title: Analysis of bgl operon structure and characterization of beta-glucosidase from Pectobacterium carotovorum subsp. carotovorum LY34.
abstract: A putative bgl operon of Pectobacterium carotovorum subsp. carotovorum LY34 (Pcc LY34) was isolated. Sequence analysis of the 5,557 bp cloned DNA fragment (accession no. AY542524) showed three open reading frames (bglT, bglP, and bglB) predicted to encode 287, 633, and 468 amino acid proteins respectively. BglT and BglP ORFs show high similarity to that of the Pectobacterium chrysanthemi ArbG antiterminator and ArbF permease respectively. Also, the latter contains most residues important for phosphotransferase activity. The amino acid sequence of BglB showed high similarity to various beta-glucosidases and is a member of glycosyl hydrolase family 1. The purified BglB enzyme hydrolyzed salicin, arbutin, pNPG, and MUG. The molecular weight of the enzyme was estimated to be 53,000 Da by SDS-PAGE. The purified beta-glucosidase exhibited maximal activity at pH 7.0 and 40 degrees C, and its activity was enhanced in the presence of Mg(2+). Two glutamate residues (Glu(173) and Glu(362)) were found to be essential for enzyme activity.
authors.affiliation: Division of Applied Life Science, Gyeongsang National University, Chinju, Korea.
authors.forename: Chang Long
authors.initials: CL
authors.lastname: An
authors.forename: Woo Jin
authors.initials: WJ
authors.lastname: Lim
authors.forename: Su Young
authors.initials: SY
authors.lastname: Hong
authors.forename: Eun Ju
authors.initials: EJ
authors.lastname: Kim
authors.forename: Eun Chule
authors.initials: EC
authors.lastname: Shin
authors.forename: Min Keun
authors.initials: MK
authors.lastname: Kim
authors.forename: Jong Reoul
authors.initials: JR
authors.lastname: Lee
authors.forename: Sang Ryeol
authors.initials: SR
authors.lastname: Park
authors.forename: Jong Gyu
authors.initials: JG
authors.lastname: Woo
authors.forename: Yong Pyo
authors.initials: YP
authors.lastname: Lim
authors.forename: Han Dae
authors.initials: HD
authors.lastname: Yun
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D003360:Cosmids; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D015723:Gene Library; D026901:Membrane Transport Proteins / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009876:Operon / Q000235:genetics*; D044043:Pectobacterium / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D011995:Recombination, Genetic; D013696:Temperature; D001617:beta-Glucosidase / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D026901:Membrane Transport Proteins; D008670:Metals; D001617:beta-Glucosidase