Expanding the synthetic biology toolbox of Cupriavidus necator for establishing fatty acid production.
J Ind Microbiol Biotechnol
; 512024 Jan 09.
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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1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Cupriavidus necator
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article