Variant
Disclosed are variants of Humicola grisea CeI7A (CBH1.1), H. jecorina CBH1 variant or S. thermophilium CBH1, nucleic acids encoding the same and methods for producing the same. The variant cellulases have the amino acid sequence of a glycosyl hydrolase of family 7A wherein one or more amino acid residues are substituted.
1. A polynucleotide encoding a cellobiohydrolase comprising (i) an amino acid sequence that is at least 85% identical to SEQ ID NO:10 and (ii) at least one substitution at a position corresponding to T55, Q101, N250, P265, or L288 of SEQ ID NO:10, wherein the substitution at the position corresponding to T55 is T55K and the substitution at the position corresponding to Q101 is Q101Y or Q101H.
2. The polynucleotide of claim 1 , wherein the cellobiohydrolase comprises one or more substitutions selected from the group consisting of N250D, N250E, P265A, P265S, and L288I.
3. A nucleic acid construct comprising the polynucleotide according to claim 1 , operably linked to one or more control sequences.
4. A recombinant expression vector comprising the nucleic acid construct of claim 3 .
5. A recombinant host cell comprising the nucleic acid construct of claim 3 .
6. A method for producing a CBH1 polypeptide, said method comprising:
a) transforming a host cell with the nucleic acid construct of claim 3 ;
b) culturing the host cell under conditions to produce the polypeptide; and
c) recovering the polypeptide.
7. The polynucleotide of claim 1 , wherein the cellobiohydrolase comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:10.
8. The polynucleotide of claim 7 , wherein the cellobiohydrolase comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:10.
9. A method of converting biomass to sugars comprising contacting the biomass with a cellobiohydrolase encoded by the polynucleotide of claim 1 .