Bacterial cellulose (BC) is a unique biopolymer synthesised by many bacteria as a critical element of their biofilm matrix. The most known and efficient producers of BC are bacteria from the genus of Komagataeibacter. Bacterial cellulose, with its unique properties, high crystallinity, mechanical strength, and unprecedented ability to hold water, is an object of interest in many industries. Despite the enormous efforts that have been made to develop an effective process, the economic aspect of BC production is still a limiting factor for broadening applications, and new “breaking point” solutions are highly anticipated. In this study, the possibility of using sucrose, lactose, and starch as alternative carbon sources converted to simple sugars directly in the culture medium by microbial glycohydrolases, β-D-fructofuranosidase, β-galactosidase, and glucoamylase in the process of BC synthesis was analysed. The results showed the high potential of the enzyme-assisted fermentation process that, for most used raw carbons sources, was highly efficient, with a yield higher (i.e., lactose 40% more) or comparable to the cultures maintained on standard Hestrin-Schramm media with glucose as a sole carbon source. The X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscope analyses did not reveal any negative influence of enzyme-assisted cultivation on the BC material properties, such as crystallinity, swelling ratio, and moisture content. Applying specific enzymes for converting inaccessible, raw-form carbon sources to the culture medium of Komagateibacter xylinus opens a simple way to use various oligo- and polysaccharides acquired from many kinds of biomass sources in the BC production process.
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