Cofactor self-sufficient whole-cell biocatalysts for the production of 2-phenylethanol |
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| Affiliation: | 1. CAS Key Laboratory of Microbial Physiological and Metabolic Engineering and State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;2. College of Life Science, University of Chinese Academy of Sciences, Beijing 100049, China;3. National Engineering Research Center of Industrial Microbiology and Fermentation Technology; College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350108, China;1. Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;2. University of Chinese Academy of Sciences, Beijing 100049, China;3. School of Life Sciences, Taishan Medical University, Taian 271000, Shandong Province, China;1. Institute of Microbiology, Chinese Academy of Sciences, CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Beijing, China;2. College of Life Science, University of Chinese Academy of Sciences, Beijing, China;3. University of Science and Technology of China, Anhui, China |
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| Abstract: | The efficiency of biocatalysis is often affected by an insufficient supply and regeneration of cofactors and redox equivalents. To alleviate this shortcoming, a cofactor self-sufficient system was developed for enhanced production of 2-phenylethanol (2-PE) in E. coli. A “bridge” between the amino acid and its corresponding alcohol was designed in the system using glutamate dehydrogenase. By coupling glutamate dehydrogenase with transaminase and alcohol dehydrogenase, the cosubstrate (2-oxoglutarate) and redox equivalents (NAD(P)H) were regenerated simultaneously, so that no external cofactor or redox source was required. Thus, a cofactor self-sufficient system was developed, which improved the biocatalyst efficiency 3.8-fold. The ammonium generated in this process was removed using zeolite, which further improved the biosynthetic efficiency and resulted in a cleaner system. To the best of our knowledge, this system yielded the highest titer of 2-PE ever obtained in E. coli. Additionally, the wider applicability of this self-sufficient strategy was demonstrated in the production of D-phenyllactic acid. This study thus offers a new method to resolve the cofactor/redox imbalance problem and demonstrates the feasibility of the cofactor self-sufficient strategy for enhanced production of diverse chemicals. |
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| Keywords: | Whole-cell biocatalyst Cofactor Redox 2-oxoglutarate Self-sufficient 2-phenylethanol |
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