Fungus wherein the areA, pepC and/or pepE genes have been inactivated
AssigneeNovo Nordisk A/S
InventorsTove Christensen, Jan Lehmbeck
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Abstract
The present invention relates to fungi, which do not produce proteases. The fungi of the invention are useful as hosts for the production of proteins susceptible of proteolytic degradation by the proteases usually produced, and the invention consequently encompasses processes for the production of proteins of interest in high yields by using the fungi of the invention. The invention also comprises methods for producing such fungi and DNA constructs to be used in these methods.
Claims35
A modified fungus wherein:
- (i) the areA gene no longer expresses a functional AreA activator, and (ii) genes encoding PepC and/or PepE no longer express functional PepC and/or PepE proteases.
A process for producing a desired polypeptide, said process comprising (i) cultivating a fungus in an appropriate growth medium and (ii) recovering said polypeptide from said culture, wherein said fungus comprises a recombinant DNA construct capable of causing expression of said polypeptide or a precursor thereof in said fungus, and wherein said fungus (a) produces lower amounts of functional AreA protein, and (b) produces lower amounts of PepC and/or PepE protein than the wild-type of said fungus.
A DNA sequence coding for the pepC gene from A. oryzae (SEQ ID No. 1).
A PepC protease from A. oryzae (SEQ ID No. 2).
A DNA sequence coding for the pepE gene from A. oryzae (SEQ ID No. 3).
A PepE protease from A. oryzae (SEQ ID No. 4).
Dependencies are inferred from the claim text.