Zhuang Wenjin, Liu Hanyu, Zhang Ying, He Junyao, Wang Pu
Key Laboratory of Green Pharmaceutical Technologies and Related Equipment of Ministry of Education, College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310014, China.
Zhejiang Pharmaceutical College, Ningbo, 315100, China.
AMB Express. 2021 Aug 19;11(1):118. doi: 10.1186/s13568-021-01278-6.
(R)-1-[3-(Trifluoromethyl)phenyl]ethanol ((R)-MTF-PEL) is an important chiral building block for the synthesis of a neuroprotective compound, (R)-3-(1-(3-(trifluoromethyl)phenyl)ethoxy)azetidine-1-carboxamide. In this work, an effective whole-cell-catalyzed biotransformation was developed to produce (R)-MTF-PEL, and its productivity was increased by medium engineering strategy. The recombinant E. coli BL21(DE3)-pET28a(+)-LXCAR-S154Y variant affording carbonyl reductase was adopted for the reduction of 3'-(trifluoromethyl)acetophenone to (R)-MTF-PEL with enantiomeric excess (ee) > 99.9%. The addition of 0.6% Tween-20 (w/v) boosted the bioreduction, because the substrate concentration was increased by 4.0-fold than that in the neat buffer solution. The biocatalytic efficiency was further enhanced by introducing choline chloride: lysine (ChCl:Lys, molar ratio of 1:1) in the reaction medium, because the product yield reached 91.5% under 200 mM substrate concentration in the established Tween-20/ChCl:Lys-containing system, which is the highest ever reported for (R)-MTF-PEL production. The optimal reduction conditions were as follows: 4% (w/v) ChCl:Lys, 12.6 g (DCW)/L recombinant E. coli cells, pH 7.0, 30 ℃ and 200 rpm, reaction for 18 h. The combined strategy of surfactant and NADES has great potential in the biocatalytic process and the synthesis of chiral alcohols.
(R)-1-[3-(三氟甲基)苯基]乙醇((R)-MTF-PEL)是合成神经保护化合物(R)-3-(1-(3-(三氟甲基)苯基)乙氧基)氮杂环丁烷-1-甲酰胺的重要手性结构单元。在本研究中,开发了一种有效的全细胞催化生物转化方法来生产(R)-MTF-PEL,并通过培养基工程策略提高了其生产效率。采用提供羰基还原酶的重组大肠杆菌BL21(DE3)-pET28a(+)-LXCAR-S154Y变体将3'-(三氟甲基)苯乙酮还原为对映体过量(ee)>99.9%的(R)-MTF-PEL。添加0.6%吐温-20(w/v)促进了生物还原,因为底物浓度比纯缓冲溶液中提高了4.0倍。通过在反应介质中引入氯化胆碱:赖氨酸(ChCl:Lys,摩尔比1:1)进一步提高了生物催化效率,因为在建立的含吐温-20/ChCl:Lys的体系中,在200 mM底物浓度下产物收率达到91.5%,这是(R)-MTF-PEL生产中报道的最高收率。最佳还原条件如下:4%(w/v)ChCl:Lys,12.6 g(DCW)/L重组大肠杆菌细胞,pH 7.0,30℃和200 rpm,反应18 h。表面活性剂和天然深共晶溶剂的联合策略在生物催化过程和手性醇的合成中具有巨大潜力。