Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Enzymatic Characterization and Substrate Specificity of Thermostable $\beta-Glycosidase$ from Hyperthermophilic Archaea, Sulfolobus shibatae, Expressed in E. coli

  • Park, Na-Young (Department of Food Science and Biotechnology, Graduate School of Biotechnology & Institute of Life Sciences and Resources, KyungHee University) ;
  • Cha, Jae-Ho (Department of Microbiology, Pusan National University) ;
  • Kim, Dae-Ok (Department of Food Science and Biotechnology, Graduate School of Biotechnology & Institute of Life Sciences and Resources, KyungHee University) ;
  • Park, Cheon-Seok (Department of Food Science and Biotechnology, Graduate School of Biotechnology & Institute of Life Sciences and Resources, KyungHee University)
  • Published : 2007.03.31
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Abstract

Enzymatic properties and substrate specificity of recombinant $\beta-glycosidases$ from a hyperthermophilic archaeon, Sulfolobus shibatae (rSSG), were analyzed. rSSG showed its optimum temperature and pH at $95^{\circ}C$ and pH 5.0, respectively. Thermal inactivation of rSSG showed that its half-life of enzymatic activity at $75^{\circ}C$ was 15 h whereas it drastically decreased to 3.9 min at $95^{\circ}C$. The addition of 10 mM of $MnCl_2$ enhanced the hydrolysis activity of rSSG up to 23% whereas most metal ions did not show any considerable effect. Dithiothreitol (DTT) and 2-mercaptoethanol exhibited significant influence on the increase of the hydrolysis activity of rSSG rSSG apparently preferred laminaribiose $(\beta1\rightarrow3Glc)$, followed by sophorose $(\beta1\rightarrow2Glc)$, gentiobiose $(\beta1\rightarrow6Glc)$, and cellobiose $(\beta1\rightarrow4Glc)$. Various. intermolecular transfer products were formed by rSSG in the lactose reaction, indicating that rSSG prefers lactose as a good acceptor as well as a donor. The strong intermolecular transglycosylation activity of rSSG can be applied in making functional oligosaccharides.

Keywords

References

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