产腈水解酶基因工程菌的产酶诱导条件及培养基优化  

Optimization of culture conditions for production of nitrilase by using recombinant E.coli

作　　者：唐璐敏[1] 薛建萍[1] 

机构地区：[1]上海市农药研究所生物工程中心,上海200032

出　　处：《工业微生物》2016年第5期1-10,共10页Industrial Microbiology

基　　金：国家高技术研究发展计划(863计划);课题编号SS2014AA022106

摘　　要：本文通过对产酶诱导条件及发酵培养基进行优化,成功提高了产腈水解酶基因工程菌E.coli BL21(DE3)-pETNYNit的产酶水平。研究结果显示,最佳发酵培养基为:葡萄糖0.2%、甘油0.7%(v/v)、蛋白胨1.2%、酵母膏0.8%、NaCl0.3%、(NH_4)2SO_40.3%、NH_4Cl0.13%、Na2HPO_4·12H_2O1.04%、KH_2PO_40.39%、MgSO_4·7H_2O0.03%,pH7.2。最佳产酶诱导条件为:发酵4h时加入0.5mmol/LIPTG,然后在28℃、240r/min下诱导腈水解酶基因表达14h^16h。采用优化方案,重组菌产酶水平可提升至0.9~1×105U,与野生菌株的产酶水平相比,提高幅度超过50%。同时重组菌培养仅需24h,培养周期缩短超过50h。In this study,the production of nitrilase by using recombinant strain E. coli BL21（ DE3） – p ETNYNit was improved by optimization of induced conditions and fermentation medium. The results indicated that the optimal medium were as follows： glucose 0. 2%,glycerol 0. 7%（ v/v）,peptone 1. 2%,yeast extract 0. 8%,Na Cl 0. 3%,（ NH_4）2SO_40. 3%,NH_4 Cl and 0. 13%,Na2HPO_4·12H_2O 1. 04%,KH_2PO_40. 39%,Mg SO_4·7H_2O 0. 03%,pH 7. 2; the optimal induced conditions were the following： 0. 5 mmol/L IPTG induced expression after fermented for 4 hours,and then cultured for 14 hours to 16 hours at 28 ℃ and 240 r/min. After optimization,the nitrilase activity of the recombinant strain increased to（ 1 ~ 0. 9） × 10^5 U. Compared with that of the wild strain,the enzyme activity increased by more than50%. At the same time,the culture time of the recombinant strain was about 24 hours,reduced more than 50 hours.

关 键 词：腈水解酶 重组菌 诱导优化 培养基优化 

分 类 号：TQ925[轻工技术与工程—发酵工程]