Abstract
O-Acetyl-L-homoserine (OAH) is a potentially important platform metabolic intermediate for the production of homoserine lactone, methionine, 1,4-butanediol and 1,3-propanediol which have giant market value. Currently, multiple strategies have been adopted to explore sustainable production of OAH. However, the production of OAH by consuming cheap bio-based feedstocks with Escherichia coli as the chassis is still in its infancy. Construction of high yield OAH-producing strains is of great significance in industry. In this study, we introduced an exogenous metA from Bacillus cereus (metXbc) and engineered an OAH-producing strain by combinatorial metabolic engineering. Initially, exogenous metXs/metA were screened and used to reconstruct an initial biosynthesis pathway of OAH in E. coli. Subsequently, the disruption of degradation and competitive pathways combined with optimal expression of metXbc were carried out, accumulating 5.47 g/L OAH. Meanwhile, the homoserine pool was enriched by overexpressing metL with producing 7.42 g/L OAH. Lastly, the carbon flux of central carbon metabolism was redistributed to balance the metabolic flux of homoserine and acetyl coenzyme A (acetyl-CoA) in OAH biosynthesis with accumulating 8.29 g/L OAH. The engineered strain produced 24.33 g/L OAH with a yield of 0.23 g/g glucose in fed-batch fermentation. By these strategies, the key nodes for OAH synthesis were clarified and corresponding strategies were proposed. This study would lay a foundation for OAH bioproduction.
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The data supporting this study's findings are available at the request of the corresponding author.
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Acknowledgements
We are grateful to Dr. Pil Kim from Department of Biotechnology, Catholic University for providing strain E. coli W3110 (DE3). We are also grateful to Dr. Sheng Yang from Institute of Plant Physiology and Ecology (Chinese Academy of Science, Shanghai) for providing CRISPR-Cas9 plasmids.
Funding
This research 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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YGZ and ZQL conceived of the study. BL wrote the manuscript and LGH, ZQL revised the manuscript. BL, YFY, YYC, XJZ performed the experiments and contributed to data analysis. All authors reviewed and approved the final manuscript.
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Li, B., Huang, LG., Yang, YF. et al. Metabolic engineering and pathway construction for O-acetyl-L-homoserine production in Escherichia coli. 3 Biotech 13, 173 (2023). https://doi.org/10.1007/s13205-023-03564-5
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DOI: https://doi.org/10.1007/s13205-023-03564-5