High-level production of glucose oxidase by recombinant Pichia pastoris using a combined strategy.

Qingxuan MU; Meirong HU; Fei Chen; Xianzhang Jiang; Yong Tao; Jianzhong Huang

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High-level production of glucose oxidase by recombinant Pichia pastoris using a combined strategy.

Author: Qingxuan MU ¹ ; Meirong HU ² ; Fei CHEN ¹ ; Xianzhang JIANG ¹ ; Yong TAO ² ; Jianzhong HUANG ¹
Author Information

1. College of Life Sciences, Fujian Normal University, Fuzhou 350108, Fujian, China.
2. Institute of Microbiology, Chinese Academy of Scienses, Beijing 100101, China.
Publication Type:Journal Article
Keywords: Pichia pastoris; Vitreoscialla hemoglobin; glucose oxidase; protein disulfide isomerase; sorbitol
MeSH: Bioreactors; Fermentation; Glucose Oxidase; biosynthesis; Methanol; Pichia; metabolism; Recombinant Proteins; biosynthesis; Sorbitol
From: Chinese Journal of Biotechnology 2016;32(7):986-990
CountryChina
Language:Chinese
Abstract: To enhance the production of glucose oxidase by recombinant Pichia pastoris, two strategies were developed, which were namely co-feeding of methanol and sorbitol and co-expressing of the protein disulfide isomerase (PDI) and Vitreoscialla hemoglobin (VHb). The volumetric activity reached 456 U/mL by using the strain X33/pPIC9k-GOD, in 5 liter fermentator, with the co-feeding of methanol and sorbitol, it was 0.2 fold higher than that only feeding by methanol. The improved strain was obtained by co-expressing PDI-VHb with GOD. While fermented in a 5 liter fermentator by feeding methanol and sorbitol, the activity of the improved strain reached 716 U/mL with a yield of 7 400 mg/L total soluble protein concentration. These results indicated that heterologous protein expression level can be enhanced by optimizing fermentation condition and co-expression molecular chaperon in Pichia pastoris.