Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Recombinant Expression and Characterization of Thermoanaerobacter tengcongensis Thermostable $\alpha$-Glucosidase with Regioselectivity for High-Yield Isomaltooligosaccharides Synthesis

  • Zhou, Cheng (State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences) ;
  • Xue, Yanfen (State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences) ;
  • Zhang, Yueling (Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences) ;
  • Zeng, Yan (State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences) ;
  • Ma, Yanhe (State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences)
  • Published : 2009.12.31
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Abstract

A novel thermostable $\alpha$-glucosidase (TtGluA) from Thermoanaerobacter tengcongensis MB4 was successfully expressed in E. coli and characterized. The TtgluA gene contained 2,253 bp, which encodes 750 amino acids. The native TtGluA was a trimer with monomer molecular mass of 89 kDa shown by SDS-PAGE. The purified recombinant enzyme showed hydrolytic activity on maltooligosaccharides, p-nitrophenyl-$\alpha$-D-glucopyranide, and dextrin with an exotype cleavage manner. TtGluA showed preference for short-chain maltooligosaccharides and the highest specific activity for maltose of 3.26 units/mg. Maximal activity was observed at $60^{\circ}C$ and pH 5.5. The half-life was 2 h at $60^{\circ}C$. The enzyme showed good tolerance to urea and SDS but was inhibited by Tris. When maltose with the concentration over 50 mM was used as substrate, TtGluA was also capable of catalyzing transglycosylation to produce $\alpha$-1,4-linked maltotriose and $\alpha$-1,6-linked isomaltooligosaccharides. More importantly, TtGluA showed exclusive regiospecificity with high yield to produce $\alpha$-1,6-linked isomaltooligosaccharides when the reaction time extended to more than 10 h.

Keywords

References

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