Abstract
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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Zhang, Jh., Wu, D., Chen, J. et al. Enhancing functional expression of β-glucosidase in Pichia pastoris by co-expressing protein disulfide isomerase. Biotechnol Bioproc E 16, 1196–1200 (2011). https://doi.org/10.1007/s12257-011-0136-1
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DOI: https://doi.org/10.1007/s12257-011-0136-1