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AssigneeDyadic International (USA), Inc.
InventorsMark Aaron Emalfarb, Richard Paul Burlingame, Philip Terry Olson, Arkady Panteleimonovich Sinitsyn, Martine Parriche, Jean Christophe Bousson, Christine Marie Pynnonen, Peter Jan Punt, Cornelia Maria Johanna Van Zeijl
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A novel transformation system in the field of filamentous fungal hosts for expressing and secreting heterologous proteins or polypeptides is described. The invention also covers a process for producing large amounts of polypeptide or protein in an economical manner. The system comprises a transformed or transfected fungal strain of the genus Chrysosporium, more particularly of Chrysosporium lucknowense and mutants or derivatives thereof. It also covers transformants containing Chrysosporium coding sequences, as well expression-regulating sequences of Chrysosporium genes. Also provided are novel fungal enzymes and their encoding sequences and expression-regulating sequences.
A mutant Chrysosporium strain comprising a nucleic acid sequence encoding a polypeptide of interest, said nucleic acid sequence being operably linked to an expression-regulating region and optionally a secretion signal sequence, said mutant strain expressing said polypeptide of interest at a higher level than the corresponding non-mutant strain under the same conditions.
A nucleic acid construct comprising a nucleic acid expression-regulatory region derived from Chrysosporium lucknowense, operably linked to a polypeptide-encoding nucleic acid sequence.
A method for producing a mutant Chrysosporium strain according to claim I comprising stably introducing a nucleic acid sequence encoding a heterologous or homologous polypeptide into a Chrysosporium strain, said nucleic acid sequence being operably linked to an expression regulating region.
A protein corresponding to a Chrysosporium glycosyl hydrolase family 7, exhibiting at least 75% amino acid identity as determined by the BLAST algorithm with the amino acid sequence of SEQ ID NO:
A protein corresponding to a Chrysosporium glycosyl hydrolase family 10, exhibiting at least 70% amino acid identity as determined by the BLAST algorithm with the amino acid sequence of SEQ ID NO:2 or a part thereof having at least 20 contiguous amino acids which are identical to the corresponding part of the amino acid sequence 1-383 of SEQ ID NO:2.
A protein corresponding to a Chrysosporium glycosyl hydrolase family 10, exhibiting at least 65% amino acid identity as determined by the BLAST algorithm with the amino acid sequence of SEQ ID NO:2 or a part thereof having at least 20 contiguous amino acids which are identical to the corresponding part of the amino acid sequence 1-383 of SEQ ID NO:2,
A protein corresponding to a Chrysosporium glycosyl hydrolase family 10 and comprising a cellulose-binding domain selected from the group comprising (a) domains having at least 75% amino acid identity with the amino acid sequence 22-53, and (b) domains having at least 20 contiguous amino acids identical to a part of amino acid sequence 22-53 of SEQ ID NO:2.
A fungal glycosyl hydrolase of family 10, comprising a cellulose-binding domain, not derived from Fusarium oxysporum.
A protein corresponding to a Chrysosporium glycosyl hydrolase family 12, exhibiting at least 65% amino acid identity as determined by the BLAST algorithm with the amino acid sequence of SEQ ID NO:3 or a part thereof having at least 20 contiguous amino acids which are identical to the corresponding part of the amino acid sequence 1-247 of SEQ ID NO:3.
A protein corresponding to a Chrysosporium glyceraldehyde phosphate dehydrogenase, exhibiting at least 86% amino acid identity as determined by the BLAST algorithm with the partial amino acid sequence of SEQ ID NO:4 or a part thereof having at least 20 contiguous amino acids which are identical to the corresponding part of the amino acid sequence 1-277 of SEQ ID NO:4.
A protein corresponding to a Chrysosporium glycosyl hydrolase family 45, exhibiting at least 83% amino acid identity as determined by the BLAST algorithm with the amino acid sequence of SEQ ID NO:5 or a part thereof having at least 20 contiguous amino acids which are identical to the corresponding part of the amino acid sequence 1-225 of SEQ ID NO:5.
A protein corresponding to a Chrysosporium glycosyl hydrolase family 6, exhibiting at least 50% amino acid identity as determined by the BLAST algorithm with the amino acid sequence of SEQ ID NO:6 or a part thereof having at least 20 contiguous amino acids which are identical to the corresponding part of the amino acid sequences 1-395 of SEQ ID NO:6.
A nucleic acid sequence comprising at least 70% of the nucleotides contained in the 5′-noncoding region of the nucleic acid sequence of any one of SEQ ID NOS:
A nucleic acid construct comprising a nucleic acid expression-regulatory region derived from Chrysosporium, contained in the 5′-noncoding region of the nucleic acid sequence of any one of SEQ ID No's 1-6, operationally linked to a nucleic acid sequence encoding a polypetide of interest.
An oligonucleotide probe comprising at least 15 contiguous nucleotides of the nucleic acid sequence of any one of SEQ ID No's 1-6, or the complement thereof.
Dependencies are inferred from the claim text.