Methods For Enhancing The Degradation Or Conversion Of Cellulosic Material

Patent metadata

Field	Value
Publication number	US-2016304847-A1
Application number	US-201615199061-A
Country	US
Kind code	A1
Filing date	Jun 30, 2016
Priority date	Mar 31, 2011
Publication date	Oct 20, 2016
Grant date	—

Abstract

The present invention relates to methods for degrading or converting a cellulosic material and for producing a substance from a cellulosic material.

First claim

1 - 20 . (canceled)

21 . An isolated recombinant host cell transformed with a nucleic acid construct comprising a polynucleotide encoding a GH61 polypeptide having cellulolytic enhancing activity, wherein the polynucleotide is operably linked to one or more control sequences that direct the production of the GH61 polypeptide, and wherein the GH61 polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
(a) a GH61 polypeptide having at least 90% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2; (b) a GH61 polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the full-length complement of nucleotides 64 to 1018 or nucleotides 64-101 and 160-1018 of the polynucleotide of SEQ ID NO: 1, or (ii) the cDNA of (i), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.; (c) a GH61 polypeptide encoded by (i) a polynucleotide having at least 90% sequence identity to nucleotides 64 to 1018 or nucleotides 64-101 and 160-1018 of the polynucleotide of SEQ ID NO: 1, or (ii) the cDNA of (i); and (d) a fragment of the GH61 polypeptide of (a), (b), or (c), wherein the fragment has cellulolytic enhancing activity.

22 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity has at least 90% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2.

23 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity has at least 91% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2.

24 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity has at least 92% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2.

25 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity has at least 93% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2.

26 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity has at least 94% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2.

27 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity has at least 95% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2.

28 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity has at least 96% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2.

29 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity has at least 97% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2.

30 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity has at least 98% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2.

31 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity has at least 99% sequence identity to amino acids 22 to 320 of SEQ ID NO: 2.

32 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity comprises (i) the GH61 polypeptide of SEQ ID NO: 2; (ii) amino acids 22 to 320 of the GH61 polypeptide of SEQ ID NO: 2; or (iii) a fragment of (i) or (ii) having cellulolytic enhancing activity.

33 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity is encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the full-length complement of nucleotides 64 to 1018 or nucleotides 64-101 and 160-1018 of the polynucleotide of SEQ ID NO: 1, or (ii) the cDNA of (i), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C.

34 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity is encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the full-length complement of nucleotides 64 to 1018 or nucleotides 64-101 and 160-1018 of the polynucleotide of SEQ ID NO: 1, or (ii) the cDNA of (i), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.

35 . The recombinant host cell of claim 21 , wherein the GH61 polypeptide having cellulolytic enhancing activity is encoded by (i) a polynucleotide comprising SEQ ID NO: 1, (ii) a polynucleotide comprising nucleotides 64 to 1018 or nucleotides 64-101 and 160-1018 of the polynucleotide of SEQ ID NO: 1, or (iii) the cDNA of (i) or (ii).

36 . The recombinant host cell of claim 21 , wherein one or more of the control sequences is heterologous to the polynucleotide encoding the GH61 polypeptide having cellulolytic enhancing activity.

37 . A method of producing a GH61 polypeptide having cellulolytic enhancing activity, comprising cultivating the recombinant host cell of claim 21 under conditions conducive for production of the GH61 polypeptide.

Assignees

• Novozymes Inc

Inventors

• Schnorr Kirk
• Kramer Randall

Classifications

• C12Y302/01

  Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1) · CPC title

• C12N9/2402 Primary

  hydrolysing O- and S- glycosyl compounds (3.2.1) · CPC title

• C12P19/02

  Monosaccharides (2-ketogulonic acid C12P7/60) · CPC title

• C12N9/0071

  acting on paired donors with incorporation of molecular oxygen (1.14) · CPC title

• C12P19/12

  Disaccharides · CPC title