title: Effects of truncation at the non-homologous region of a family 3 beta-glucosidase from Agrobacterium tumefaciens.
abstract: The function of the non-homologous region of a family 3 beta-glucosidase from Agrobacterium tumefaciens (Cbg1) was studied by analyzing the properties of mutant enzymes that have internal truncated amino acid sequences in the region. Five truncated mutants named Cbg1-d4, Cbg1-d31, Cbg1-d62, Cbg1-d89, and Cbg1-d119 having deletions of 4, 31, 62, 89, and 119 amino acid residues starting from Phe417, respectively, were expressed in Escherichia coli and purified. All the mutants exhibited beta-glucosidase activity, indicating that the non-homologous region was not essential for the activity. The truncation caused thermal instability, decrease in pK(a) of the proton donor residue (Glu616), and deficient transglycosylation activity. The thermal stability and the pK(a) of Glu616 were partially recovered with longer truncation, suggesting that the truncation perturbed the structure and that their presence in the region was not essential. The main role of the non-homologous region could be formation of a hydrophobic atmosphere at the acceptor site to make the enzyme suitable for hydrolyzing hydrophobic glucosides.
authors.affiliation: Enzyme Laboratory, National Food Research Institute, Ibaraki, Japan.
authors.forename: Li
authors.initials: L
authors.lastname: Ying
authors.forename: Motomitsu
authors.initials: M
authors.lastname: Kitaoka
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Hayashi
mesh_terms: D016960:Agrobacterium tumefaciens / Q000201:enzymology* / Q000235:genetics; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004795:Enzyme Stability / Q000235:genetics; D006031:Glycosylation; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D017384:Sequence Deletion / Q000235:genetics; D001617:beta-Glucosidase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D001617:beta-Glucosidase