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Covalent immobilization of recombinant Citrobacter koseri transaminase onto epoxy resins for consecutive asymmetric synthesis of L-phosphinothricin

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Abstract

Transaminase responsible for alienating prochiral ketone compound is applicable to asymmetric synthesis of herbicide L-phosphinothricin (L-PPT). In this work, the covalent immobilization of recombinant transaminase from Citrobacter koseri (CkTA) was investigated on different epoxy resins. Using optimum ES-105 support, a higher immobilized activity was obtained via optimizing immobilization process in terms of enzyme loading, coupling time and initial PLP concentration. Crucially, due to blocking unreacted epoxy groups on support surface with amino acids, the reaction temperature of blocked immobilized biocatalyst was enhanced from 37 to 57 °C. Its thermostability at 57 °C was also found to be superior to that of free CkTA. The Km value was shifted from 36.75 mM of free CkTA to 39.87 mM of blocked immobilized biocatalyst, demonstrating that the affinity of enzyme to the substrate has not been apparently altered. Accordingly, the biocatalyst performed the consecutive synthesis of L-PPT for 11 cycles (yields>91%) with retaining more than 91.13% of the initial activity. The seemingly the highest reusability demonstrates this biocatalyst has prospective for reducing the costs of consecutive synthesis of L-PPT with high conversion.

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Acknowledgements

This project was financially supported by the National Defense Science and Technology Innovation Zone Foundation of China, and the Natural Science Foundation of China [Grant numbers 21978268, 31700693].

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Authors and Affiliations

  1. The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, 18 Chaowang road, Hangzhou, 310014, P. R. China

    Dong-Xu Jia, Hai-Peng Xu, Chen-Yi Sun, Chen Peng, Jun-Liang Li, Li-Qun Jin, Feng Cheng, Zhi-Qiang Liu, Ya-Ping Xue & Yu-Guo Zheng

  2. Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 18 Chaowang road, Hangzhou, 310014, P. R. China

    Dong-Xu Jia, Hai-Peng Xu, Chen-Yi Sun, Chen Peng, Jun-Liang Li, Li-Qun Jin, Feng Cheng, Zhi-Qiang Liu, Ya-Ping Xue & Yu-Guo Zheng

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  1. Dong-Xu Jia
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  2. Hai-Peng Xu
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Correspondence to Ya-Ping Xue.

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Jia, DX., Xu, HP., Sun, CY. et al. Covalent immobilization of recombinant Citrobacter koseri transaminase onto epoxy resins for consecutive asymmetric synthesis of L-phosphinothricin. Bioprocess Biosyst Eng 43, 1599–1607 (2020). https://doi.org/10.1007/s00449-020-02351-3

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