Cellulase variants

Details Cellulase variants Cellulase variants

2011-08-09

2011-08-09

Pak, Yong D (Department: 1652)

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

C435S440000, C435S252300, C435S320100, C435S069100, C435S071100, C435S004000, C435S006120, C536S023200

Reexamination Certificate

active

07993898

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) fromHumicola insolensknown 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; andg. 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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