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α-Glucosidase from a strain of deep-sea Geobacillus: a potential enzyme for the biosynthesis of complex carbohydrates

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Abstract

An α-glucosidase from Geobacillus sp. strain HTA-462, one of the deepest sea bacteria isolated from the sediment of the Mariana Trench, was purified to homogeneity and estimated to be a 65-kDa protein by SDS-PAGE. At low ion strength, the enzyme exists in the homodimeric form (130 kDa). It is a thermo- and alkaline-stable enzyme with a half-life of 13.4 h and a maximum hydrolytic activity at 60°C and pH 9.0 in 15 mM glycine–NaOH buffer. The enzyme exclusively hydrolyzed α-1,4-glycosidic linkages of oligosaccharides in an exo-type manner. The enzyme had an overwhelming transglycosylation activity and glycosylated various non-sugar molecules when maltose was used as a sugar donor. It converted maltose to isomaltose. The gene encoding the enzyme was cloned and sequenced. The recombinant enzyme could be extracellularly overproduced by Bacillus subtilis harboring its gene and preserved the primary properties of the native enzyme. Site-directed mutagenesis experiments showed that Asp98 is essential for the enzyme activity in addition to Asp199, Asp326, and Glu256.

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Acknowledgements

We sincerely thank Dr. H. Takami for providing us with the organisms used in this study.

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Authors and Affiliations

  1. Japan Agency for Marine–Earth Science and Technology (JAMSTEC), 2-15 Natsushima, Yokosuka, Kanagawa, 237-0061, Japan

    Vo Si Hung, Yuji Hatada, Saori Goda, Jie Lu, Yuko Hidaka, Zhijun Li, Masatake Akita, Yukari Ohta, Susumu Ito & Koki Horikoshi

  2. Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Kita-9, Nishi-9, Kita-ku, Sapporo, 060-8589, Japan

    Kenji Watanabe & Hirokazu Matsui

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  1. Vo Si Hung
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  2. Yuji Hatada
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  3. Saori Goda
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  4. Jie Lu
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  5. Yuko Hidaka
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  7. Masatake Akita
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  11. Susumu Ito
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  12. Koki Horikoshi
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Correspondence to Vo Si Hung.

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Hung, V.S., Hatada, Y., Goda, S. et al. α-Glucosidase from a strain of deep-sea Geobacillus: a potential enzyme for the biosynthesis of complex carbohydrates. Appl Microbiol Biotechnol 68, 757–765 (2005). https://doi.org/10.1007/s00253-005-1977-3

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  • DOI: https://doi.org/10.1007/s00253-005-1977-3

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