Filamentous Fungal Double-Mutant Host Cells
AssigneeNovozymes A/S
InventorHiroaki Udagawa
Life of this patent
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Abstract
The present invention relates to mutant filamentous fungal host cells producing a polypeptide of interest, wherein two or more polynucleotides encoding two or more polypeptides involved in alpha-glucan synthesis are inactivated; methods of polypeptide production in said mutants as well as methods of constructing such mutants.
Claims19
A mutant filamentous fungal host cell producing a polypeptide of interest, wherein two or more polynucleotides encoding two or more polypeptides involved in alpha-glucan synthesis are inactivated, and wherein said two or more polypeptides are selected from the group consisting of:
- (a) a polypeptide involved in alpha-glucan synthesis comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of the Aspergillus niger AgsA polypeptide of SEQ ID NO:
- 3 and a polypeptide involved in alpha-glucan synthesis comprising an amino acid sequence having at least 70% sequence identity with the Aspergillus niger AgsE polypeptide of SEQ ID NO:6;
- (b) a polypeptide involved in alpha-glucan synthesis encoded by:
- (i) a genomic polynucleotide comprising a nucleotide sequence having a least 70% sequence identity with the Aspergillus niger agsA genomic nucleotide sequence in SEQ ID NO:
- 1 and a polypeptide involved in alpha-glucan synthesis encoded by a genomic polynucleotide comprising a nucleotide sequence having a least 70% sequence identity with the Aspergillus niger agsE genomic nucleotide sequence in SEQ ID NO:4, or
- (ii) the cDNA nucleotide sequences of (i) shown in SEQ ID NO:
- 2 and SEQ ID NO:5, respectively;
- an
- (c) two polypeptides involved in alpha-glucan synthesis encoded by polynucleotides that hybridize under medium stringency conditions with:
- (I) the Aspergillus niger agsA genomic nucleotide sequence in SEQ ID NO:
- 1 and the Aspergillus niger agsE genomic nucleotide sequence in SEQ ID NO:4, respectively;
- (II) the cDNA nucleotide sequences of (I) shown in SEQ ID NO:2 and SEQ ID NO:5, respectively, or
- (III) the full-length complements of (I) or (II).
A method of constructing a mutated filamentous fungal host cell, said method comprising inactivating two or more polynucleotides in a filamentous fungal host cell encoding two or more polypeptides, respectively, selected from the group consisting of:
- (a) a polypeptide involved in alpha-glucan synthesis comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence of the Aspergillus niger AgsA polypeptide of SEQ ID NO:
- 3 and a polypeptide involved in alpha-glucan synthesis comprising an amino acid sequence having at least 70% sequence identity with the Aspergillus niger AgsE polypeptide of SEQ ID NO:6;
- (b) a polypeptide involved in alpha-glucan synthesis encoded by:
- (i) a genomic polynucleotide comprising a nucleotide sequence having a least 70% sequence identity with the Aspergillus niger agsA genomic nucleotide sequence in SEQ ID NO:
- 1 and a polypeptide involved in alpha-glucan synthesis encoded by a genomic polynucleotide comprising a nucleotide sequence having a least 70% sequence identity with the Aspergillus niger agsE genomic nucleotide sequence in SEQ ID NO:4, or
- (ii) the cDNA nucleotide sequences of (i) shown in SEQ ID NO:
- 2 and SEQ ID NO:5, respectively;
- and
- (c) two polypeptides involved in alpha-glucan synthesis encoded by polynucleotides that hybridize under medium stringency conditions with:
- (I) the Aspergillus niger agsA genomic nucleotide sequence in SEQ ID NO:
- 1 and the Aspergillus niger agsE genomic nucleotide sequence in SEQ ID NO:4, respectively;
- (II) the cDNA nucleotide sequences of (I) shown in SEQ ID NO:2 and SEQ ID NO:5, respectively, or
- (III) the full-length complements of (I) or (II).
Dependencies are inferred from the claim text.