Dual β-oxidation pathway and transcription factor engineering for methyl ketones production in Saccharomyces cerevisiae
Journal article, 2022

Methyl ketones (MK) are highly valuable fatty acid derivatives with broad applications. Microbes based biosynthesis represents an alternative route for production of these usually fossil based chemicals. In this study, we reported metabolic engineering of Saccharomyces cerevisiae to produce MK, including 2-nonanone, 2-undecanone, 2-tridecanone and 2-pentadecanone. Besides enhancing inherent peroxisomal fatty acids β-oxidation cycle, a novel heterologous cytosolic fatty acids β-oxidation pathway was constructed, and this resulted in an increased production of MK by 2-fold. To increase carbon fluxes to methyl ketones, the supply of precursors was enhanced by engineering lipid metabolism, including improving the intracellular biosynthesis of acyl-CoAs, weakening the consumption of acyl-CoAs for lipids storage, and reinforcing activation of free fatty acids to acyl-CoAs. Hereby the titer of MK was improved by 7-fold, reaching 143.72 mg/L. Finally, transcription factor engineering was employed to increase the biosynthesis of methyl ketones and it was found that overexpression of ADR1 can mimic the oleate activated biogenesis and proliferation of peroxisomes, which resulted in a further increased production of MK by 28%. With these modifications and optimization, up to 845 mg/L total MK were produced from glucose in fed-batch fermentation, which is the highest titer of methyl ketones reported produced by fungi.

Cytosolic β-oxidation

Methyl ketones

Microbial production

Transcription factor engineering

Saccharomyces cerevisiae

Author

Ge Zhang

Chinese Academy of Sciences

National Technology Innovation Center of Synthetic Biology

Chao Zhang

Beijing University of Chemical Technology

Zheng Wang

Beijing University of Chemical Technology

Qinhong Wang

Chinese Academy of Sciences

National Technology Innovation Center of Synthetic Biology

Jens B Nielsen

Beijing University of Chemical Technology

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Zongijie Dai

National Technology Innovation Center of Synthetic Biology

Chinese Academy of Sciences

Metabolic Engineering

1096-7176 (ISSN) 1096-7184 (eISSN)

Vol. 73 225-234

Subject Categories

Chemical Process Engineering

Biocatalysis and Enzyme Technology

Organic Chemistry

DOI

10.1016/j.ymben.2022.08.004

PubMed

35987431

More information

Latest update

9/1/2022 1