毕赤酵母GS115中N-乙酰转移酶在大肠杆菌中的克隆表达与性质研究

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

生物技术通报 ›› 2015, Vol. 31 ›› Issue (11): 179-185.doi: 10.13560/j.cnki.biotech.bull.1985.2015.11.023

毕赤酵母GS115中N-乙酰转移酶在大肠杆菌中的克隆表达与性质研究

朱海峰^1,2, 吴丹^1,2, 吴敬^1,2

1.江南大学食品科学与技术国家重点实验室，无锡 214122；
2.江南大学生物工程学院工业生物技术教育部重点实验室江南大学，无锡 214122

收稿日期:2015-03-05 出版日期:2015-11-26 发布日期:2015-11-26
作者简介:朱海峰，男，硕士，研究方向: 发酵工程；E-mail: zhuhaifeng009@126.com

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

Zhu Haifeng^1，2, Wu Dan^1，2, Wu Jing^1，2

1. State Key Laboratory of Food Science and Technology，Jiangnan University，Wuxi 214122；
2. School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education，Jiangnan University，Wuxi 214122

Received:2015-03-05 Published:2015-11-26 Online:2015-11-26

摘要/Abstract

摘要： 为了研究P. pastoris Mpr1的生理特性，在E. coli JM109胞内中成功表达来源于P. pastoris GS115的Mpr1酶，并利用响应面分析法对诱导温度、IPTG诱导浓度、起始诱导OD进行发酵优化，酶活达到(610.3±9.5)mU/mL。酶学性质显示，Mpr1酶的最适反应pH范围为7.0-7.5，最适反应温度是30℃。通过分析重组表达Mpr1菌株和对照菌株的培养过程，显示重组菌株的生长能力显著增强，原因是Mpr1降低了细胞内的ROS水平。

关键词: N-乙酰转移酶, 大肠杆菌, 毕赤酵母GS115, 响应面分析法, 发酵优化

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

朱海峰, 吴丹, 吴敬. 毕赤酵母GS115中N-乙酰转移酶在大肠杆菌中的克隆表达与性质研究[J]. 生物技术通报, 2015, 31(11): 179-185.

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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毕赤酵母GS115中N-乙酰转移酶在大肠杆菌中的克隆表达与性质研究

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

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