The engineering toolbox of Parageobacillus thermoglucosidasius.

The thermophilic bacterium Parageobacillus thermoglucosidasius is considered a promising host for industrial microbial production. However, its genetic engineering toolbox is still under development. The focus of this review is to provide an organised summary of the currently available resources to facilitate the use of this organism. The article offers a comprehensive overview of regulatory parts used for the construction of genetic circuits and plasmids, including promoters, ribosome binding sites, terminators, antibiotic resistance genes, reporter genes, and other genetic elements of interest. Additionally, it examines the developments in P. thermoglucosidasius vector designs over the years. Here, vectors were categorised either as replicative vectors intended for gene expression, or integrative vectors intended for genomic engineering. The functionality of each vector was described, and their contributions to the progress of molecular tools available for P. thermoglucosidasius were evaluated. The review also summarises recent advancements in CRISPR-based systems relevant to this organism. Finally, this review discusses potential improvements that could further contribute to the expanding engineering toolbox of P. thermoglucosidasius, paving the way for more advanced applications. KEY POINTS: • The existing engineering toolbox for P. thermoglucosidasius contributes to the growing interest in using it as a thermophilic production host. • The engineering toolbox for P. thermoglucosidasius has expansion potential in genetic circuit parts and CRISPR-based tools. • Thermophilic hosts like P. thermoglucosidasius are in need of more thermostable reporter genes and thermostable selection markers.