Cloning,Expression and Characterization of Enzyme for Mpr1 from P. pastoris GS115 in RecombinantE.coli

doi:10.13560/j.cnki.biotech.bull.1985.2015.11.023

Abstract

Abstract: Due to the nature in P. pastoris methanol metabolism, it suffers much more ROS oxidative stress. There is one Mpr1 enzyme in P. pastoris. It plays significant physiological roles in ROS oxidative stress resistance ability and related research is still blank. For a detailed study about the physiological characteristics of P. pastoris Mpr1, Mpr1 from P. pastoris GS115 had been successfully expressed in E.coli JM109.Fermentation optimization of recombinant cell was studied from induction temperature, IPTG induction concentration, Initial induction OD by using Response Surface Analysis, activity reached(610.3±9.5)mU/mL. Enzymatic properties showed that the optimal pH of Mpr1 was about 7.0 to 7.5, the optimum temperature of Mpr1 was 30℃. In order to explore the nature of Mpr1, PQE30-E.coli JM109 strain and PQE30-Mpr1-E.coli JM109 strain were cultured under the same fermentation conditions in this experiment. The results showed that the growth capacity of the recombinant strain was stronger. The reason is that Mpr1 reduces levels of intracellular ROS.

Key words: N-acetyl transferase, Escherichia coliJM109, P. pastoris GS115, response surface analysis, fermentation optimiza-tion

Zhu Haifeng, Wu Dan, Wu Jing. Cloning,Expression and Characterization of Enzyme for Mpr1 from P. pastoris GS115 in RecombinantE.coli[J]. Biotechnology Bulletin, 2015, 31(11): 179-185.

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