Proteases and methods for producing them
AssigneeNovozymes A/S
InventorSoren Flensted Lassen
Life of this patent
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Abstract
A secreted mature polypeptide derived from an S2A or S1E protease is described which after maturation has protease activity, which polypeptide when expressed and before maturation comprises a heterologous pro-region. Further, a method for producing polypeptide is described including the step of cultivating a recombinant host cell including a polynucleotide encoding a secreted mature polypeptide which after maturation has protease activity, which polypeptide when expressed and before maturation includes a heterologous pro-region derived from an S2A or S1E protease to produce a supermatant including the polypeptide. The polypeptide may also be recovered.
Claims7
A method for producing a secreted, mature polypeptide, the method comprising:
- (a) cultivating a recombinant host cell comprising a polynucleotide encoding a protein comprising:
- (i) a heterologous pro-region comprising a sequence having at least 90% sequence identity to SEQ ID NO:30 and having activity to enhance expression of the protein,
- (ii) a signal peptide comprising a sequence encoded by a polynucleotide having at least 90% sequence identity to residues 1-81 of SEQ ID NO:2, and
- (iii) an amino acid sequence having at least 92% sequence identity to residues 166-353 of SEQ ID NO:43 and that has protease activity, and
- (b) culturing the host cell so as to produce a culture supernatant comprising the secreted, mature polypeptide, said secreted mature polypeptide lacking the heterologous pro-region and the signal peptide, and comprising the sequence having at least 90% sequence identity to residues 166-353 of SEQ ID NO:43 , wherein said sequence has protease activity, and
- (c) recovering the secreted, mature polypeptide.
An isolated polynucleotide encoding a polypeptide comprising;
- (i) a heterologous pro-region comprising a sequence having at least 90% sequence identity to SEQ ID NO:30 and having activity to enhance expression of the protein,
- (ii) a signal peptide comprising a sequence encoded by a polynucleotide having at least 90% sequence identity to residues 1-81 of SEQ ID NO:2, and
- (iii) an amino acid sequence having at least 92% sequence identity to residues 166-353 of SEQ ID NO:43 and that has protease activity.
A method for producing a secreted mature polypeptide, the method comprising:
- (a) cultivating a recombinant host cell comprising a polynucleotide encoding a protein comprising:
- (i) a heterologous pro-region comprising a sequence having at least 92% sequence identity to SEQ ID NO:30 and having activity to enhance expression of the protein,
- (ii) a signal peptide comprising a sequence encoded by a polynucleotide having at least 90% sequence identity to residues 1-81 of SEQ ID NO:2, and
- (iii) an amino acid sequence having at least 98% sequence identity to resides 166-353 of SEQ ID NO:43 and that has protease activity, and
- (b) culturing the host cell so as to produce a culture supernatant comprising the secreted, mature polypeptide, said secreted mature polypeptide lacking the heterologous pro-region and the signal peptide, and comprising the sequence having at least 98% sequence identity to residues 166-353 of SEQ ID NO:43, wherein said sequence has protease activity.
Dependencies are inferred from the claim text.