Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning and Overexpression of 4-${\alpha}$-Glucanotransferase from Thermus brockianus (TBGT) in E. coli

  • Bang, Bo-Young (Department of Food Science & Biotechnology, Graduate School of Biotechnology and Institute of Life Science & Resources, KyungHee University) ;
  • Kim, Han-Jo (Department of Food Science & Biotechnology, Graduate School of Biotechnology and Institute of Life Science & Resources, KyungHee University) ;
  • Kim, Hae-Yeong (Department of Food Science & Biotechnology, Graduate School of Biotechnology and Institute of Life Science & Resources, KyungHee University) ;
  • Baik, Moo-Yeol (Department of Food Science & Biotechnology, Graduate School of Biotechnology and Institute of Life Science & Resources, KyungHee University) ;
  • Ahn, Soon-Cheol (Department of Microbiology and Immunology, College of Medicine, Pusan National University) ;
  • Kim, Chung-Ho (Department of Food and Nutrition, Seowon University) ;
  • Park, Cheon-Seok (Department of Food Science & Biotechnology, Graduate School of Biotechnology and Institute of Life Science & Resources, KyungHee University)
  • Published : 2006.11.30
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Abstract

A gene corresponding to 4-${\alpha}$-glucanotransferase (${\alpha}GTase$) was cloned from the thermophilic bacterium Thermus brockianus. The nucleotide sequence analysis showed that the ${\alpha}GTase$ gene is composed of 1,503 nucleotides and encodes a polypeptide that is 500 amino acids long with a calculated molecular mass of 57,221 Da. The deduced amino acid sequences of Thermus brockianus ${\alpha}GTase$ (TBGT) exhibited a high level of similarity to the amino acid sequence of ${\alpha}GTase$ of Thermus thermophilus (86%), but low level of homology to that of E. coli (26%). The TBGT gene was overexpressed in E. coli BL21, and the corresponding recombinant enzyme was efficiently purified by Ni-NTA affinity chromatography. The enzymatic characteristics revealed that optimal pH and temperature were pH 6 and $70^{\circ}C$, respectively. Most interestingly, TBGT reacted with small oligosaccharides, especially maltotriose, to form various maltooligosaccharides by using its disproportionation activity.

Keywords

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