产亚硝酸还原酶基因工程菌pET-28a(+)-nir-BL21的培养基优化  

Optimization of Culture Medium for Nitrite Reductase Produced by Genetic Engineering Bacteria E.coli pET-28a(+)-nir-BL21

作　　者：彭鑫 黄燕燕[1] 赵珊 孙丽娜 陈思敏[1] 肖弘毅 刘冬梅[1] PENG Xin;HUANG Yan-yan;ZHAO Shan;SUN Li-na;CHEN Si-min;XIAO Hong-yi;LIU Dong-mei(School of Food Science and Engineering,South China University of Technology,Guangzhou 510640,China)

机构地区：[1]华南理工大学食品科学与工程学院,广东广州510640

出　　处：《食品工业科技》2020年第10期82-88,共7页Science and Technology of Food Industry

基　　金：国家自然科学基金青年科学基金(31771908);广东省自然科学基金(S2011010005679)。

摘　　要：从豆瓣酱中筛选的一株能高效降解亚硝酸盐的蜡样芽孢杆菌(Bacillus cereus)LJ01,将其nir基因克隆至相应的表达载体上,构建了重组菌pET-28a(+)-nir-BL21。为提高基因工程菌pET-28a(+)-nir-BL21的产亚硝酸盐还原酶(NiR)水平,对重组菌pET-28a(+)-nir-BL21的培养基组成通过单因素实验、Plackett-Burman和Box-Behnken试验优化。结果表明,获得最佳培养基为:以液体LB培养基为基础培养基,再添加葡萄糖3.72 g·L^-1、甘油2.63 g·L^-1和细菌学蛋白胨8.70 g·L^-1,活菌数预测值为3.14×10^8 CFU·mL^-1,通过验证得3.02×10^8 CFU·mL^-1,与预测值接近。本实验构建的高产亚硝酸盐还原酶的菌株,将为日后降解食品中的亚硝酸盐进行工业化应用提供理论基础。Bacillus cereus LJ01,isolated from bean paste with efficient nitrite degradation was used to construct the recombinant strain pET-28 a(+)-nir-BL21 with the corresponding expression vectors of nir gene.In order to improve the production of nitrite reductase(NiR)of pET-28 a(+)-nir-BL21,the composition of culture medium was preliminarily optimized.The optimum medium conditions were obtained by single factor test,Plackett-Burman and Box-Behnken experiment.The results showed that the optimum medium were:The liquid LB medium as the base medium,followed by 3.72 g·L^-1 glucose,2.63 g·L^-1 glycerol and 8.70 g·L^-1 peptone.The verified number of viable cells was 3.02×10^8 CFU·mL^-1,which was close to the predicted value(3.14×10^8 CFU·mL^-1).The high-yield nitrite reductase strain constructed in this experiment would provide a theoretical basis for industrial application of nitrite in food degradation in the future.

关 键 词：亚硝酸盐还原酶 重组菌 培养基优化 基因工程 

分 类 号：TS201.3[轻工技术与工程—食品科学]