Thermal tolerant cellulase fromAcidothermus cellulolyticus
First Claim
1. A composition comprising a thermostable GuxA poly peptide heterologously expressed in an organism other than Acidothermus cellulolyticus, said GuxA poly peptide comprising a first catalytic domain GH6, a second catalytic domain GH 12, a carbohydrate binding domain (CBD) type III, and a carbohydrate binding domain (CBD) type II.
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Abstract
The invention provides a thermal tolerant cellulase that is a member of the glycoside hydrolase family. The invention further discloses this cellulase as GuxA. GuxA has been isolated and characterized from Acidothermus cellulolyticus. The invention further provides recombinant forms of the identified GuxA. Methods of making and using GuxA polypeptides, including fusions, variants, and derivatives, are also disclosed.
37 Citations
57 Claims
- 1. A composition comprising a thermostable GuxA poly peptide heterologously expressed in an organism other than Acidothermus cellulolyticus, said GuxA poly peptide comprising a first catalytic domain GH6, a second catalytic domain GH 12, a carbohydrate binding domain (CBD) type III, and a carbohydrate binding domain (CBD) type II.
- 12. An isolated thermal tolerant GuxA poly peptide having a sequence of SEQ ID NO:
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24. An isolated polypeptide molecule comprising:
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a) a sequence of SEQ ID NO;
4;b) a sequence of SEQ ID NO;
7;c) a sequence of SEQ ID NO;
5;d) a sequence of SEQ ID NO;
8;e) a sequence of SEQ ID NO;
1;
orf) an amino acid sequence having about 90% sequence identity with the amino acid sequence of a), b), c), d), or e) and having at least one domain of glycosyl hydrolase family 6 and glycosyl hydrolase family 12. - View Dependent Claims (31, 32, 33, 34, 35, 36, 37, 38, 39, 45, 46, 47, 48, 49)
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40. A method for assessing the carbohydrate hydrolysis activity of GuxA polypeptide comprising:
- analyzing a carbohydrate hydrolysis in the presence of GuxA polypeptide and a carbohydrate hydrolysis in the absence of GuxA polypeptide on a substrate; and
comparing the carbohydrate hydrolysis in the presence of GuxA polypeptide with the carbohydrate hydrolysis in the absence of GuxA polypeptide. - View Dependent Claims (43)
- analyzing a carbohydrate hydrolysis in the presence of GuxA polypeptide and a carbohydrate hydrolysis in the absence of GuxA polypeptide on a substrate; and
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41. A method for assessing the carbohydrate hydrolysis activity of GuxA polypeptide in the presence of an agent of interest comprising:
- analyzing a carbohydrate hydrolysis in the presence of GuxA polypeptide and a carbohydrate hydrolysis in the presence of GuxA polypeptide and the agent of interest on a substrate; and
comparing the carbohydrate hydrolysis in the GuxA polypeptide treated substrate with the carbohydrate hydrolysis in the GuxA polypeptide treated substrate in the presence of the agent of interest. - View Dependent Claims (42, 44)
- analyzing a carbohydrate hydrolysis in the presence of GuxA polypeptide and a carbohydrate hydrolysis in the presence of GuxA polypeptide and the agent of interest on a substrate; and
- 51. A composition comprising a GuxA-derived peptide, said peptide comprising at least one catalytic domain selected from GH6 and GH 12 domains, and at least one carbohydrate binding domain selected from CBD type II and CBD type III domains.
Specification