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Expression of l-phosphinothricin synthesis enzymes in Pichia pastoris for synthesis of l-phosphinothricin

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Abstract

With the ban of highly toxic herbicides, such as paraquat and glyphosate, phosphinothricin (PPT) is becoming the most popular broad-spectrum and highly effective herbicide. The current PPT products in the market are usually a racemic mixture with two configurations, the D-type and L-type, of which only the L-PPT has the herbicidal activity. The racemic product is not atom economic, more toxic and may cause soil damage. Asymmetric synthesis of L-PPT has become a research focus in recent years, while biological synthesis methods are preferred for its character of environmental friendly and requiring less reaction steps when being compared to the chemical methods. We have developed a biological synthesis route to produce optically pure L-PPT from D,L-PPT in two steps using 2-carbonyl-4- (hydroxymethyl phosphonyl) butyric acid as the intermediate. In this study, we expressed the glutamate dehydrogenase and glucose dehydrogenase using Pichia pastoris as the first time. After a series of optimization, the total L-PPT yield reached 84%. The developed synthesis system showed a high potential for future industrial application. Compare to the previous plasmid-carrying-E. coli expression system, the established method may avoid antibiotic usage and provided an alternative way for industrial synthesis of optically pure L-PPT.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 21978268).

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Authors and Affiliations

  1. Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People’s Republic of China

    Yu-Ke Cen, Qi Wang, Huo Gong, Ya-Ping Xue & Yu-Guo Zheng

  2. Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou, 310014, People’s Republic of China

    Yu-Ke Cen, Qi Wang, Huo Gong, Ya-Ping Xue & Yu-Guo Zheng

  3. National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou, 310014, People’s Republic of China

    Yu-Ke Cen, Qi Wang, Huo Gong, Ya-Ping Xue & Yu-Guo Zheng

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  1. Yu-Ke Cen
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  2. Qi Wang
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  3. Huo Gong
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  4. Ya-Ping Xue
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  5. Yu-Guo Zheng
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Contributions

Y-KC created the idea and wrote the first draft. QW and HG implemented the laboratory work. QW also helped in manuscript revision. Y-PX and Y-GZ provided financial support and finalized the draft.

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Correspondence to Ya-Ping Xue.

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Cen, YK., Wang, Q., Gong, H. et al. Expression of l-phosphinothricin synthesis enzymes in Pichia pastoris for synthesis of l-phosphinothricin. Biotechnol Lett 44, 561–570 (2022). https://doi.org/10.1007/s10529-022-03239-w

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  • DOI: https://doi.org/10.1007/s10529-022-03239-w

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