Functional expression of keratinase (kerA) gene from Bacillus licheniformis in Pichia pastoris |
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Authors: | Jesus M Porres Maria J Benito Xin Gen Lei |
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Institution: | (1) Department of Animal Science, Cornell University, Ithaca, NY 14853, USA;(2) Present address: Facultad de Veterinaria, Higiene de los Alimentos, Céceres, 10071, Spain |
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Abstract: | A 1.2 kb DNA fragment coding for the pro-peptide and mature keratinase from Bacillus licheniformis PWD-1 (kerA) was cloned into vectors pPICZ A and pGAPZ A for extracellular expression in the methylotrophic yeast, Pichia pastoris. Recombinant keratinase was secreted by the pPICZ A-kerA transformants 24 h after methanol induction of shake-flask cultures, and reached a final yield of 124 mg l–1 (285 U ml–1) 144 h after the induction. The recombinant keratinase was glycosylated ( 39 kDa), and was optimal between pH 8.5–9.5 and between 55 °C –60 °C using azokeratin as substrate. The enzyme degraded bovine serum albumin, collagen, and soy protein concentrate. In conclusion, P. pastoris can be used as an efficient host to express keratinase for nutritional and environmental applications. |
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Keywords: | Bacillus licheniformis functional expression keratinase Pichia pastoris |
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