreign genetic material, e.g. virus-transformed cells
C12N 15/09 2006.1
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
CPC
A23K 10/14
AHUMAN NECESSITIES
23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
KFODDER
10Animal feeding-stuffs
10obtained by microbiological or biochemical processes
14Pretreatment of feeding-stuffs with enzymes
A23K 20/189
AHUMAN NECESSITIES
23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
KFODDER
20Accessory food factors for animal feeding-stuffs
10Organic substances
189Enzymes
C12N 9/2437
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NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
14Hydrolases (3)
24acting on glycosyl compounds (3.2)
2402hydrolysing O- and S- glycosyl compounds (3.2.1)
2405Glucanases
2434acting on beta-1,4-glucosidic bonds
2437Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
C12N 9/2445
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9Enzymes; Proenzymes; Compositions thereof
14Hydrolases (3)
24acting on glycosyl compounds (3.2)
2402hydrolysing O- and S- glycosyl compounds (3.2.1)
2405Glucanases
2434acting on beta-1,4-glucosidic bonds
2445Beta-glucosidase (3.2.1.21)
C12P 19/02
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PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
19Preparation of compounds containing saccharide radicals
02Monosaccharides
C12P 19/14
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
19Preparation of compounds containing saccharide radicals
14produced by the action of a carbohydrase ; (EC 3.2.x); , e.g. by alpha-amylase ; , e.g. by cellulase, hemicellulase
Applicants
  • Meiji Seikaファルマ株式会社 Meiji Seika Pharma Co., Ltd. [JP]/[JP] (AllExceptUS)
  • 横山 史和 YOKOYAMA, Fumikazu [JP]/[JP] (UsOnly)
Inventors
  • 横山 史和 YOKOYAMA, Fumikazu
Agents
  • 森田 憲一 MORITA, Kenichi
Priority Data
Publication Language Japanese (ja)
Filing Language Japanese (JA)
Designated States
Title
(EN) NOVEL CELLULASE GENE
(FR) NOUVEAU GÈNE DE CELLULASE
(JA) 新規セルラーゼ遺伝子
Abstract
(EN) An endoglucanase gene or a β-glucosidase gene can be identified by isolating genomic DNA containing a gene for a cellulase that is classified as an endoglucanase or a β-glucosidase from Acremonium cellulolyticus and analyzing the nucleotide sequence for the genomic DNA. Amino acid sequences for known endoglucanases and β-glucosidases are compared with each other extensively, and an amino acid sequence which can be conserved in Acremonium cellulolyticus is found. Based on the information for the amino acid sequence, various primers are designed. Using the various primers thus designed, a PCR is carried out employing genomic DNA or cDNA as a template. As a result, fragments of genes for endoglucanases and genes for β-glucosidases can be obtained. Primers are designed based on the gene fragments, the PCR is continued using the primers to amplify each of a total of nine endoglucanase and β-glucosidase genes, and the nucleotide sequences for the amplification products are analyzed.
(FR) L'invention concerne un gène d'endoglucanase ou un gène de β-glucosidase qui peut être identifié en isolant de l'ADN génomique contenant un gène pour une cellulase qui est classée comme une endoglucanase ou une β-glucosidase provenant d'Acremonium cellulolyticus et en analysant la séquence nucléotidique de l'ADN génomique. Les séquences d'acides aminés pour les endoglucanases et β-glucosidases connues sont comparées les unes par rapport aux autres de manière approfondie et une séquence d'acides aminés qui peut être conservée dans Acremonium cellulolyticus est trouvée. Sur la base des informations sur la séquence d'acides aminés, diverses amorces sont conçues. En utilisant les diverses amorces ainsi conçues, une PCR est réalisée en employant l'ADN génomique ou l'ADNc comme matrice. Ainsi, des fragments de gènes pour les endoglucanases et de gènes pour les β-glucosidases peuvent être obtenus. Les amorces sont conçues en se basant sur les fragments de gène, la PCR est réalisée à l'aide des amorces pour amplifier chacun d'un total de neuf gènes d'endoglucanase et de β-glucosidase et les séquences nucléotidiques des produits d'amplification sont analysées.
(JA)  アクレモニウム・セルロリティカスよりエンドグルカナーゼやβ-グルコシターゼに分類されるセルラーゼ遺伝子を含むゲノムDNAを単離し、その塩基配列を解析することによってエンドグルカナーゼ及びβ-グルコシターゼ遺伝子を特定することを課題とした。 既知のエンドグルカナーゼ及びβ-グルコシターゼのアミノ酸配列を鋭意比較し、アクレモニウム・セルロリティカスにおいても保存されうるアミノ酸配列を見出し、本情報をもとに様々なプライマーを設計した。この様にして設計された様々なプライマーを用いて、ゲノムDNAもしくはcDNAを鋳型にしたPCRを実施した。その結果、エンドグルカナーゼ及びβ-グルコシターゼの遺伝子断片が得られたので、本遺伝子断片を基に、プライマーを設計し、PCRを継続することにより、エンドグルカナーゼ及びβ-グルコシターゼの遺伝子9種を増幅し、塩基配列を解析し本発明を完成するに至った。
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