重组大肠杆菌发酵生产谷胱甘肽的氨基酸添加策略优化

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.028

生物技术通报 ›› 2015, Vol. 31 ›› Issue (9): 197-203.doi: 10.13560/j.cnki.biotech.bull.1985.2015.09.028

重组大肠杆菌发酵生产谷胱甘肽的氨基酸添加策略优化

王德正, 吴辉, 李志敏, 叶勤

(华东理工大学生物反应器工程国家重点实验室，上海 200237)

收稿日期:2015-01-14 出版日期:2015-09-15 发布日期:2015-09-16
作者简介:王德正，男，硕士，研究方向：发酵工程；E-mail：wdzfromchina@163.com
基金资助:
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)

Optimizing the Addition of Amino Acids in the Production of Glutathione by Recombinant Escherichia coli

Wang Dezheng, Wu Hui, Li Zhimin, Ye Qin

(State Key Laboratory of Bioreactor Engineering，East China University of Science and Technology，Shanghai 200237)

Received:2015-01-14 Published:2015-09-15 Online:2015-09-16

摘要/Abstract

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。

关键词: 谷胱甘肽, 氨基酸, 半胱氨酸, 重组大肠杆菌

Abstract: 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.

Key words: glutathione, amino acid, cysteine, recombinant Escherichia coli

王德正, 吴辉, 李志敏, 叶勤. 重组大肠杆菌发酵生产谷胱甘肽的氨基酸添加策略优化[J]. 生物技术通报, 2015, 31(9): 197-203.

Wang Dezheng, Wu Hui, Li Zhimin, Ye Qin. Optimizing the Addition of Amino Acids in the Production of Glutathione by Recombinant Escherichia coli[J]. Biotechnology Bulletin, 2015, 31(9): 197-203.

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重组大肠杆菌发酵生产谷胱甘肽的氨基酸添加策略优化

Optimizing the Addition of Amino Acids in the Production of Glutathione by Recombinant Escherichia coli

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