One-step eantioselective bioresolution for (S)-2-chlorophenylglycine methyl ester catalyzed by the immobilized Protease 6SD on multi-walled carbon nanotubes in a triphasic system

• Published in: Journal of biotechnology 2021-01, Vol.325, p.294-302
• Main Authors: Weng, Chun-Yue, Wang, Dan-Na, Ban, Shan-Yun, Zhai, Qiu-Yao, Hu, Xin-Yi, Cheng, Feng, Wang, Ya-Jun, Zheng, Yu-Guo
• Format: Article
• Language: English
• Subjects: (S)-2-chlorophenylglycine methyl ester; Enantioselective bioresolution; Immobilization; Protease; Triphasic system
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2020.10.007

•A novel triphasic system was constructed for bioresolution of (S)-2-chlorophenylglycine methyl ester ((S)-1).•Optically pure (S)-1 was prepared via one-step direct bioresolution by the triphasic system.•The triphasic system consisted of the immobilized Protease 6SD on NH2-MWCNT, aqueous and MTBE phases. (S)-2-chlorophenylglycine methyl ester ((S)-1) is a key chiral building block of clopidogrel, which is a widely administered antiaggregatory and antithrombotic drug. Herein, Protease 6SD was covalently immobilized on multi-walled carbon nanotubes (MWCNT), and the as-prepared immobilizate P-6SD@NH2-MWCNT was applied in the enantioselective resolution of (R,S)-1 to yield (S)-1. In order to overcome the poor solubility of (R,S)-1 in aqueous solution, a novel triphasic reaction system constituting P-6SD@NH2-MWCNT, aqueous phase and methyl tert-butyl ether (MTBE) as the organic phase was constructed, which simultaneously improved the substrate solubility and the immobilizate recyclability. Under the optimized reaction conditions, P-6SD@NH2-MWCNT catalyzed 10 mM (R,S)-1 for 2 h, yielding optically pure (S)-1 (>99.0 % ees) with 70.74 % conversion of the (R,S)-1. Moreover, P-6SD@NH2-MWCNT can be reused for 15 batches, displaying an exquisite recycling performance. It is for the first time that enantiomerically pure (S)-1 was successfully synthesized by protease-catalyzed one-step resolution.