Expanding the synthetic biology toolbox of Cupriavidus necator for establishing fatty acid production

Mishra, Shivangi; Perkovich, Paul M.; Mitchell, Wayne P.; Venkataraman, Maya; Pfleger, Brian F.

doi:10.1093/jimb/kuae008

Title: Expanding the synthetic biology toolbox of Cupriavidus necator for establishing fatty acid production

Journal Article · 15 February 2024 · Journal of Industrial Microbiology and Biotechnology

DOI: https://doi.org/10.1093/jimb/kuae008 · OSTI ID:2322593

Mishra, Shivangi; Perkovich, Paul M.; Mitchell, Wayne P.; Venkataraman, Maya;

Abstract The Gram-negative betaproteobacterium Cupriavidus necator is a chemolithotroph that can convert carbon dioxide into biomass. Cupriavidus necator has been engineered to produce a variety of high-value chemicals in the past. However, there is still a lack of a well-characterized toolbox for gene expression and genome engineering. Development and optimization of biosynthetic pathways in metabolically engineered microorganisms necessitates control of gene expression via functional genetic elements such as promoters, ribosome binding sites (RBSs), and codon optimization. In this work, a set of inducible and constitutive promoters were validated and characterized in C. necator, and a library of RBSs was designed and tested to show a 50-fold range of expression for green fluorescent protein (gfp). The effect of codon optimization on gene expression in C. necator was studied by expressing gfp and mCherry genes with varied codon-adaptation indices and was validated by expressing codon-optimized variants of a C12-specific fatty acid thioesterase to produce dodecanoic acid. We discuss further hurdles that will need to be overcome for C. necator to be widely used for biosynthetic processes.

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Sponsoring Organization:: USDOE

Grant/Contract Number:: AR0001503

OSTI ID:: 2322593

Alternate ID(s):: OSTI ID: 2335326; OSTI ID: 2475923

Journal Information:: Journal of Industrial Microbiology and Biotechnology, Journal Name: Journal of Industrial Microbiology and Biotechnology Vol. 51; ISSN 1367-5435

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: Germany

Language:: English

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