重组大肠杆菌双酶一锅法全细胞合成L-苯乳酸研究
Study on One-pot,Two-enzyme biosynthesis of L-phenyllactic by whole cell recombinant E. coli
本研究构建了苯丙氨酸脱氢酶和L-羟基异己酸还原酶的共表达重组大肠杆菌E. coli BL21（DE3, pET28a-pdh-ldh），利用全细胞催化L-苯丙氨酸合成L-苯乳酸，且该反应过程能实现辅酶I NAD+和NADH的自循环。对重组大肠杆菌的诱导条件进行了优化，通过单因素试验和正交试验，最终确定了最佳的诱导条件：OD600=0.6时，添加诱导剂IPTG至终浓度为0.8 mmol/L，在22℃下诱导14.0 h。在最佳的诱导条件下得到的重组大肠杆菌E. coli BL21（DE3, pET28a-pdh-ldh）全细胞催化50.0 mmol/L的L-苯丙氨酸，L-苯乳酸的产物浓度能达到18.2 mmol/L，产率为33.0%，显示了较好的应用前景，也为后续酶的改造奠定了基础。
In this study, recombinant Escherichia coli BL21(DE3, pET28a-pdh-ldh) with phenylalanine dehydrogenase and L-isocaproate reductases was constructed.L-phenyllactic acid was synthesized from L-phenylalanine by whole cell.The reaction process can achieve self-circulation of coenzymes I NAD+ and NADH. This experiment also optimized the induction conditions of recombinant E.coli BL21 (DE3, pET28a-pdh-ldh). In the whole cell biosynthesis process using collected recombinant cells as catalyst, at optimal induction conditions(OD600=0.6, IPTG concentration：0.8mmol/L, 22℃and 14.0h), the recombinant E. coli BL21(DE3, pET28a-pdh-ldh) produced 18.2 mmol/L L-PLA with a conversion of33.0% from 50.0 mmol/L L-phenylalanine,which showed a good application prospect and laid the foundation for the subsequent enzyme modification.
王周平、夏雨、茅菁菁
生物工程学生物化学
L-苯乳酸苯丙氨酸脱氢酶L-羟基异己酸还原酶全细胞转化
L-phenyllactic acidPhenylalanine dehydrogenaseL-isocaproate reductaseswhole cell biosynthesis
王周平,夏雨,茅菁菁.重组大肠杆菌双酶一锅法全细胞合成L-苯乳酸研究[EB/OL].(2019-01-28)[2025-08-11].http://www.paper.edu.cn/releasepaper/content/201901-185.点此复制
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