title: Co-refolding of two peptide fragments derived from Agrobacterium tumefaciens beta-glucosidase with catalytic activity.
abstract: Four sites of the non-homologous region (coding amino acid residues of 347, 421, 466 and 533) of a gene were randomly selected for splitting to investigate the function of beta-glucosidase from Agrobacterium tumefaciens in the co-refolding of peptides into the catalytically active enzyme. As a result of gene splitting, four N- and C-terminal domain peptides were obtained as insoluble inclusion bodies. No catalytic activity was observed when these fragments refolded individually. However, a considerable amount of activity was restored when the two fragments derived from N- and C- terminal peptides were co-refolded together. The deletion of amino acid residues in the non-homologous region resulted in a complete loss of enzyme activity, which suggests that truncation of amino acids in this region strongly affects the co-refolding ability of the enzyme to maintain activity.
authors.affiliation: Enzyme Laboratory, National Food Research Institute, 2-1-12 Kannondai, Tsukuba 305-8642, Japan. bjkim@mail.nih.gov
authors.forename: Bong-Jo
authors.initials: BJ
authors.lastname: Kim
authors.forename: Selanere L
authors.initials: SL
authors.lastname: Mangala
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Hayashi
mesh_terms: D016960:Agrobacterium tumefaciens / Q000201:enzymology*; D000438:Alcohols / Q000494:pharmacology; D002384:Catalysis; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D020673:Protein Renaturation / Q000187:drug effects; D012965:Sodium Chloride / Q000494:pharmacology; D013379:Substrate Specificity; D013696:Temperature; D001617:beta-Glucosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D000438:Alcohols; D010446:Peptide Fragments; D012965:Sodium Chloride; D001617:beta-Glucosidase