β-Glucosidase variants
AssigneeCodexis, Inc.
InventorsXiyun Zhang, Jie Yang, Dipnath Baidyaroy, Sally Rhiannon Postlethwaite, Attila Andor, Louis Clark, Lisa M. Newman
Life of this patent
Estimating the term…
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.
Claims2
A method of producing a variant Myceliophthora thermophila β-glucosidase polypeptide with improved thermoactivity, comprising:
- a) identifying a first performance sensitive position (PSP) in a target Myceliophthora thermophila β-glucosidase polypeptide, comprising the steps of:
- i) aligning the primary sequence of the target Myceliophthora thermophila β-glucosidase polypeptide with one or both of SEQ ID NO:53 or portion thereof, and SEQ ID NO:54, or portion thereof, and
- ii) identifying a position in the target Myceliophthora thermophila β-glucosidase polypeptide that corresponds in the alignment to a PSP in the one or more β-glucosidase polypeptides, wherein the position so identified is a PSP;
- and
- b) expressing a variant Myceliophthora thermophila β-glucosidase polypeptide in which the residue at the first performance sensitive position is replaced with a thermoactivity enhancing residue,
- wherein the variant Myceliophthora thermophila β-glucosidase polypeptide has greater thermoactivity than the target Myceliophthora thermophila β-glucosidase polypeptide.
A method of producing a variant Myceliophthora thermophila β-glucosidase polypeptide with improved thermoactivity, comprising
- a) identifying a first performance sensitive position (PSP) in a target Myceliophthora thermophila β-glucosidase polypeptide, comprising the steps of:
- i) aligning the primary sequence of the target Myceliophthora thermophila β-glucosidase polypeptide with one or more Myceliophthora thermophila β-glucosidase polypeptides in which performance sensitive positions have been defined, and
- ii) identifying a position in the target Myceliophthora thermophila β-glucosidase polypeptide that corresponds in the alignment to a PSP in the one or more Myceliophthora thermophila β-glucosidase polypeptides, wherein the position so identified is a PSP;
- and
- b) expressing a variant Myceliophthora thermophila β-glucosidase polypeptide in which the residue at the first performance sensitive position is replaced with a thermoactivity enhancing residue,
- wherein the variant Myceliophthora thermophila β-glucosidase polypeptide has greater thermoactivity than the target Myceliophthora thermophila β-glucosidase polypeptide, wherein the step of identifying a PSP comprises aligning the primary sequence of the target Myceliophthora thermophila β-glucosidase polypeptide with at least a portion of SEQ ID NO:
- 1, and wherein the PSP corresponds to a Myceliophthora thermophila Bgl1 PSP at position 350, wherein position 350 is determined relative to SEQ ID NO:
- 1.
Dependencies are inferred from the claim text.