Abstract
Phosphinothricin (PPT) is one of the most prevalently using herbicides. The commercial phosphinothricin products are generally in the form of a racemic mixture, of which only the l-phosphinothricin (L-PPT) gives herbicidal function. Synthesis of optically pure L-PPT by deracemization of D/L-PPT is a promising way to cut down the environmental burden and manufacturing cost. To convert D/L-PPT to L-PPT, we expressed the catalytic enzymes by genomic integration in E. coli. The whole production was implemented in two steps in one pot using four catalytic enzymes, namely d-amino acid oxidase, catalase, glutamate dehydrogenase, and glucose dehydrogenase. Finally, after a series of process optimization, the results showed that with our system the overall L-PPT yield reached 86%. Our study demonstrated a new strategy for L-PPT synthesis, based on enzymes from chromosomal integrated expression, which does not depend on antibiotic selection, and shows a high potential for future industrial application.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alpdagtas S, Yucel S, Kapkac HA, Liu SQ, Binay B (2018) Discovery of an acidic, thermostable and highly NADP(+) dependent formate dehydrogenase from Lactobacillus buchneri NRRL B-30929. Biotechnol Lett 40(7):1135–1147. https://doi.org/10.1007/s10529-018-2568-6
Brunhuber NMW, Blanchard JS (1994) The biochemistry and enzymology of amino-acid dehydrogenases. Crit Rev Biochem Molec Biol 29(6):415–467. https://doi.org/10.3109/10409239409083486
Cao CH, Gong H, Dong Y, Li JM, Cheng F, Xue YP, Zheng YG (2021) Enzyme cascade for biocatalytic deracemization of D, L-phosphinothricin. J Biotechnol 325:372–379. https://doi.org/10.1016/j.jbiotec.2020.09.024
Cheng F, Li H, Zhang K, Li QH, Xie D, Xue YP, Zheng YG (2020) Tuning amino acid dehydrogenases with featured sequences for L-phosphinothricin synthesis by reductive amination. J Biotechnol 312:35–43. https://doi.org/10.1016/j.jbiotec.2020.03.001
Chung ME, Yeh IH, Sung LY, Wu MY, Chao YP, Ng IS, Hu YC (2017) Enhanced integration of large DNA Iinto E. coli chromosome by CRISPR/Cas9. Biotechnol Bioeng 114(1):172–183. https://doi.org/10.1002/bit.26056
Englaender JA, Jones JA, Cress BF, Kuhlman TE, Linhardt RJ, Koffas MAG (2017) Effect of genomic integration location on heterologous protein expression and metabolic engineering in E. coli. ACS Synth Biol 6(4):710–720. https://doi.org/10.1021/acssynbio.6b00350
Hernandez K, Berenguer-Murcia A, Rodrigues RC, Fernandez-Lafuente R (2012) Hydrogen peroxide in biocatalysis. A dangerous liaison. Curr Org Chem 16(22):2652–2672. https://doi.org/10.2174/138527212804004526
Horsman GP, Zechel DL (2017) Phosphonate biochemistry. Chem Rev 117(8):5704–5783. https://doi.org/10.1021/acs.chemrev.6b00536
Jia XB, Chen JC, Lin CQ, Lin XJ (2016) Cloning, expression, and characterization of a novel thermophilic monofunctional catalase from Geobacillus sp CHB1. Biomed Res Int 2016:8. https://doi.org/10.1155/2016/7535604
Jia DX, Liu ZJ, Xu HP, Li JL, Li JJ, Jin LQ, Cheng F, Liu ZQ, Xue YP, Zheng YG (2019) Asymmetric synthesis of L-phosphinothricin using thermostable alpha-transaminase mined from Citrobacter koseri. J Biotechnol 302:10–17. https://doi.org/10.1016/j.jbiotec.2019.06.008
Jia DX, Xu HP, Sun CY, Peng C, Li JL, Jin LQ, Cheng F, Liu ZQ, Xue YP, Zheng YG (2020) Covalent immobilization of recombinant Citrobacter koseri transaminase onto epoxy resins for consecutive asymmetric synthesis of L-phosphinothricin. Bioprocess Biosyst Eng 43(9):1599–1607. https://doi.org/10.1007/s00449-020-02351-3
Jiang Y, Chen B, Duan CL, Sun BB, Yang JJ, Yang S (2015) Multigene editing in the Escherichia coli genome via the CRISPR-Cas9 system. Appl Environ Microbiol 81(7):2506–2514. https://doi.org/10.1128/aem.04023-14
Jin LQ, Peng F, Liu HL, Cheng F, Jia DX, Xu JM, Liu ZQ, Xue YP, Zheng YG (2019) Asymmetric biosynthesis of L-phosphinothricin by a novel transaminase from Pseudomonas fluorescens ZJB09-108. Process Biochem 85:60–67. https://doi.org/10.1016/j.procbio.2019.07.010
Kang XM, Cai X, Liu ZQ, Zheng YG (2019) Identification and characterization of an amidase from Leclercia adecarboxylata for efficient biosynthesis of L-phosphinothricin. Bioresour Technol 289:8. https://doi.org/10.1016/j.biortech.2019.121658
