Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Molecular Cloning and Characterization of Trehalose Biosynthesis Genes from Hyperthermophilic Archaebacterium Metallosphaera hakonesis

  • Seo, Ju-Seok (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University) ;
  • An, Ju-Hee (Department of Food and Nutrition, Seowon University) ;
  • Baik, Moo-Yeol (Department of Food Science and Biotechnology, Graduate School of Biotechnology and Institute of Life Science & Resources, Kyung Hee University) ;
  • Park, Cheon-Seok (Department of Food Science and Biotechnology, Graduate School of Biotechnology and Institute of Life Science & Resources, Kyung Hee University) ;
  • Cheong, Jong-Joo (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University) ;
  • Moon, Tae-Wha (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University) ;
  • Park, Kwan-Hwa (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University) ;
  • Choi, Yang-Do (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University) ;
  • Kim, Chung-Ho (Department of Food and Nutrition, Seowon University)
  • Published : 2007.01.31
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Abstract

The trehalose $({\alpha}-D-glucopyranosyl-[1,1]-{\alpha}-D-glucopyranose)$ biosynthesis genes MhMTS and MhMTH, encoding a maltooligosyltrehalose synthase (MhMTS) and a maltooligosyltrehalose trehalohydrolase (MhMTH), respectively, have been cloned from the hyperthermophilic archaebacterium Metallosphaera hakonesis. The ORF of MhMTS is 2,142 bp long, and encodes 713 amino acid residues constituting a 83.8 kDa protein. MhMTH is 1,677 bp long, and encodes 558 amino acid residues constituting a 63.7 kDa protein. The deduced amino acid sequences of MhMTS and MhMTH contain four regions highly conserved for MTSs and three for MTHs that are known to constitute substrate-binding sites of starch-hydrolyzing enzymes. Recombinant proteins obtained by expressing the MhMTS and MhMTH genes in E. coli catalyzed a sequential reaction converting maltooligosaccharides to produce trehalose. Optimum pH of the MhMTS/MhMTH enzyme reaction was around 5.0 and optimum temperature was around 70 C. Trehalose-producing activity of the MhMTS/ MhMTH was notably stable, retaining 80% of the activity after preincubation of the enzyme mixture at $70^{\circ}C$ for 48 h, but was gradually abolished by incubating at above $85^{\circ}C$. Addition of thermostable $4-{\alpha}-glucanotransferase$ increased the yield of trehalose production from maltopentaose by 10%. The substrate specificity of the MhMTS/MhMTH-catalyzed reaction was extended to soluble starch, the most abundant maltodextrin in nature.

Keywords

References

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