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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.
Afiliação
  • Mishra S; Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706,  USA.
  • Perkovich PM; Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706,  USA.
  • Mitchell WP; LanzaTech, Skokie, IL 60077, USA.
  • Venkataraman M; Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706,  USA.
  • Pfleger BF; Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706,  USA.
Article em En | MEDLINE | ID: mdl-38366943
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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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cupriavidus necator Idioma: En Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cupriavidus necator Idioma: En Ano de publicação: 2024 Tipo de documento: Article