Development of a multi-enzymatic desymmetrization and its application for the biosynthesis of l-norvaline from dl-norvaline |
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| Affiliation: | 1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu Province 214122, PR China;2. Jiangnan University (Rugao), Food Biotechnology Research Institute, Rugao, Jiangsu Province 226500, PR China;3. Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, United States;1. School of Chemistry, University of Manchester, Manchester Institute of Biotechnology (MIB),131 Princess Street, Manchester, M1 7DN, United Kingdom;2. Facultad de Quimica, Universidad de la República, Montevideo, Uruguay;3. Dr. Reddy’s Laboratories, Chirotech Technology Centre, 410 Cambridge Science Park, Cambridge, CB4 0PE, United Kingdom;1. School of Pharmaceutical Sciences (Shenzhen), Sun Yat-sen University, Shenzhen, 518107, China;2. School of Food Science and Engineering, South China University of Technology, Guangzhou, 510640, China;1. Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China;2. School of Pharmaceutical Engineering, Zhejiang Pharmaceutical College, Ningbo 315100, PR China |
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| Abstract: | Perindopril is an effective antihypertensive drug in strong demand used to treat hypertension. l-norvaline is a vital intermediate of Perindopril production mainly produced by chemical synthesis with low purity. We developed an environmentally friendly method to produce l-norvaline with high purity based on a desymmetrization process. d-Norvaline was oxidized to the corresponding keto acid by d-amino acid oxidase from the substrate dl-norvaline. Asymmetric hydrogenation of the keto acid to l-norvaline was carried out by leucine dehydrogenase with concomitant oxidation of NADH to NAD+. A NADH regeneration system was introduced by overexpressing a formate dehydrogenase. The unwanted H2O2 by-product generated during d-norvaline oxidation was removed by adding catalase. A total of 54.09 g/L of l-norvaline was achieved, with an enantiomeric excess over 99% under optimal conditions, with a 96.7% conversion rate. Our desymmetrization method provides an environmental friendly strategy for the production of enantiomerically pure l-norvaline in the pharmaceutical industry. |
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| Keywords: | Desymmetrization Leucine dehydrogenase NADH regeneration |
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