Overexpression of Bacillus subtilis Keratinase Gene (kerC) in Pichia pastoris

doi:10.7506/spkx1002-6630-201309053

Abstract

Abstract:

A stable and highly effective expression system was constructed for the expression of keratinase gene (kerC)
in Pichia pastoris. kerC gene was cloned from Bacillus subtilis B-3 by PCR, inserted into the expression vector pPICZαA and
transformed into Pichia pastoris X-33. The results showed that the kerC gene was over expressed successfully in the eukaryotic
expression system. The maximum keratinase yield of the recombinant strains reached 32.31 U/mL in the culture supernatant
grown for 108 h by 1% methanol induction, a 11.15-fold increase in comparison to that obtained with the prokaryotic expression
strain BL21-pET-kerC. The optimal temperature and pH of the recombinant keratinase were 60 ℃ and 7.5, respectively. It was
strongly stimulated by Co2+, Fe2+ and Ca2+ despite being completely inhibited by Fe3+. The recombinant keratinase with a
molecular mass of 31 kD was purified as determined by SDS-PAGE.

Key words: Bacillus subtilis, keratinase, kerC gene, Pichia pastoris, overexpression

CLC Number:

Q814

HAN Xue-yi，TANG Zi-zhong，WANG Li-hua，CHEN Hui*. Overexpression of Bacillus subtilis Keratinase Gene (kerC) in Pichia pastoris[J]. FOOD SCIENCE, 2013, 34(9): 262-266.

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