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Biosynthesis of Glucosyl Glycerol, a Compatible Solute, Using Intermolecular Transglycosylation Activity of Amylosucrase from Methylobacillus flagellatus KT

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Abstract

A putative α-amylase gene (accession number, CP000284) of Methylobacillus flagellatus KT ATCC51484 was cloned in Escherichia coli, and its gene product was expressed and characterized. The purified recombinant enzyme (MFAS) displayed a typical amylosucrase (ASase) activity by the demonstration of multiple activities of hydrolysis, isomerization, and polymerization although it was designated as an α-amylase. The optimal reaction temperature and pH for the sucrose hydrolysis activity of MFAS were determined to be 45 °C and pH 8.5, respectively. MFAS has relatively high thermostable characteristics compared with other ASases, as demonstrated by a half-life of 19.3 min at 50 °C. MFAS also showed polymerization activity using sucrose as a sole substrate. Glycerol was transglycosylated by the intermolecular transglycosylation activity of MFAS. Two major products were observed by thin-layer chromatography and isolated by paper chromatography and recycling HPLC. Using 1H and 13C NMR, their chemical structures were determined to be (2S)-1-O-α-d-glucosyl-glycerol or (2R)-1-O-α-d-glucosyl-glycerol and 2-O-α-d-glucosyl-glycerol, in which a glucose molecule is linked to glycerol via an α-glycosidic linkage.

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Acknowledgment

This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST) (No. 2013–031011) and by Business for Cooperative R&D between Industry, Academy, and Research Institute funded Korea Small and Medium Business Administration in 2013 (Grant No. C0123542).

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Authors and Affiliations

  1. Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin, 446-701, South Korea

    Jin-Woo Jeong, Dong-Ho Seo, Jong-Hyun Jung, Ji-Hae Park, Nam-In Baek & Cheon-Seok Park

  2. Food Research Institute and School of Food and Life Science and Biohealth Products Research Center, Inje University, Gimhae, 621-749, South Korea

    Myo-Jeong Kim

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  1. Jin-Woo Jeong
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  2. Dong-Ho Seo
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  3. Jong-Hyun Jung
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Correspondence to Cheon-Seok Park.

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Jin-Woo Jeong and Dong-Ho Seo contributed equally.

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Jeong, JW., Seo, DH., Jung, JH. et al. Biosynthesis of Glucosyl Glycerol, a Compatible Solute, Using Intermolecular Transglycosylation Activity of Amylosucrase from Methylobacillus flagellatus KT. Appl Biochem Biotechnol 173, 904–917 (2014). https://doi.org/10.1007/s12010-014-0889-z

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  • DOI: https://doi.org/10.1007/s12010-014-0889-z

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