Abstract
Dextran 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 α-(1 → 6)- and α-(1 → 4)-linked portions at the nonreducing and reducing ends, respectively, in which the α-(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 α-(1 → 6)-glucosidic linkages, suggesting that DDase also catalyzes elongation of α-(1 → 4)-glucosyl chain. For production of IMS using DDsur, intact cells bearing DDsur were directly incubated with 20% G6/G7 at 45 °C by optimizing conditions such as cell concentration and agitation efficiency, which resulted in generation of IMS (average DP = 14.7) with 61% α-(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 °C), DDext produced IMS (average DP = 14.5) with 65% α-(1 → 6)-glucosyl content in a good yield of 87%.
Key points
• Beneficial IMS was produced using thermostabilized DDase.
• Optimum conditions for reduced dextran formation were successfully determined.
• A practical approach was established to provide IMS with a great yield of 87%.
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The data that support the findings of this study are available from the corresponding authors on reasonable request.
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Acknowledgements
We would like to thank Nihon Shokuhin Kako (Tokyo, Japan) for kindly gifting us both products of Fujioligo G6 and G7 and Fujioligo G3.
Funding
This study was supported partially by a Program for Promotion of Basic and Applied Research for Innovations in Biooriented Industry (BRAIN, Japan; Grant No. 26062B) and the Japan Society for the Promotion of Science KAKENHI Grant Nos. 17H03801 and 19KK0147.
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WL and AK conceived and coordinated the study. WL, YK, JS, WS, and RS conducted experiments. WL, YK, JS, WS, RS, TT, MO, HM, NS, DK, and AK analyzed the data. WL and AK wrote the manuscript. All authors read and approved the final manuscript.
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Lang, W., Kumagai, Y., Sadahiro, J. et al. A practical approach to producing isomaltomegalosaccharide using dextran dextrinase from Gluconobacter oxydans ATCC 11894. Appl Microbiol Biotechnol 106, 689–698 (2022). https://doi.org/10.1007/s00253-021-11753-6
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DOI: https://doi.org/10.1007/s00253-021-11753-6