Cloning of Gene for a Glucose Oxidase from Penicillium notatum and Its Enzymatic Properties

doi:10.13560/j.cnki.biotech.bull.1985.2016.07.023

Abstract

Abstract: The purpose of this work is to clone the gene of glucose oxidase（GOD）from Penicillium notatum,which was then in heterologous expression in Pichia pastoris,and the enzymatic properties of purified proteins were studied. GOD gene was cloned from genome DNA of P. notatum No. 8312 strain using PCR technique,the gene was ligated to the vector pMD-AOX and then expressed in strain X33 of P. pastoris,and finally the enzymatic properties of the purified proteins were analyzed. As results,the strain X33-GOD highly expressed the GOD with activity. Its activity of GOD in cultured supernatant reached 496 U/mL and the specific activity was 123.0 U/mg at 30℃ and pH6.5. The optimal temperature and pH of recombinant GOD was 40-45℃ and 6.0 respectively. The result of stability of the enzyme showed that the enzyme was stable when the temperature under 50℃ and the pH3.5-7.0,and it can be activated by 1 mmol/L Zn²⁺ and obvious inhibited by Ag⁺. The recombinant engineering P. pastoris strain with high-yield GOD has a higher fermentation activity and specific activity compared with GOD from P. notatum.

Key words: glucose oxidase, Penicillium notatum, recombinant expression

GAO Qing-hua, HU Mei-rong, WU Fang-tong, TAO Yong, WANG Yun-peng, LUO Tong-yang, HU Chang-ying. Cloning of Gene for a Glucose Oxidase from Penicillium notatum and Its Enzymatic Properties[J]. Biotechnology Bulletin, 2016, 32(7): 152-159.

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