文献类型：期刊文献

中文题名：生物法合成辛伐他汀

英文题名：Biosynthesis of simvastatin

作者：朱丽平[1];任宇红[1];孙常磊[1];周剑平[1];魏东芝[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

年份：2011

卷号：9

期号：5

起止页码：11

中文期刊名：生物加工过程

外文期刊名：Chinese Journal of Bioprocess Engineering

收录：CSTPCD;;CSCD:【CSCD_E2011_2012】；

基金：国家高技术研究发展计划(863计划)资助项目(2008AA02Z204)

语种：中文

中文关键词：酰基转移酶;Monacolin;J;辛伐他汀

外文关键词：acyltransferase ; Monacolin J ; simvastatin

摘要：通过提高E.coli BL21(DE3)/pAW31菌株中的酰基转移酶LovD的表达,并以Monacolin J为底物,催化合成辛伐他汀。考察发酵培养基和发酵条件对酰基转移酶LovD表达的影响;采用SDS-PAGE凝胶电泳法检测酰基转移酶LovD表达情况;并建立酶活测定方法,测定酰基转移酶LovD的实际酶活。通过实验确定酰基转移酶LovD摇瓶发酵的最佳条件:发酵培养基为TB培养基,接种量为4%,诱导初始菌密度为0.7(OD600)I,PTG浓度为0.2 mmol/L,在20℃下诱导20 h。在最佳条件下,酰基转移酶LovD的表达水平为100 mg/L,辛伐他汀的产量为1.2 g/L。
Simvastatin was synthesized from Monacolin J using acyhransferase LovD. The objective of this study was to investigate the expression of acyltransferase LovD, and optimize its medium and fermenting conditions. The level of expression was visualized by sodium dodecyl sulfate-polyacryl-amide gel electro- phoresis. To quantify the active amount of LovD expressed under different growth conditions and in different media, an activity assay was used. The optimum fermentation conditions of acyltransferase Lord were determined as follows：fermentation medium was Terrific Broth; inoculation volume was 4% ; cell density （ OD600 ） before induction was 0.7 ; concentration of IPTG was 0. 2 mmol/L; inoculation tempreture was 20 ℃, and total time of inoculation was 20 h. Under the conditions, the expression level of acyhransferase LovD was 100 mg/L and the yield of simvastatin was 1.2 g/L.