Cellulase variants
First Claim
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1. A method of producing an endoglucanase variant, comprising introducing a mutation in the amino acid sequence of a parent endoglucanase comprising the amino acid sequence of SEQ ID NO:
- 5 at position 119wherein the mutation is a substitution, insertion or deletion, the endoglucanase variant has endoglucanase activity and the position is numbered according to the amino acid sequence of the cellulase of SEQ ID NO;
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Abstract
The present invention relates to a method for improving the properties of a cellulolytic enzyme by amino acid substitution, deletion or insertion, the method comprising the steps of:
- a. constructing a multiple alignment of at least two amino acid sequences known to have three-dimensional structures similar to endoglucanase V (EGV) from Humicola insolens known from Protein Data Bank entry 4ENG;
- b. constructing a homology-built three-dimensional structure of the cellulolytic enzyme based on the structure of the EGV;
- c. identifying amino acid residue positions present in a distance from the substrate binding cleft of not more than 5 Å;
- d. identifying surface-exposed amino acid residues of the enzyme;
- e. identifying all charged or potentially charged amino acid residue positions of the enzyme;
- f. choosing one or more positions wherein the amino acid residue is to be substituted, deleted or where an insertion is to be provided; and
- g. carrying out the substitution, deletion or insertion by using conventional protein engineering techniques. Also described are cellulase variants obtained by this method.
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Citations
3 Claims
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1. A method of producing an endoglucanase variant, comprising introducing a mutation in the amino acid sequence of a parent endoglucanase comprising the amino acid sequence of SEQ ID NO:
- 5 at position 119
wherein the mutation is a substitution, insertion or deletion, the endoglucanase variant has endoglucanase activity and the position is numbered according to the amino acid sequence of the cellulase of SEQ ID NO;
1. - View Dependent Claims (2, 3)
- 5 at position 119
Specification
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Current AssigneeNovozymes A/S (Novozymes)
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Original AssigneeNovozymes A/S (Novozymes)
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InventorsAndersen, Kim Vilbour, Christiansen, Lars, Damgaard, Bo, Schulein, Martin, Von der Osten, Claus
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Primary Examiner(s)Pak; Yong D
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Application NumberUS11/830,063Publication NumberTime in Patent Office1,506 DaysField of SearchNoneUS Class Current435/209CPC Class CodesC11D 3/38645 containing cellulaseC12N 9/2437 Cellulases (3.2.1.4; 3.2.1....C12Y 302/01004 Cellulase (3.2.1.4), i.e. e...
