BGL4 beta-glucosidase and nucleic acids encoding the same
AssigneeGenencor International, Inc.
InventorsNigel Dunn-Coleman, Frits Goedegebuur, Michael Ward, Jian Yao
Life of this patent
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Abstract
The present invention provides a novel β-glucosidase nucleic acid sequence, designated bgl4, and the corresponding BGL4 amino acid sequence. The invention also provides expression vectors and host cells comprising a nucleic acid sequence encoding BGL4, recombinant BGL4 proteins and methods for producing the same.
Claims36
An isolated polynucleotide derived from a fungal source, which polynucleotide comprises a nucleotide sequence encoding an enzyme having β-glucosidase activity.
An isolated polynucleotide selected from the group consisting of:
- (a) a nucleic acid sequence which encodes or is complementary to a sequence which encodes a BGL4 polypeptide having at least 85% sequence identity to the amino acid sequence presented in FIG. 2 (SEQ ID NO:
- 2);
- (b) a nucleic acid sequence which encodes or is complementary to a sequence which encodes a BGL4 polypeptide having at least 90% sequence identity to the amino acid sequence presented in FIG. 2 (SEQ ID NO:
- 2);
- (c) a nucleic acid sequence which encodes or is complementary to a sequence which encodes a BGL4 polypeptide having at least 95% sequence identity to the amino acid sequence presented in FIG. 2;
- (d) a nucleic acid sequence which encodes or is complementary to a sequence which encodes a BGL4 polypeptide having the amino acid sequence presented in FIG. 2;
- (e) a nucleic acid sequence which encodes or is complementary to a sequence which encodes a BGL4 polypeptide having at least 95% sequence identity to the amino acid sequence presented as SEQ ID NO:
- 2;
- (f) a nucleic acid sequence which encodes or is complementary to a sequence which encodes a BGL4 polypeptide having the amino acid sequence presented as SEQ ID NO:
- 2;
- (g) a nucleic acid sequence presented as SEQ ID NO:
- 3, or the complement thereof;
- and
- (h) a nucleic acid sequence that hybridizes, under high stringency conditions to the sequence presented as SEQ ID NO:
- 3, or the complement or a fragment thereof, wherein said isolated polynucleotide encodes a polypeptide having the biological activity of a β-glucosidase
The isolated polynucleotide encoding an enzyme having β-glucosidase activity, wherein the enzyme is derived from a Trichoderma source.
An expression construct including a polynucleotide sequence (i) having at least 85% sequence identity to the amino acid sequence presented in FIG. 2 (SEQ ID NO:
- 2), or (ii) being capable of hybridizing to a probe derived from the nucleotide sequence disclosed in FIG. 2 under conditions of intermediate to high stringency, or (iii) being complementary to a nucleotide sequence having at least 85% sequence identity to the amino acid sequence presented in FIG. 2 (SEQ ID NO:
- 2).
A substantially purified BGL4 polypeptide with the biological activity of a β-glucosidase, comprising a sequence selected from the group consisting of:
- (a) an amino acid sequence having at least 85% sequence identity to the amino acid sequence presented in FIG. 2 (SEQ ID NO:
- 2);
- (b) an amino acid sequence having at least 90% sequence identity to the amino acid sequence presented in FIG. 2 (SEQ ID NO:
- 2);
- (c) an amino acid sequence having at least 95% sequence identity to the amino acid sequence presented in FIG. 2;
- (d) an amino acid sequence presented in FIG. 2;
- (e) an amino acid sequence having at least 95% sequence identity to the amino acid sequence presented as SEQ ID NO:
- 2;
- (f) an amino acid sequence presented as SEQ ID NO:
- 2;
- (g) a substantially purified biologically active fragment of the amino acid sequence presented as SEQ ID NO:
- 2.
A recombinant host cell comprising a deletion or insertion or other alteration in the bgl4 gene which inactivates the gene and prevents BGL4 polypeptide production.
An antisense oligonucleotide complementary to a messenger RNA that encodes a BGL4 polypeptide having the sequence presented as SEQ ID NO:
- 2, wherein upon exposure to a β-glucosidase-producing host cell, said oligonucleotide decreases or inhibits the production of β-glucosidase by said host cell.
A detergent composition, said composition comprising a polypeptide selected from the group consisting of:
- (a) an amino acid sequence having at least 85% sequence identity to the amino acid sequence presented in FIG. 2 (SEQ ID NO:
- 2);
- (b) an amino acid sequence having at least 90% sequence identity to the amino acid sequence presented in FIG. 2 (SEQ ID NO:
- 2);
- (c) an amino acid sequence having at least 95% sequence identity to the amino acid sequence presented in FIG. 2;
- (d) an amino acid sequence presented in FIG. 2;
- (e) an amino acid sequence having at least 95% sequence identity to the amino acid sequence presented as SEQ ID NO:
- 2;
- (f) an amino acid sequence presented as SEQ ID NO:
- 2;
- (g) a substantially purified biologically active fragment of the amino acid sequence presented as SEQ ID NO:
- 2.
A method of expressing a heterologous polypeptide having β-glucosidase activity in an Aspergillus species, comprising:
- (a) Providing a host Aspergillus with an expression vector comprising a polynucleotide encoding a signal sequence linked to a polynucleotide encoding a heterologous β-glucosidase, thereby encoding a chimeric polypeptide;
- (b) Cultivating said host Aspergillus under conditions suitable for said Aspergillus to produce said chimeric polypeptide, wherein said chimeric polypeptide is produced.
A method of producing ethanol, said method comprising the steps of:
- a) contacting a biomass composition with an enzymatic composition comprising β-glucosidase 4 to yield a sugar solution;
- b) adding to the sugar solution a fermentative microorganism;
- and
- c) culturing the fermentative microorganism under conditions sufficient to produce ethanol,
- wherein the biomass composition may be optionally pretreated.
A method of producing ethanol, said method comprising the steps of:
- a) contacting a biomass composition with an enzymatic composition comprising a β-glucosidase 4 and a fermentative microorganism;
- and
- b) culturing the fermentative microorganism under conditions sufficient to produce ethanol,
- wherein the biomass composition may be optionally pretreated.
Dependencies are inferred from the claim text.