重组大肠杆菌S3-2产羰基还原酶发酵工艺优化及动力学研究  

Optimization in Fermentation Process and Kinetics of Carbonyl Reductase Production by Recombinant Escherichia coli S3-2

作　　者：田俊 龚大春 饶振辉 TIAN Jun;GONG Dachun;RAO Zhenhui(College of Biological&Pharmaceutical Sciences,China Three Gorges University,Yichang 443002,China;The Second People′s Hospital Affiliated to China Three Gorges University,Yichang 443099,China;Angel Yeast Co.,Ltd.,Yichang 443003,China)

机构地区：[1]三峡大学生物与制药学院,湖北宜昌443002 [2]三峡大学附属第二人民医院,湖北宜昌443099 [3]安琪酵母股份有限公司,湖北宜昌443003

出　　处：《化学与生物工程》2024年第5期25-30,43,共7页Chemistry & Bioengineering

基　　金：国家自然科学基金项目(21776162)。

摘　　要：以重组大肠杆菌S3-2为发酵菌株,通过单因素实验和正交实验对重组大肠杆菌S3-2产羰基还原酶的1 L发酵罐发酵工艺进行优化,并通过Logistic方程和Luedeking-Piret方程对重组大肠杆菌S3-2分批发酵产羰基还原酶的动力学过程进行模拟。结果表明,重组大肠杆菌S3-2的最佳发酵工艺为:IPTG在OD600值为1.0时诱导、IPTG终浓度为0.30 mmol·L^(-1)、种子液OD600值为3.0时接种、诱导温度为26℃,在此条件下,菌体细胞干重为1.586 g·L^(-1)、羰基还原酶酶活为0.712 U·mL^(-1);菌体生长、羰基还原酶酶活及甘油消耗的动力学模型的拟合度分别为0.9883、0.9917和0.9807,说明该模型对重组大肠杆菌S3-2的发酵动力学模型拟合良好,为后续中试放大研究提供了理论依据。Using the recombinant Escherichia coli S3-2 as a fermentation strain,we optimized the fermentation process of the recombinant E.coli S3-2 for the production of carbonyl reductase in a 1 L fermentor through single-factor experiments and orthogonal experiments.Moreover,we used Logistic equation and Luedeking-Piret equation to simulate the kinetic model of carbonyl reductase production by batch fermentation of recombinant E.coli S3-2.The results show that the optimal fermentation process of recombinant E.coli S3-2 is determined as follows:IPTG induction at OD 600 value of 1.0,IPTG final concentration of 0.30 mmol·L^(-1),inoculation at OD 600 value of 3.0,and induction temperature of 26℃.Under above conditions,the dry weight of the strain is 1.586 g·L^(-1),and the activity of carbonyl reductase is 0.712 U·mL^(-1).The fitting degrees of the kinetic models for strain growth,carbonyl reductase activity,and glycerol consumption are 0.9883,0.9917,and 0.9807,respectively,indicating that the model has a good fit for the fermentation kinetic model of recombinant E.coli S3-2.This study provides a theoretical basis for the subsequent pilot scale-up study.

关 键 词：重组大肠杆菌S3-2 羰基还原酶 发酵工艺优化 正交实验 动力学模型 

分 类 号：TQ920.6[轻工技术与工程—发酵工程] Q814[生物学—生物工程]