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[1]刘巧利 杨套伟 周俊平 徐美娟 张 显 饶志明**.酶法高效转化苯乙酮酸合成L-苯甘氨酸*[J].应用与环境生物学报,2019,25(02):1-10.[doi:10.19675/j.cnki.1006-687x.2018.07009]
 LIU Qiaoli,YANG Taowei,ZHOU Junping,et al.Efficient enzymatic synthesis of L-phenylglycine from benzoylformic acid *[J].Chinese Journal of Applied & Environmental Biology,2019,25(02):1-10.[doi:10.19675/j.cnki.1006-687x.2018.07009]
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酶法高效转化苯乙酮酸合成L-苯甘氨酸*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年02期
页码:
1-10
栏目:
研究论文
出版日期:
2019-04-25

文章信息/Info

Title:
Efficient enzymatic synthesis of L-phenylglycine from benzoylformic acid *
文章编号:
201807009
作者:
刘巧利 杨套伟 周俊平 徐美娟 张 显 饶志明**
江南大学生物工程学院工业生物技术教育部重点实验室 无锡 214122
Author(s):
LIU Qiaoli YANG Taowei ZHOU Junping XU Meijuan ZHANG Xian & RAO Zhiming**
School of Biotechnology, Jiangnan University, Ministry of Education Key Laboratory of Industrial Biotechnology, Wuxi 214122, China
关键词:
L-苯甘氨酸苯乙酮酸亮氨酸脱氢酶甲酸脱氢酶辅酶再生体系
Keywords:
L-phenylglycine benzoylformic acid leucine dehydrogenase formate dehydrogenase cofactor regeneration system
DOI:
10.19675/j.cnki.1006-687x.2018.07009
摘要:
L-苯甘氨酸是合成多种抗生素和抗癌药物的重要中间体,目前,其主要通过化学法合成。本研究利用蜡样芽孢杆菌来源的亮氨酸脱氢酶(LeuDH)催化苯乙酮酸的还原氨基化合成L-苯甘氨酸,并偶联甲酸脱氢酶(FDH)进行辅酶再生,建立了一种新型的苯甘氨酸生物合成方法。结果表明,该辅酶再生体系可有效地用于L-苯甘氨酸的合成,且没有副产物残留,辅底物甲酸铵还可提供还原氨基化所需NH4+,随后对酶转化条件进行优化,最适转化条件为:苯乙酮酸60 g/L,甲酸铵50.4 g/L,LeuDH 4 U/mL,FDH 2 U/mL,NAD+ 浓度为0.14 g/L,pH 8.0,以及30 。在最优条件下,1 L的转化体系中,转化反应5 h,苯乙酮酸转化率达到99%,L-苯甘氨酸产量60.2 g/L,ee值>99%。本研究为L-苯甘氨酸的工业生产提供了一种更加简单、高效、经济的生物合成途径。(图8 表4 参22 )
Abstract:
L-phenylglycine, an important intermediate in the synthesis of various antibiotics and anticancer drugs, is mainly synthesized through chemical methods now. In this study, a new biological synthesis method of L-phenylglycine was developed, in which Leucine dehydrogenase(LeuDH) from Bacillus cereus was used to catalyze the reductive amination of benzoylformic acid for L-phenylglycine synthesis and formate dehydrogenase (FDH) was used for cofactor regeneration. The results showed that the cofactor regeneration system was effective for the synthesis of L-phenylglycine without any by-products, and the reaction cosubstrate ammonium formate further provided the ammonium ions (NH4+) required for reductive amination. The enzymatic transformation conditions were then optimized for improved conversion efficiency. The optimal conversion conditions obtained were: benzoylformic acid 60 g/L, ammonium formate 50.4 g/L, LeuDH 4 U/mL, FDH 2 U/mL, NAD+ concentration 0.14 g/L, pH 8.0, and 30 °C. Finally, under these optimal conditions, 99% conversion rate was achieved in 5 h with the yield of 60.2 g/L, and ee value >99% at 1 L scale.This work therefore provides a simpler, more efficient and economical biosynthetic method for L-phenylglycine production.

备注/Memo

备注/Memo:
收稿日期: 2018-07-07 接受日期 Accepted: 2018-07-26
*江苏省杰出青年科学基金(BK20150002)、国家自然科学基金(21778024)、中国博士后科学基金(2017M620189)、国家轻工技术与工程一流学科自主课题资助项目(LITE2018-06)、中央高校基本科研业务费专项资金(JUSRP51708A)和江苏高校优势学科建设工程资助项目资助?
**通讯作者(E-mail: raozhm@jiangnan.edu.cn)
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更新日期/Last Update: 2018-08-30