Abstract
L-arginine (L-Arg) is a semi-essential amino acid with many important physiological functions. However, achieving efficient manufacture of L-Arg on an industrial scale using Escherichia coli (E. coli) remains a major challenge. In previous studies, we constructed a strain of E. coli A7, which had good L-Arg production capacity. In this study, E. coli A7 was further modified, and E. coli A21 with more efficient L-Arg production capacity was obtained. Firstly, we reduced the acetate accumulation of strain A7 by weakening the poxB gene and overexpressing acs gene. Secondly, we improved the L-Arg transport efficiency of strains by overexpressing the lysE gene from Corynebacterium glutamicum (C. glutamicum). Finally, we enhanced the supplies of precursors for the synthesis of L-Arg and optimized the supplies of cofactor NADPH and energy ATP in strain. After fermentation in a 5-L bioreactor, the L-Arg titer of strain A21 was found to be 89.7 g/L. The productivity was 1.495 g/(L·h) and the glucose yield was 0.377 g/g. Our study further narrowed the titer gap between E. coli and C. glutamicum in the synthesis of L-Arg. In all recent studies on the L-Arg production by E. coli, this was the highest titer recorded. In conclusion, our study further promotes the efficient mass synthesis of L-Arg by E. coli.
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Key points
• The acetate accumulation of starting strain A7 was decreased.
• Overexpression of gene lysE of C. glutamicum enhanced L-Arg transport in strain A10.
• Enhance the supplies of precursors for the synthesis of L-Arg and optimize the supplies of cofactor NADPH and energy ATP. Finally, Strain A21 was detected to have an L-Arg titer of 89.7 g/L in a 5-L bioreactor.
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Data availability
All data generated or analyzed during this study are included in the published article. Construction of recombinant plasmid; primer pairs used in this study; analyzing of L-Arg production by shake-flask fermentation.
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Funding
We acknowledge the financial support from the Class I project of independent innovation of agricultural science and technology in Jiangsu Province (grant numbers CX (22) 1012) and the National Key R&D Program “Green Biological Manufacturing” key special project, the “Important Amino Acid Industrial Strain System Transformation and Industrial Demonstration” project (grant numbers 2021YFC2100900).
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H W and J X conceived the experiments. H W and W Z designed and performed the experiments and analyzed the data. H W wrote the paper. All authors read and approved the final manuscript.
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Wang, HD., Xu, JZ. & Zhang, WG. Reduction of acetate synthesis, enhanced arginine export, and supply of precursors, cofactors, and energy for improved synthesis of L-arginine by Escherichia coli. Appl Microbiol Biotechnol 107, 3593–3603 (2023). https://doi.org/10.1007/s00253-023-12532-1
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DOI: https://doi.org/10.1007/s00253-023-12532-1