Abstract
Methionine is the only one of the essential amino acids that contain sulfur, widely used as a feed additive in agriculture. In this study, the availability of 5-methyl-tetrahydrofolate was confirmed as the main limitation in the complex multibranched biosynthetic pathway of L-methionine. The cycle of one-carbon units was thoroughly investigated and modified to supply 5-methyl-tetrahydrofolate for L-methionine production, such as enhancing the supply of precursor, expediting the conversion rate of the cycle, introducing exogenous serine hydroxymethyltransferase and increasing pool size of one-carbon units carrier. The final strain MYA/pAmFA-4 was able to produce 20.89 g/L L-methionine by fed-batch fermentation, which was the highest titer reported in the literatures. This study is instructive for other metabolites biosynthesized needing one-carbon units or having a complex multibranched biosynthetic pathway.
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Acknowledgements
We appreciate Dr. Sheng Yang (Institute of Plant Physiology and Ecology, Chinese Academy of Science, Shanghai) providing pTarget and pCas9 for genome edit.
Funding
This work was supported by the National Key Research and Development Project of China (2018YFA0901400) and the National Natural Science Foundation of China (No. 31971342 and 31700095).
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ZYS: Conceptualization, Software, Methodology, Validation, Writing-original draft. YFW: Conceptualization, Software, Methodology, Validation, Writing-original draft. LJW: Writing-review & editing, Conceptualization, Methodology. YW: Methodology and Validation. ZQL: Project administration, Funding acquisition, Supervision, Conceptualization. YGZ: Project administration, Funding acquisition, Supervision, Conceptualization.
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Shen, ZY., Wang, YF., Wang, LJ. et al. Thorough research and modification of one-carbon units cycle for improving L-methionine production in Escherichia coli. 3 Biotech 13, 203 (2023). https://doi.org/10.1007/s13205-023-03625-9
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DOI: https://doi.org/10.1007/s13205-023-03625-9