Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning and Characterization of Glycogen-Debranching Enzyme from Hyperthermophilic Archaeon Sulfolobus shibatae

  • Van, Trinh Thi Kim (Research Institute of Food and Nutritional Sciences and Department of Food and Nutrition, Brain Korea 21 Project, Yonsei University) ;
  • Ryu, Soo-In (Research Institute of Food and Nutritional Sciences and Department of Food and Nutrition, Brain Korea 21 Project, Yonsei University) ;
  • Lee, Kyung-Ju (Research Institute of Food and Nutritional Sciences and Department of Food and Nutrition, Brain Korea 21 Project, Yonsei University) ;
  • Kim, Eun-Ju (Research Institute of Food and Nutritional Sciences and Department of Food and Nutrition, Brain Korea 21 Project, Yonsei University) ;
  • Lee, Soo-Bok (Research Institute of Food and Nutritional Sciences and Department of Food and Nutrition, Brain Korea 21 Project, Yonsei University)
  • Published : 2007.05.31
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Abstract

A gene encoding a putative glycogen-debranching enzyme in Sulfolobus shibatae(abbreviated as SSGDE) was cloned and expressed in Escherichia coli. The recombinant enzyme was purified to homogeneity by heat treatment and Ni-NTA affinity chromatography. The recombinant SSGDE was extremely thermostable, with an optimal temperature at $85^{\circ}C$. The enzyme had an optimum pH of 5.5 and was highly stable from pH 4.5 to 6.5. The substrate specificity of SSGDE suggested that it possesses characteristics of both amylo-1,6-glucosidase and $\alpha$-1,4-glucanotransferase. SSGDE clearly hydrolyzed pullulan to maltotriose, and $6-O-\alpha-maltosyl-\beta-cyclodextrin(G2-\beta-CD)$ to maltose and $\beta$-cyclodextrin. At the same time, SSGDE transferred maltooligosyl residues to the maltooligosaccharides employed, and maltosyl residues to $G2-\beta-CD$. The enzyme preferentially hydrolyzed amylopectin, followed in a decreasing order by glycogen, pullulan, and amylose. Therefore, the present results suggest that the glycogen-debranching enzyme from S. shibatae may have industrial application for the efficient debranching and modification of starch to dextrins at a high temperature.

Keywords

References

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