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The limited number of well-characterized model bacteria cannot address all the challenges in a circular bioeconomy [1]. Therefore, there is a growing demand for novel platform organisms with enhanced resistance to extreme conditions, versatile metabolic capabilities, and the ability to utilize cost-effective renewable resources while efficiently generating desirable biobased products [2]. Gram-negative (G-) thermophilic bacteria are attractive microbial platforms, promising significant advances in industrial and environmental applications. However, compared to model mesophilic bacteria or some Gram-positive thermophilic counterparts, the genetic engineering toolkits for G- thermophiles are practically non-existent.
This lecture will summarize recent efforts of a local consortium of several laboratories across Masaryk University and Brno University of Technology in developing and applying genetic and systems metabolic engineering tools for Caldimonas thermodepolymerans, a facultative G- thermophile that efficiently converts a spectrum of plant biomass sugars into high quantities of polyhydroxyalkanoates—fully biodegradable substitutes for synthetic plastics—and other valuable bioproducts [3]. Due to its thermophilic nature, polymer production capacity, and environmental resilience, this bacterium is an attractive candidate for next-generation industrial bioprocesses [4].
Our current work includes optimizing the DNA transformation protocol, developing and testing plasmid-based tools for genome editing, including thermophilic CRISPR/Cas technology, identifying innate regulatory sequences, developing a genome-scale metabolic model, and making initial progress in engineering chassis strains with enhanced bioproduction properties and facilitated genetic manipulation. Our work pioneers the biotechnological domestication of C. thermodepolymerans and related G- thermophiles, paving the way for better understanding their extremophilic nature and boosting their bioproduction potential through genetic and metabolic engineering.
References
[1] Silva M, Donati S, Dvořák P. Current Opinion in Biotechnology (2025), 92:103270.
[2] Obruča S, Dvořák P, Sedlaček P, et al. Biotechnology Advances (2022), 58:107906.
[3] Grybchuk-Ieremenko A, Lipovská K, Kouřilová X, Obruča S, Dvořák P. Microbial Biotechnology (2025), 18(2):e70103.
[4] Chen GQ, Jiang XR. Current Opinion in Biotechnology (2018), 50:94.
