Abstract
Acetoin, an important and high-value added bio-based platform chemical, has been widely applied in fields of foods, cosmetics, chemical synthesis, and agriculture. Lactate is a significant intermediate short-chain carboxylate in the anaerobic breakdown of carbohydrates that comprise ~ 18% and ~ 70% in municipal wastewaters and some food processing wastewaters, respectively. In this work, a series of engineered Escherichia coli strains were constructed for efficient production of acetoin from cheaper and abundant lactate through heterogenous co-expression of fusion protein (α-acetolactate synthetase and α-acetolactate decarboxylase), lactate dehydrogenase and NADH oxidase, and blocking acetate synthesis pathways. After optimization of whole-cell bioconversion conditions, the engineered strain BL-11 produced 251.97 mM (22.20 g/L) acetoin with a yield of 0.434 mol/mol in shake flasks. Moreover, a titer of 648.97mM (57.18 g/L) acetoin was obtained in 30 h with a yield of 0.484 mol/mol lactic acid in a 1-L bioreactor. To the best of our knowledge, this is the first report on the production of acetoin from renewable lactate through whole-cell bioconversion with both high titer and yield, which demonstrates the economy and efficiency of acetoin production from lactate.
Key Points • The lactate dehydrogenases from different organisms were expressed, purified, and assayed. • It is the first time that acetoin was produced from lactate by whole-cell biocatalysis. • The highest titer of 57.18 g/L acetoin was obtained with high theoretical yield in a 1-L bioreactor. |
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This work was supported by the National Key Research and Development Program of China (2021YFC2100700) and the National Natural Science Foundation of China (grant NSFC-21576191).
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TC and ZC conceived and designed the project. ZC carried out the experiments. ZC and ZW analyzed the data. MZ, MD, and WD provided some support during the experimental phase. ZC and TC wrote the manuscript with contributions from all authors. All authors read and approved the final manuscript.
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Cui, Z., Zheng, M., Ding, M. et al. Efficient production of acetoin from lactate by engineered Escherichia coli whole-cell biocatalyst. Appl Microbiol Biotechnol 107, 3911–3924 (2023). https://doi.org/10.1007/s00253-023-12560-x
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DOI: https://doi.org/10.1007/s00253-023-12560-x