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An acetate-independent pathway for isopropanol production via HMG-CoA in Escherichia coli.
- Source :
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Journal of biotechnology [J Biotechnol] 2022 Nov 20; Vol. 359, pp. 29-34. Date of Electronic Publication: 2022 Sep 20. - Publication Year :
- 2022
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Abstract
- Isopropanol has a good potential as a new fuel substitution. In the model biosynthesis pathway of isopropanol synthesis, acetoacetyl-CoA is converted to acetoacetate by acetoacetyl-CoA transferases, which requires an acetate molecule as a substrate. Herein, a novel isopropanol synthesis pathway based on mammalian ketone metabolic pathway was developed. In this pathway, acetoacetyl-CoA is condensed with acetyl-CoA to generate 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) by HMG-CoA synthase, and then catalyzed by HMG-CoA lyase to generate acetoacetate. This process is acetate-independent. Under the same experimental system using glycerol as carbon source, the E. coli strain MG::ISOP1 containing the novel pathway produced 11.7 times more isopropanol than the strain MG::ISOP0 containing the model pathway. The pta-ackA knockout mutant strain MG∆pta-ackA::ISOP1, which reduced the conversion of acetyl-CoA to acetate, further increased the production from 76 mg/L to 360 mg/L. In another strategy, knocking out atoDA to block the acetoacetate degradation pathway in strain MG∆atoDA::ISOP1 increased the production to 680 mg/L. By knocking out both of pta-ackA and atoDA, strain MGΔpta-ackAΔatoDA::ISOP1 produced 964 mg/L of isopropanol, which was 12.7 times that of MG::ISOP1. This study indicated that the novel pathway is competent for isopropanol synthesis, and provides a new perspective for biosynthesis of isopropanol.<br />Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.<br /> (Copyright © 2022 Elsevier B.V. All rights reserved.)
- Subjects :
- Acetoacetates metabolism
Acetyl Coenzyme A metabolism
Coenzyme A-Transferases metabolism
Hydroxymethylglutaryl-CoA Synthase genetics
Hydroxymethylglutaryl-CoA Synthase metabolism
Glycerol metabolism
Acetates metabolism
Carbon metabolism
Escherichia coli genetics
Escherichia coli metabolism
2-Propanol metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1873-4863
- Volume :
- 359
- Database :
- MEDLINE
- Journal :
- Journal of biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 36150604
- Full Text :
- https://doi.org/10.1016/j.jbiotec.2022.09.011