2024-03-29T07:05:29Zhttps://eprints.lib.hokudai.ac.jp/dspace-oai/requestoai:eprints.lib.hokudai.ac.jp:2115/876342023-01-13T17:17:39Zhdl_2115_20046hdl_2115_138A practical approach to producing isomaltomegalosaccharide using dextran dextrinase from Gluconobacter oxydans ATCC 11894Lang, WeeranuchKumagai, YuyaSadahiro, JuriSaburi, WataruSarnthima, RakrudeeTagami, TakayoshiOkuyama, MasayukiMori, HaruhideSakairi, NobuoKim, DomanKimura, AtsuoGluconobacter oxydansDextran dextrinaseDextranIsomaltomegalosaccharideChimeric glucosaccharide610Dextran dextrinase (DDase) catalyzes formation of the polysaccharide dextran from maltodextrin. During the synthesis of dextran, DDase also generates the beneficial material isomaltomegalosaccharide (IMS). The term megalosaccharide is used for a saccharide having DP=10-100 or 10-200 (DP, degree of polymerization). IMS is a chimeric glucosaccharide comprising alpha-(1 -> 6)- and alpha-(1 -> 4)-linked portions at the nonreducing and reducing ends, respectively, in which the alpha-(1 -> 4)-glucosyl portion originates from maltodextrin of the substrate. In this study, IMS was produced by a practical approach using extracellular DDase (DDext) or cell surface DDase (DDsur) of Gluconobacter oxydans ATCC 11894. DDsur was the original form, so we prepared DDext via secretion from intact cells by incubating with 0.5% G6/G7 (maltohexaose/maltoheptaose); this was followed by generation of IMS from various concentrations of G6/G7 substrate at different temperatures for 96 h. However, IMS synthesis by DDext was limited by insufficient formation of alpha-(1 -> 6)-glucosidic linkages, suggesting that DDase also catalyzes elongation of alpha-(1 -> 4)-glucosyl chain. For production of IMS using DDsur, intact cells bearing DDsur were directly incubated with 20% G6/G7 at 45 degrees C by optimizing conditions such as cell concentration and agitation efficiency, which resulted in generation of IMS (average DP =14.7) with 61% alpha-(1 -> 6)-glucosyl content in 51% yield. Increases in substrate concentration and agitation efficiency were found to decrease dextran formation and increase IMS production, which improved the reaction conditions for DDext. Under modified conditions (20% G6/G7, agitation speed of 100 rpm at 45 degrees C), DDext produced IMS (average DP =14.5) with 65% alpha-(1 -> 6)-glucosyl content in a good yield of 87%.SpringerJournal Articleapplication/pdfhttp://hdl.handle.net/2115/87634https://eprints.lib.hokudai.ac.jp/dspace/bitstream/2115/87634/1/Applied%20microbiology%20and%20biotechnology.pdf0175-7598Applied microbiology and biotechnology1066896982022-01-13enginfo:doi/10.1007/s00253-021-11753-6This version of the article has been accepted for publication, after peer review (when applicable) and is subject to Springer Nature’s AM terms of use, but is not the Version of Record and does not reflect post-acceptance improvements, or any corrections. The Version of Record is available online at: http://dx.doi.org/10.1007/s00253-021-11753-6author