Extracellular polymeric matrices (EPMs) are complex mixtures of carbohydrates, proteins and lipid derivatives secreted by certain bacteria that serve as a source of energy for the gut microbiome. The breakdown of EPMs in the gut leads to the production of short-chain fatty acids (SCFAs) through microbial fermentation. SCFAs are essential in maintaining polymicrobial colonisation and protection against gut-metabolic disorders. This is the first study utilising and characterising the Bacillus spizizenii VSMKU0112 derived EPM. The composition, structural and functional moieties were analysed using Fourier transform infrared spectroscopy and 1H and 13C NMR, confirming that the EPM major component was a heteropolysaccharide derivative with beta-linked fructose, glucose and ribose and named beta-glucoribofructan (beta-GRF). The beta-GRF was evaluated for antioxidant activity, with scavenging and reducing power of 29.22 +/- 0.1 and 21.11 +/- 1.2 mu g mL-1, respectively. Furthermore, antidiabetic activity showed reduced half-maximal inhibition concentration values at alpha-amylase 2.93 +/- 0.21, beta-galactosidase 2.35 +/- 0.32, dipeptidyl peptidase 10.4 +/- 0.43 mu g mL-1. Anti-inflammatory activity was observed at 20-30 mu g mL-1. The antilipidemic properties exhibited inhibitory effects of up to 35% in cholesterol esterase and 55% in pancreatic lipase. beta-GRF was then subjected to artificial simulated gastrointestinal digestion using human gut microbiota from healthy individuals. The post-digestive substrate was analysed for SCFAs using gas chromatography-mass spectrometry (GC-MS/MS), indicating the presence of various SCFAs and long-chain fatty acids. Moreover, in vitro studies using gut homogenates from inflammatory bowel disease groups showed that beta-GRF treatment enhanced the SCFA production by the improvement of microbial colonisation, which was studied through sequencing the V3-V4 region of the 16S rRNA gene and total metabolome analysis by GC-MS/MS. Therefore, beta-GRF utilisation highlights a potential impact in the treatment of metabolic-regulated inflammatory bowel disease. (c) 2025 Society of Chemical Industry.