Fungal cellobiohydrolases
View Patent ↗Disclosed herein are chimeric Cel7A polypeptides useful for producing biofuels from lignocellulosic biomass.
1. A non-naturally occurring, mutated Cel7A polypeptide having increased hydrolase activity when compared to the naturally occurring Cel7A polypeptide from which the non-naturally occurring mutated Cel7A polypeptide was derived; and
wherein said mutated Cel7A polypeptide has an amino acid sequence identity of at least 85% of SEQ ID NO: 2.
2. The non-naturally occurring mutated Cel7A polypeptide of claim 1 wherein the non-naturally occurring mutated Cel7A polypeptide comprises a L371M mutation of SEQ ID NO: 2.
3. The non-naturally occurring mutated Cel7A polypeptide of claim 1 wherein the non-naturally occurring mutated Cel7A polypeptide comprises a D92E mutation of SEQ ID NO: 2.
4. The non-naturally occurring mutated Cel7A polypeptide of claim 1 wherein the non-naturally occurring mutated Cel7A polypeptide comprises a A239V mutation of SEQ ID NO: 2.
5. The non-naturally occurring mutated Cel7A polypeptide of claim 1 wherein the non-naturally occurring mutated Cel7A polypeptide comprises a T481I mutation of SEQ ID NO: 2.
6. The non-naturally occurring mutated Cel7A polypeptide of claim 1 having up to 2.3 times the hydrolase activity when compared to the naturally occurring Cel7A polypeptide from which the non-naturally occurring mutated Cel7A polypeptide was derived.
7. The non-naturally occurring mutated Cel7A polypeptide of claim 1 having up to 2.3 times the hydrolase activity when compared to the naturally occurring Cel7A polypeptide from which the non-naturally occurring mutated Cel7A polypeptide was derived wherein the non-naturally occurring Cel7A polypeptide comprises a L371M mutation of SEQ ID NO: 2.
8. The non-naturally occurring mutated Cel7A polypeptide of claim 1 having up to 2.3 times the hydrolase activity when compared to the naturally occurring Cel7A polypeptide from which the non-naturally occurring mutated Cel7A polypeptide was derived wherein the non-naturally occurring Cel7A polypeptide comprises a D92E mutation of SEQ ID NO: 2.
9. The non-naturally occurring mutated Cel7A polypeptide of claim 1 having up to 2.3 times the hydrolase activity when compared to the naturally occurring Cel7A polypeptide from which the non-naturally occurring mutated Cel7A polypeptide was derived wherein the non-naturally occurring Cel7A polypeptide comprises a A239V mutation of SEQ ID NO: 2.
10. The non-naturally occurring mutated Cel7A polypeptide of claim 1 having up to 2.3 times the hydrolase activity when compared to the naturally occurring Cel7A polypeptide from which the non-naturally occurring mutated Cel7A polypeptide was derived wherein the non-naturally occurring Cel7A polypeptide comprises a T481I mutation of SEQ ID NO: 2.
11. An isolated nucleic acid molecule encoding the non-naturally occurring mutated Cel7A polypeptide of claim 1 .
12. The isolated nucleic acid molecule of claim 11 , further comprising a promoter operably linked to the nucleic acid molecule.
13. The isolated nucleic acid molecule of claim 12 , wherein the promoter allows expression of the nucleic acid in a filamentous fungal host cell.
14. The isolated nucleic acid molecule of claim 13 , wherein the nucleic acid is within an expression vector.
15. A host cell expressing a recombinant polypeptide encoded by the nucleic acid molecule of claim 11 .
16. The host cell of claim 15 , wherein the cell is a fungal cell.