Abstract
Objectives
To establish a technique for efficient fatty acid production through enhancement of coenzyme A (CoA) biosynthesis and malonyl-CoA supply by introducing exogenous pantothenate kinase (coaA) and acetyl-CoA carboxylase (acc) in Escherichia coli.
Results
The expression of acc, obtained from Corynebacterium glutamicum, accumulated 2.2-fold more fatty acids in E. coli. The addition of coaA from Pseudomonas putaida or fatty acid synthase (fasA) from C. glutamicum resulted in a 3.1- and 3.6-fold increase in fatty acid synthesis in E. coli cells, which expressed acc and coaA, or acc and fasA, respectively. The transformants, simultaneously possessing all three genes, produced 5.6-fold more fatty acids. The strain possessing acc, coaA, and fasA stored 691 mg/L of fatty acids, primarily as phospholipids, inside the inner membrane after 72-h cultivation. In addition, 19% of the total CoA pool was occupied by malonyl-CoA.
Conclusions
Increased malonyl-CoA significantly contributed to fatty acid production, and the effect was boosted by the expanded total CoA pool. Manipulation of the intracellular CoA species is effective for fatty acid production in E. coli.
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Acknowledgements
This work was supported by a Grant-in-Aid for Scientific Research (C) 16K07655 from The Japan Society for the Promotion of Science. We would like to thank Editage (www. editage.com) for English language editing.
Supporting information
Supplementary Table 1. Oligonucleotide primers used in this work.
Supplementary Table 2. Glucose consumption, pH of media, and pantothenate kinase (CoaA) activities of the E. coli transformants in time-course experiments (See Fig. 3).
Supplementary Fig. 1. Plasmids used in this work. fasA, Corynebacterium glutamicum fatty acid synthase; acpS, C. glutamicum acyl carrier protein synthase; dtsR1, β-subunit of C. glutamicum acetyl-CoA carboxylase; accBC, α-subunit; accE, ε-subunit; coaA, Pseudomonas putida pantothenate kinase.
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Satoh, S., Ozaki, M., Matsumoto, S. et al. Enhancement of fatty acid biosynthesis by exogenous acetyl-CoA carboxylase and pantothenate kinase in Escherichia coli. Biotechnol Lett 42, 2595–2605 (2020). https://doi.org/10.1007/s10529-020-02996-w
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DOI: https://doi.org/10.1007/s10529-020-02996-w