Enhancement of Glucose Oxidase in Pichia pastoris by Co-expressing Chaperone PDI and Ero1

doi:10.13560/j.cnki.biotech.bull.1985.2017-1095

Abstract

Abstract: The expression of recombinant glucose oxidase（GOD）in Pichia pastoris was enhanced by the sole expression of protein chaperones protein disulfide isomerase（PDI）as well as the co-expression of endoplasmic reticulum oxidoreductase 1（Ero1）and PDI. The pPICZ/PDI and pPICZ/PDI -Ero1expression plasmids were linearized and integrated into the genome of P. pastoris X33/pMD-GOD by electroporation. The positive transformants were screened with double-resistance plate of 250 μg/mL G418 and 50 μg/mL Zeocin. The positive transformants were fermented in tube and 10 L fermenter for analyzing the effects of co-expressing PDI and Ero1-PDI on the expression of GOD. The results revealed that co-expression of PDI and Ero1-PDI increased GOD activity by 29.7% and 100% and their activity reached 476 U/m L and 736 U/m L respectively in 10 L fermenter at 30℃,compared with the control type of original strain. Conclusively,the integration of chaperone PDI and Ero1 enhanced the correct folding of GOD and significantly increased the expression of GOD.

Key words: glucose oxidase, chaperone, protein expression, Pichia pastoris

GAO Qing-hua, DONG Cong, WANG Yue, HU Mei-rong, WANG Qing-qing, WANG Yun-peng, LUO Tong-yang, LIU Lei. Enhancement of Glucose Oxidase in Pichia pastoris by Co-expressing Chaperone PDI and Ero1[J]. Biotechnology Bulletin, 2018, 34(7): 174-179.

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