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β-glucosidase variants

  • US 8,956,845 B2
  • Filed: 05/28/2014
  • Issued: 02/17/2015
  • Est. Priority Date: 06/16/2009
  • Status: Active Grant
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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