Abstract
2-Pyrrolidone is widely used in the textile and pharmaceutical industries. Here, we established a 2-pyrrolidone biosynthesis pathway in Corynebacterium glutamicum, by expressing glutamate decarboxylase (Gad) mutant and β-alanine CoA transferase (Act) which activates spontaneous dehydration cyclization of GABA to form 2-pyrrolidone. Also, the 5′ untranslated regions (UTR) strategy was used to increase the expression of protein. Furthermore, considering the importance of acetyl-CoA in the 2-pyrrolidone synthesis pathway, the acetyl-CoA synthetase (acsA) gene was introduced to convert acetate into acetyl-CoA thus achieving the recyclability of the economy. Finally, the fed-batch fermentation of the final strain in a 5 L bioreactor produced 10.5 g/L 2-pyrrolidone within 78 h, which increased by 42.5% by altering the level of gene expression. This is the first time to build the basic chemical 2-pyrrolidone from glucose in one step in C. glutamicum.
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Acknowledgements
This work was supported by the National key Research and Development Program of China, (No. 2021YFC2100900); National Nature Science Foundation of China, (No. 32070035); Key Research and Development Program of Ningxia Hui Autonomous Region, (No. 2019BCH01002); The Science and Technology Project of Xinjiang Production and Construction Corps, (No. 2019AB009); the project (111-2-06).
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XMJ, GH and MZF designed and conducted the experiments. They also modified and polished the manuscript. HJ made some suggestions for the experiments; ZKY and SML was responsible for the planning and execution. RZM was responsible for the design and review of manuscripts and ensuring that the descriptions are accurate and agreed by all authors.
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Xu, M., Gao, H., Ma, Z. et al. Development of a 2-pyrrolidone biosynthetic pathway in Corynebacterium glutamicum by engineering an acetyl-CoA balance route. Amino Acids 54, 1437–1450 (2022). https://doi.org/10.1007/s00726-022-03174-0
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DOI: https://doi.org/10.1007/s00726-022-03174-0