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KMID : 1094720110160061196
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Biotechnology and Bioprocess Engineering
2011 Volume.16 No. 6 p.1196 ~ p.1200
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Enhancing functional expression of β-glucosidase in Pichia pastoris by co-expressing protein disulfide isomerase
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Zhang Jian-Hong
Wu Dan
Chen Jian
Wu Jing
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Abstract
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The expression of heterologous proteins may exert severe stress on the host cells at different levels. Protein folding and disulfide bond formation were identified as rate-limited steps in recombinant protein secretion in yeast cells. For the production of β-glucosidase in Pichia pastoris, final β-glucosidase activity reached 1,749 U/mL after fermentation optimization in a 3 L bioreactor, while the specific activity decreased from 620 to 467 U/mg, indicating a potential protein misfolding. To solve this problem, protein disulfide isomerase, a chaperone protein which may effectively regulate disulfide bond formation and protein folding, was co-expressed with β-glucosidase. In the co-expression system, a β-glucosidase production level of 2,553 U/mL was achieved and the specific activity of the enzyme reached 721 U/mg, which is 1.54 fold that of the control.
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KEYWORD
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β-glucosidase, fermentation optimization, Pichia pastoris, protein disulfide isomerase, protein folding, specific activity
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