biostudies-literatureWienberg FBundesministerium für Bildung und ForschungRheinische Friedrich-Wilhelms-Universität Bonn144https://www.ebi.ac.uk/biostudies/studies/S-EPMC9666576biostudies-literatureUnknown12(1)Due to the health-promoting effects and functional properties of inulin-type fructooligosaccharides (I-FOS), the global market for I-FOS is constantly growing. Hence, there is a continuing demand for new, efficient biotechnological approaches for I-FOS production. In this work, crude inulosucrase InuGB-V3 from Lactobacillus gasseri DSM 20604 was used to synthesize I-FOS from sucrose. Supplementation with 1 mM CaCl₂, a pH of 3.5-5.5, and an incubation temperature of 40 °C were found to be optimal production parameters at which crude inulosucrase showed high conversion rates, low sucrose hydrolysis, and excellent stability over 4 days. The optimal process conditions were employed in cell-free bioconversion reactions. By elevating the substrate concentration from 570 to 800 g L<supAMB ExpressHigh-yield production and purification of prebiotic inulin-type fructooligosaccharides.PMC9666576FKZ 031B1054AWienberg FDeppenmeier UHovels MfalseHigh-yield production and purification of prebiotic inulin-type fructooligosaccharides.Due to the health-promoting effects and functional properties of inulin-type fructooligosaccharides (I-FOS), the global market for I-FOS is constantly growing. Hence, there is a continuing demand for new, efficient biotechnological approaches for I-FOS production. In this work, crude inulosucrase InuGB-V3 from Lactobacillus gasseri DSM 20604 was used to synthesize I-FOS from sucrose. Supplementation with 1 mM CaCl₂, a pH of 3.5-5.5, and an incubation temperature of 40 °C were found to be optimal production parameters at which crude inulosucrase showed high conversion rates, low sucrose hydrolysis, and excellent stability over 4 days. The optimal process conditions were employed in cell-free bioconversion reactions. By elevating the substrate concentration from 570 to 800 g L<sup2022-01-01T00:00:00Z2022 Nov2025-04-19T13:25:07.213Z2025-04-19T13:25:07.213ZS-EPMC96665763638021310.1186/s13568-022-01485-9