β-glucosidase variants
First Claim
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1. A method of producing a variant β
- -xylosidase polypeptide with improved thermoactivity, comprising;
a) identifying a first performance sensitive position (PSP) in a target β
-xylosidase polypeptide, comprising the steps of;
i) aligning the primary sequence of the target β
-xylosidase polypeptide with one or both of a GH3 consensus sequence SEQ ID NO;
53 or portion thereof and a GH3-C consensus sequence SEQ ID NO;
54 or portion thereof; and
ii) identifying a position in the target β
-xylosidase polypeptide that corresponds in the alignment to a PSP in the one or more a β
-glucosidase polypeptides, wherein the position so identified is a PSP; and
b) expressing a variant β
-xylosidase polypeptide in which the residue at the first performance sensitive position is replaced with a thermoactivity enhancing residue, wherein the variant β
-xylosidase polypeptide has greater thermoactivity than the target β
-xylosidase polypeptide.
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Abstract
The invention relates to recombinantly produced β-glucosidase variants with enhanced thermoactivity compared to naturally occurring proteins. The invention also provides methods for producing a variant β-glucosidase polypeptide with improved thermoactivity by identifying performance sensitive positions in a target β-glucosidase polypeptide and substituting the residue at that position with a thermoactivity enhancing residue.
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Citations
11 Claims
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1. A method of producing a variant β
- -xylosidase polypeptide with improved thermoactivity, comprising;
a) identifying a first performance sensitive position (PSP) in a target β
-xylosidase polypeptide, comprising the steps of;i) aligning the primary sequence of the target β
-xylosidase polypeptide with one or both of a GH3 consensus sequence SEQ ID NO;
53 or portion thereof and a GH3-C consensus sequence SEQ ID NO;
54 or portion thereof; andii) identifying a position in the target β
-xylosidase polypeptide that corresponds in the alignment to a PSP in the one or more a β
-glucosidase polypeptides, wherein the position so identified is a PSP; andb) expressing a variant β
-xylosidase polypeptide in which the residue at the first performance sensitive position is replaced with a thermoactivity enhancing residue, wherein the variant β
-xylosidase polypeptide has greater thermoactivity than the target β
-xylosidase polypeptide. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
- -xylosidase polypeptide with improved thermoactivity, comprising;
Specification