文献类型：期刊文献

中文题名：重组大肠杆菌发酵生产谷胱甘肽的氨基酸添加策略优化

英文题名：Optimizing the Addition of Amino Acids in the Production of Glutathione by Recombinant Escherichia coli

作者：王德正[1];吴辉[1];李志敏[1];叶勤[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：2015

卷号：31

期号：9

起止页码：197

中文期刊名：生物技术通报

外文期刊名：Biotechnology Bulletin

收录：CSTPCD;;北大核心:【北大核心2014】；CSCD:【CSCD_E2015_2016】；

基金：国家自然科学青年基金项目(21406065);上海市科委产学研医合作项目(13DZ1930200);生物反应器工程国家重点实验室开放课题资助项目(2060204)

语种：中文

中文关键词：谷胱甘肽;氨基酸;半胱氨酸;重组大肠杆菌

外文关键词：glutathione; amino acid; cysteine; recombinant Escherichia coli;

摘要：对表达双功能谷胱甘肽合成酶的重组大肠杆菌发酵生产谷胱甘肽(Glutathione,GSH)进行氨基酸添加策略优化,结果表明:基本培养基中未添加氨基酸时GSH产量为0.81 g/L;诱导2 h后添加17 mmol/L半胱氨酸GSH产量为1.16 g/L,比不加氨基酸提高43%;添加17 mmol/L的3种前体氨基酸,GSH产量达到3.86 g/L,比只添加半胱氨酸提高2.33倍;进一步提高3种氨基酸添加量至25 mmol/L,GSH产量可达4.64 g/L,比不添加氨基酸提高4.73倍,总生产强度高达317.8 mg/(L·h),半胱氨酸转化为谷胱甘肽达到0.60 mol/mol;考察氨基酸添加模式发现一次性添加25 mmol/L氨基酸较恒速流加模式生产速率提高了29.8%。后续在50 L罐放大生产GSH,产量为4.31 g/L,总生产强度达到310.1 mg/(L·h),为工业化放大生产GSH奠定了基础。
The condition of amino acids was optimized in the production of glutathione（GSH）by recombinant Escherichia coli expressing bifunctional glutathione synthetase. Only 0.81 g/L of GSH was produced without adding any amino acid, whereas 1.16 g/L of GSH was formed when 17 mmol/L of cysteine was fed into the medium after 2 h induction, increasing 43%compared to no amino acid added. With the feeding 17 mmol/L of 3 precusor amino acids, the production was improved significantly and achieved at 3.86 g/L, which was 2.33 times of single cysteine addition. By increasing the amount of 3 amino acids to 25 mmol/L, GSH was 4.64 g/L, 4.73 times as no amino acid added, and the overall productivity was 317.8 mg/（L·h）, the conversion rate of cysteine to glutathione was 0.60 mol/mol. While concerning the addition mode, the production rate of GSH was 3.09 g/（L·h）when 25 mmol/L of amino acids was once added to the fermenter, only 2.38 g/（L·h） when 25 mmol/L of amino acids added at constant speed and continuously, so the productivity increased 29.8%by adding once. The process was successfully scaled up in the 50 L fermenter and 4.31 g/L of GSH was produced with an overall productivity of 310.1 mg/（L·h）, which laid a foundation for the industrial production of GSH.