İspirli H.Şimşek Ö.Skory C.Sağdıç O.Dertli E.20.04.20192019-04-2020.04.20192019-04-2020190141-8130https://dx.doi.org/10.1016/j.ijbiomac.2018.12.050https://hdl.handle.net/20.500.12403/310A wide number of Lactic Acid Bacteria (LAB) species produce ?-glucans with their ability to synthesize glucansucrases (GS) which use sucrose as substrate for the glucan production. Recently another group of enzymes in LAB gained special interest for their ability to produce ?-glucans targeting the substrates containing ?1-4-linkages and synthesizing new (?1-6) or (?1-3)–linkages as ??glucanotransferases. In this study, a putative 4,6???glucanotransferase (GTFB) from sourdough isolate Lactobacillus reuteri E81 was identified and expressed in Escherichia coli. The biochemical characterization of the GTFB-E81 confirmed its function as it cleaved the ?1-4-linkages in different substrates and produced new gluco-oligomers/polymers containing ?1-6 linkages together with the ?1-4-linkages detected by NMR analysis. GTFB-E81 produced malto-oligosaccharides targeting maltose and maltoheptaose as substrates with up to DP 8 detected by TLC and ESI-MS/MS analysis. The functional roles of these malto-oligosaccharides were determined by testing their immune-modulatory functions in HT29 cells and they triggered the production of anti-inflammatory 1L-4 and pro-inflammatory IL-12 cytokines. © 2018 Elsevier B.V.eninfo:eu-repo/semantics/closedAccess4,6???GlucanotransferaseImmune-modulatory functionsMalto-oligosaccharides4,6 alpha glucanotransferaseglycosyltransferaseinterleukin 12interleukin 4maltooligosaccharidemaltoseoligomeroligosaccharidepolymerunclassified drugArticlebacterium identificationbacterium isolatebiochemical analysiscarbohydrate synthesiscontrolled studyelectrospray mass spectrometryenzyme substrateEscherichia coliHT-29 cell linehumanhuman cellimmunomodulationLactobacillus reuteriLactobacillus reuteri E81nonhumannuclear magnetic resonancenucleotide sequenceprotein cleavageprotein cross linkingprotein functionthin layer chromatography4,6???GlucanotransferaseImmune-modulatory functionsMalto-oligosaccharides4,6 alpha glucanotransferaseglycosyltransferaseinterleukin 12interleukin 4maltooligosaccharidemaltoseoligomeroligosaccharidepolymerunclassified drugArticlebacterium identificationbacterium isolatebiochemical analysiscarbohydrate synthesiscontrolled studyelectrospray mass spectrometryenzyme substrateEscherichia coliHT-29 cell linehumanhuman cellimmunomodulationLactobacillus reuteriLactobacillus reuteri E81nonhumannuclear magnetic resonancenucleotide sequenceprotein cleavageprotein cross linkingprotein functionthin layer chromatographyArticle1241213121910.1016/j.ijbiomac.2018.12.050305292032-s2.0-85057749968Q1WOS:000456899300127Q1