title: Purification and characterization of thermostable pectate lyase with protopectinase activity from thermophilic Bacillus sp. TS 47.
abstract: A strain of thermophilic bacterium, Bacillus sp., with pectolytic activity has been isolated. It produced an extracellular endo-polygalacturonate trans-eliminase (PL, EC 4.2.2.1) when grown at 60 degrees C on a medium containing polygalacturonate (PGA). The PL was purified by hydrophobic, cation exchange, and size exclusion column chromatographies. The molecular mass of the enzyme was 50 kDa by SDS-PAGE. The isoelectric point of the enzyme was pH 5.3. The enzyme had a half-life of 13 and 1 h at 65 and 70 degrees C, respectively, and showed optimal activity around at 70 degrees C and pH 8.0. It had protopectinase activity, besides PL activity, on lemon protopectin and cotton fibers. The first 20 amino acids sequence of the enzyme had significant similarity with that of PL from methophilic Bacillus subtilis, with 50% identity.
authors.affiliation: Department of Food Science and Nutrition, Faculty of Agriculture, Kinki University, Nakamachi, Nara, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Takao
authors.forename: T
authors.initials: T
authors.lastname: Nakaniwa
authors.forename: K
authors.initials: K
authors.lastname: Yoshikawa
authors.forename: T
authors.initials: T
authors.lastname: Terashita
authors.forename: T
authors.initials: T
authors.lastname: Sakai
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology* / Q000378:metabolism; D004795:Enzyme Stability; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000378:metabolism; D008969:Molecular Sequence Data; D011133:Polysaccharide-Lyases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D008670:Metals; D006026:Glycoside Hydrolases; C083413:protopectinase; D011133:Polysaccharide-Lyases; C023911:pectate lyase