title: Purification, characterization, and gene analysis of cellulase (Cel8A) from Lysobacter sp. IB-9374.
abstract: An enzyme that has both beta-1,4-glucanase and chitosanase activities was found in the culture medium of the soil bacterium Lysobacter sp. IB-9374, a high lysyl endopeptidase-producing strain. The enzyme was purified to homogeneity from the culture filtrate using five purification steps and designated Cel8A. The purified Cel8A had a molecular mass of 41 kDa, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A pH optimum of 5.0 was found for the beta-1,4-glucanase activity, and pH optima of 5.0 and 7.0 were found for the chitosanase activity. Nucleotide sequencing of the Cel8A gene yielded a deduced amino acid sequence that comprises a 33-amino acid, N-terminal signal peptide and a mature enzyme consisting of a 381-residue polypeptide with a predicted molecular mass of 41,241 Da. The amino acid sequence of the Cel8A, which contains the catalytic module of glycosyl hydrolase family 8, is homologous to beta-1,3-1,4-D-glucanase from Bacillus circulans WL-12 and endoglucanase N-257 from B. circulans KSM-N257.
authors.affiliation: United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Japan.
authors.forename: Jiro
authors.initials: J
authors.lastname: Ogura
authors.forename: Atsushi
authors.initials: A
authors.lastname: Toyoda
authors.forename: Taisuke
authors.initials: T
authors.lastname: Kurosawa
authors.forename: Ai Leng
authors.initials: AL
authors.lastname: Chong
authors.forename: Shigeru
authors.initials: S
authors.lastname: Chohnan
authors.forename: Takeharu
authors.initials: T
authors.lastname: Masaki
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D020134:Catalytic Domain; D002480:Cellulase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases; D006863:Hydrogen-Ion Concentration; D053528:Lysobacter / Q000201:enzymology / Q000235:genetics*; D008970:Molecular Weight; D021382:Protein Sorting Signals; D020539:Sequence Analysis, Protein; D017385:Sequence Homology; D012988:Soil Microbiology
chemical_list: D001426:Bacterial Proteins; D021382:Protein Sorting Signals; D006026:Glycoside Hydrolases; C019868:chitosanase; D002480:Cellulase