Cellobiohydrolase variants and polynucleotides encoding same
The present invention relates to variants of a parent cellobiohydrolase II. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.
1. An isolated variant of a parent cellobiohydrolase II, comprising a substitution at a position corresponding to position 357 of residues 18-481 of SEQ ID NO: 2, wherein the variant has cellobiohydrolase II activity and is selected from the group consisting of:
(a) a polypeptide having at least 95% sequence identity to residues 18-481 of SEQ ID NO: 2;
(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) nucleotides 52-1443 of SEQ ID NO: 1, (ii) the genomic DNA sequence of nucleotides 52-1443 of SEQ ID NO: 1, or (iii) the full-length complementary strand of (i) or (ii); and
(c) a polypeptide encoded by a polynucleotide having at least 99% identity to nucleotides 52-1443 of SEQ ID NO: 1 or the genomic DNA sequence thereof.
2. The variant of claim 1 , which comprises a substitution at a position corresponding to position 357 of the mature polypeptide of SEQ ID NO: 2 with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr or comprises a substitution at position 357 of the mature polypeptide of SEQ ID NO: 2 with Ala, Arg, Asn, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr.
3. The variant of claim 1 , which further comprises a substitution at a position corresponding to position 435 of the mature polypeptide of SEQ ID NO: 2 with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr or further comprises a substitution at position 435 of the mature polypeptide of SEQ ID NO: 2 with Ala, Am, Asn, Asp, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr.
4. An isolated polynucleotide encoding the variant of claim 1 .
5. A method of producing a variant of a parent cellobiohydrolase II, said method comprising:
(a) cultivating an isolated host cell comprising the polynucleotide of claim 4 under conditions suitable for the expression of the variant; and
(b) recovering the variant.
6. A transgenic plant, plant part or plant cell transformed with the isolated polynucleotide of claim 4 .
7. A method for degrading or converting a cellulosic material, said method comprising treating the cellulosic material with an enzyme composition in the presence of the variant of claim 1 .
8. A method for producing a fermentation product, said method comprising:
(a) saccharifying a cellulosic material with an enzyme composition in the presence of the variant of claim 1 ;
(b) fermenting the saccharified cellulosic material with one or more fermenting microorganisms to produce the fermentation product; and
(c) recovering the fermentation product from the fermentation.
9. A method of fermenting a cellulosic material, said method comprising: fermenting the cellulosic material with one or more fermenting microorganisms, wherein the cellulosic material is saccharified with an enzyme composition in the presence of the variant of claim 1 .
10. The variant of claim 2 , wherein the substitution is Asn.
11. The variant of claim 1 , wherein the parent cellobiohydrolase II has at least 95% sequence identity to residues 18-481 of SEQ ID NO:2.
12. The variant of claim 1 , wherein the parent cellobiohydrolase II is encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) nucleotides 52-1443 of SEQ ID NO:1, (ii) the genomic DNA sequence of nucleotides 52-1443 of SEQ ID NO:1, or (iii) the full-length complementary strand of (i) or (ii).
13. The variant of claim 1 , wherein the parent cellobiohydrolase II is encoded by a polynucleotide at least 99% sequence identity to nucleotides 52-1443 of SEQ ID NO:1.
14. The variant of claim 1 , wherein the parent cellobiohydrolase II comprises or consists of residues 18-481 of SEQ ID NO:2, or a fragment thereof having cellobiohydrolase activity.
15. A method of producing the variant of claim 1 , said method comprising:
(a) cultivating a transgenic plant or a plant cell comprising a polynucleotide encoding the variant under conditions conducive for production of the variant; and
(b) recovering the variant.
16. The method of claim 7 , said method further comprising recovering the degraded cellulosic material.
17. The method of claim 9 , wherein the fermenting of the cellulosic material produces a fermentation product.
18. The variant of claim 1 , wherein said variant has increased thermostability relative to the parent.