Patent ID: 8715985

Claim:
A method for production of a recombinant C. histolyticum collagenase comprising: a) designing an optimized nucleotide sequence for expression of said recombinant C. histolyticum collagenase, wherein said optimized nucleotide sequence is SEQ ID NO: 1 or SEQ ID NO: 3; b) introducing, in an inducible expression vector, a nucleotide sequence coding for a fusion protein, wherein said nucleotide sequence comprises said optimized sequence fused to a purification tag coding sequence and a soluble polypeptide coding sequence, and wherein said nucleotide sequence is operatively linked to an inducible promoter sequence, a transcription start sequence, and a termination sequence; c) transforming a bacterial strain defective in expression of endogenous proteases with said expression vector; d) culturing said transformed bacterial strain at a temperature in a range from 28° C. to 32° C.; e) inducing expression of said fusion protein by adding a suitable inductor in said bacterial strain; f) extracting the fusion protein from the induced bacterial strain; and g) purifying said fusion protein; wherein said method provides a yield higher than 140 mg/l of culture of said recombinant C. histolyticum collagenase in soluble and enzymatically active form.