Kwon YD, Kim S, Lee SY, Kim P (2011) Long-term continuous adaptation of Escherichia coli to high succinate stress and transcriptome analysis of the tolerant strain. J Biosci Bioeng 111(1):26–30. https://doi.org/10.1016/j.jbiosc.2010.08.007
Leuchs S, Greiner L (2011) Alcohol dehydrogenase from Lactobacillus brevis: a versatile robust catalyst for Enantioselective Transformations. Chem Biochem Eng Q 25(2):267–281. https://doi.org/10.3103/S036152191103013X
Liu ZQ, Ye JJ, Shen ZY, Hong HB, Yan JB, Lin Y, Chen ZX, Zheng YG, Shen YC (2015) Upscale production of ethyl (S)-4-chloro-3-hydroxybutanoate by using carbonyl reductase coupled with glucose dehydrogenase in aqueous-organic solvent system. Appl Microbiol Biotechnol 99(5):2119–2129. https://doi.org/10.1007/s00253-014-6245-y
Marangoni AG (2003) Enzyme kinetics: a modern approach. Wiley, New York
Nakashima N, Akita H, Hoshino T (2014) Establishment of a novel gene expression method, BICES (biomass-inducible chromosome-based expression system), and its application to the production of 2,3-butanediol and acetoin. Metab Eng 25:204–214. https://doi.org/10.1016/j.ymben.2014.07.011
Sabri S, Steen JA, Bongers M, Nielsen LK, Vickers CE (2013) Knock-in/Knock-out (KIKO) vectors for rapid integration of large DNA sequences, including whole metabolic pathways, onto the Escherichia coli chromosome at well-characterised loci. Microb Cell Fact. https://doi.org/10.1186/1475-2859-12-60
Tas H, Nguyen CT, Patel R, Kim NH, Kuhlman TE (2015) An integrated system for precise genome modification in Escherichia coli. PLOS ONE 10(9):19. https://doi.org/10.1371/journal.pone.0136963
Taylor KB (2002) Enzyme kinetics and mechanisms. Academic Press, Berlin
Trost EM, Fischer L (2002) Minimization of by-product formation during D-amino acid oxidase catalyzed racemate resolution Of D/L-amino acids. J Mol Catal B-Enzym 19:189–195. https://doi.org/10.1016/s1381-1177(02)00166-2
Wang JL, Niyompanich S, Tai YS, Wang JY, Bai WQ, Mahida P, Gao T, Zhang KC (2016) Engineering of a highly efficient escherichia coli strain for mevalonate fermentation through chromosomal integration. Appl Environ Microbiol 82(24):7176–7184. https://doi.org/10.1128/aem.02178-16
Xu JM, Zhang K, Cao HT, Li H, Cheng F, Cao CH, Xue YP, Zheng YG (2021) Development of a biocatalytic cascade for synthesis of 2-oxo-4-(hydroxymethylphosphinyl) butyric acid in one pot. Biocatal Biotransform 39(3):190–197. https://doi.org/10.1080/10242422.2020.1797697
Xue YP, Cao CH, Zheng YG (2018) Enzymatic asymmetric synthesis of chiral amino acids. Chem Soc Rev 47(4):1516–1561. https://doi.org/10.1039/c7cs00253j
Yin XJ, Wu JP, Yang LR (2018) Efficient reductive amination process for enantioselective synthesis of L-phosphinothricin applying engineered glutamate dehydrogenase. Appl Microbiol Biotechnol 102(10):4425–4433. https://doi.org/10.1007/s00253-018-8910-z
Yu HL, Li T, Chen FF, Luo XJ, Li AT, Yang C, Zheng GW, Xu JH (2018) Bioamination of alkane with ammonium by an artificially designed multienzyme cascade. Metab Eng 47:184–189. https://doi.org/10.1016/j.ymben.2018.02.009
Zhang RZ, Zhang BT, Xu Y, Li YH, Li M, Liang HB, Xiao R (2013) Efficicent (R)-phenylethanol production with enantioselectivity-alerted (S)-carbonyl reductase II and NADPH regeneration. PLOS ONE 8(12):10. https://doi.org/10.1371/journal.pone.0083586
Zhang YF, Wang Q, Hess H (2017) Increasing enzyme cascade throughput by pH-engineering the microenvironment of individual enzymes. ACS Catal 7(3):2047–2051. https://doi.org/10.1021/acscatal.6b03431
Zhao DD, Yuan SL, Xiong B, Sun HN, Ye LJ, Li J, Zhang XL, Bi CH (2016) Development of a fast and easy method for Escherichia coli genome editing with CRISPR/Cas9. Microb Cell Fact. https://doi.org/10.1186/s12934-016-0605-5
Acknowledgements
This work was supported by the National Natural Science Foundation of China (No. 21978268).
Author information
Authors and Affiliations
Contributions
Y-KC created the idea and wrote the first draft. HG implemented the laboratory work. Y-PX and Y-GZ provided financial support and finalized the draft.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest in this publication.
Rights and permissions
About this article
Cite this article
Cen, YK., Gong, H., Xue, YP. et al. Biosynthesis of l-phosphinothricin with enzymes from chromosomal integrated expression in E. coli. 3 Biotech 11, 477 (2021). https://doi.org/10.1007/s13205-021-03037-7
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s13205-021-03037-7