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Metabolic engineering of Escherichia coli for efficient production of l-arginine

Abstract

As an important semi-essential amino acid, l-arginine (l-Arg) has important application prospects in medicine and health care. However, it remains a challenge to efficiently produce l-Arg by Escherichia coli (E. coli). In the present study, we obtained an E. coli A1 with l-Arg accumulation ability, and carried out a series of metabolic engineering on it, and finally obtained an E. coli strain A7 with high l-Arg production ability. First, genome analysis of strain A1 was performed to explore the related genes affecting l-Arg accumulation. We found that gene speC and gene speF played an important role in the accumulation of l-Arg. Second, we used two strategies to solve the feedback inhibition of the l-Arg pathway in E. coli. One was the combination of a mutation of the gene argA and the deletion of the gene argR, and the other was the combination of a heterologous insertion of the gene argJ and the deletion of the gene argR. The combination of exogenous argJ gene insertion and argR gene deletion achieved higher titer accumulation with less impact on strain growth. Finally, we inserted the gene cluster argCJBDF of Corynebacterium glutamicum (C. glutamicum) to enhance the metabolic flux of the l-Arg pathway in E. coli. The final strain obtained 70.1 g/L l-Arg in a 5-L bioreactor, with a yield of 0.326 g/g glucose and a productivity of 1.17 g/(L· h). This was the highest level of l-Arg production by E. coli ever reported. Collectively, our findings provided valuable insights into the possibility of the industrial production of l-Arg by E. coli.

Key points

Genetic background of E. coli A1 genome analysis.

Heterologous argJ substitution of argA mutation promoted excessive accumulation of L-Arg in E. coli A1.

The overexpression of L-Arg synthesis gene cluster argCJBDF of Corynebacterium glutamicum (C. glutamate) promoted the accumulation of L-Arg, and 70.1 g/L L-Arg was finally obtained in fed-batch fermentation.

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Data availability

All data generated or analyzed during this study are included in the published article. Construction of recombinant plasmids and strains; primer pairs used in this study; Analyzing of l-Arg production by shake-flask fermentation.

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Funding

This work was supported by the Green Bio-fabrication Program of China [No. 2021YFC2100900], the Top-Notch Academic Programs Project of Jiangsu Higher Education Institutions, the 111 Project (Grant number 111–2-06).

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Contributions

JX and HW conceived the experiments. HW and WZ designed and performed the experiments and analyzed the data. HW and JX wrote the paper. All authors read and approved the final manuscript.

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Correspondence to Jian-Zhong Xu or Wei-Guo Zhang.

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The authors declare no competing interests.

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Wang, HD., Xu, JZ. & Zhang, WG. Metabolic engineering of Escherichia coli for efficient production of l-arginine. Appl Microbiol Biotechnol 106, 5603–5613 (2022). https://doi.org/10.1007/s00253-022-12109-4

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  • DOI: https://doi.org/10.1007/s00253-022-12109-4

Keywords

  • l-arginine
  • Escherichia coli
  • Metabolic engineering
  • Fermentation
  • argJ