title: Effects of the N-terminal and C-terminal domains of Meiothermus ruber CBS-01 trehalose synthase on thermostability and activity.
abstract: Numerous trehalose synthases (TreS) from thermophilic microorganisms have extra C-terminal domains. To determine the function of the N- and C-terminal domains of TreS from the thermophilic bacterium Meiothermus ruber CBS-01, the two domains were expressed. From the findings, the N-terminal domain from M. ruber was not active when compared with that from Thermus thermophilus, which had been studied previously. The circular dichroism spectrum showed that the secondary structure of N-terminal domain from M. ruber underwent a greater change than that of C terminus. In addition, the N-terminal domain from T. thermophilus and C terminus from M. ruber were fused. The fusion protein TSTtMr was more efficient and thermostable than the TreS from M. ruber. The N-terminal domain from M. ruber and C terminus from T. thermophilus were fused. The optimum temperature and thermostability of fusion protein TSMrTt were similar to the TreS from M. ruber. It was presumed that aside from the C-terminal domain, the N-terminal domain of TreS from thermophilic bacteria could influence thermostability. For the TreS from M. ruber, the mutant protein R392F led to a complete loss in activity, and R392A showed a sharp decrease in activity.
authors.affiliation: Key Laboratory of Molecular Microbiology and Technology, Department of Microbiology, Nankai University, Ministry of Education, Tianjin, 300071, People's Republic of China.
authors.forename: Yufan
authors.initials: Y
authors.lastname: Wang
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Wenwen
authors.initials: W
authors.lastname: Wang
authors.forename: Yanchao
authors.initials: Y
authors.lastname: Liu
authors.forename: Laijun
authors.initials: L
authors.lastname: Xing
authors.forename: Mingchun
authors.initials: M
authors.lastname: Li
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004795:Enzyme Stability; D005964:Glucosyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006095:Gram-Positive Cocci / Q000201:enzymology* / Q000235:genetics; D006358:Hot Temperature; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D005964:Glucosyltransferases; C099692:trehalose synthase