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Metabolic engineering v.44, 2017년, pp.143 - 149   SCIE
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Cofactor self-sufficient whole-cell biocatalysts for the production of 2-phenylethanol

Wang, Pengchao (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 ) ; Yang, Xinwei (National Engineering Research Center of Industrial Microbiology and Fermentation Technology ) ; Lin, Baixue (College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350108, China ) ; Huang, Jianzhong (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 ) ; Tao, Yong (National Engineering Research Center of Industrial Microbiology and Fermentation Technology ) ;
  • 초록  

    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. Highlights A cofactor self-sufficient system was designed by a “bridge” between the amino acid and its corresponding alcohol. By borrowing hydrogen from glutamate dehydrogenase reaction for reduction of the final step of 2-PE synthesis, a cofactor self-sufficient system was developed. Zeolite was utilized to remove the only waste ammonia and in situ product removal technology was applied to alleviate inhibition and toxicity of product. Thus a green and efficient whole-cell biocatalysis process was developed. This study offers a new method to resolve the cofactor/redox imbalance problem.


  • 주제어

    Whole-cell biocatalyst .   Cofactor .   Redox .   2-oxoglutarate .   Self-sufficient .   2-phenylethanol.  

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