Unlocking Bacterial Cellulose Functionalisation: Comparative Genetic and Co-Culture Strategies in Komagataeibacter

Abstract Bacterial cellulose (BC) is produced by diverse bacterial species, including members of the genus Komagataeibacter , and has emerged as a potential sustainable alternative to conventional materials such as plastics and leather. Its production by bacteria is rapid and easily scalable, and via genetic modifications or culturing the bacteria with other microbes, it is possible to rationally enhance the BC that is produced, for example by adding functional components. Here, we systematically evaluated BC functionalization strategies, including genetic engineering and co-culturing with other engineered microbes across the most widely used Komagataeibacter species: K. rhaeticus , K. xylinus , K. medellinensis and K. sucrofermentans . We established that all tested strains are amenable to DNA transformation and capable of expressing heterologous genes from identical genetic constructs. However, we identified species-specific differences in heterologous gene expression, cellulose production in varying environmental conditions and in the abilities of the strains to co-culture with Escherichia coli . Additionally, we demonstrated that all these cellulose-producing bacteria can establish functional symbiotic co-cultures with yeast. While inter-species variations in heterologous gene expression and co-culture dynamics are evident, BC modification through genetic engineering and co-culturing strategies remains achievable across all tested Komagataeibacter species. Our work provides a comparative framework to guide researchers in selecting optimal species based on their specific application requirements.