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Enhanced production of l-methionine in engineered Escherichia coli with efficient supply of one carbon unit

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Abstract

Objective

l-methionine is an important sulfur-containing amino acid essential for humans and animals. Its biosynthesis pathway is complex and highly regulated. This study aims to explore the bottleneck limiting the improvement of l-methionine productivity and apply efficient strategies to increase l-methionine production in engineered E. coli.

Results

The enzyme O-succinylhomoserine sulfhydrylase involved in thiolation of OSH to form homocysteine was overexpressed in the engineered strain E. coli W3110 IJAHFEBC/PAm, resulting in l-methionine production increased from 2.8 to 3.22 g/L in shake flask cultivation. By exogenous addition of l-glycine as the precursor of one carbon unit, the titer of l-methionine was increased to 3.68 g/L. The glycine cleavage system was further strengthened for the efficient one carbon unit supply and a l-methionine titer of 3.96 g/L was obtained, which was increased by 42% compared with that of the original strain.

Conclusions

Insufficient supply of one carbon unit was found to be the issue limiting the improvement of l-methionine productivity and its up-regulation significantly promoted the l-methionine production in the engineered E. coli.

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Acknowledgements

This work was supported by National Key Research and Development Program of China (2018YFA0901403) and National Natural Science Foundation of China (31971342 and 31700095).

Supporting Information

Supplementary Table 1—Plasmids and strains used in this study.

Supplementary Table 2—Primers used in this study.

Supplementary Table 3—The nucleotide sequences of codon-optimized genes used in this study.

Supplementary Table 4—Comparison of L-methionine productivity in different engineered strains.

Supplementary Fig. 1—The procedure to construct the recombinant plasmids. (Take the PAmZ plasmid construction as an example).

Supplementary Fig 2—Quantitative RT-PCR results of metC and malY genes in M2/PAm, M2/PAmC and M2/PAmlY, respectively.

Supplementary Fig. 3—Residual amount of extracellular glycine in M2/PAmZ strain after adding different concentrations of L-glycine in the fermentation culture.

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Correspondence to Zhi-Qiang Liu.

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Tang, XL., Du, XY., Chen, LJ. et al. Enhanced production of l-methionine in engineered Escherichia coli with efficient supply of one carbon unit. Biotechnol Lett 42, 429–436 (2020). https://doi.org/10.1007/s10529-019-02786-z

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  • DOI: https://doi.org/10.1007/s10529-019-02786-z

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