title: Purification and characterization of novel transglutaminase from Bacillus subtilis spores.
abstract: Transglutaminase activity was detected in suspensions of purified spores prepared from lysozyme-treated sporulating cells of Bacillus subtilis AJ 1307. The enzyme was easily solubilized from the spores upon incubation at pH 10.5 at 37 degrees C. The transglutaminase activity was separated into two fractions upon purification by hydrophobic interaction chromatography (TG1 and TG2). Each enzyme was purified to electrophoretic homogeneity (about 1,000-fold). Both enzymes had the same molecular weight of 29,000 as estimated by SDS-PAGE, had the same N-terminal 30 amino acid sequence, and also showed the same optimal temperature (60 degrees C) and pH (8.2). The purified enzyme catalyzed formation of cross-linked epsilon-(gamma-glutamyl)lysine isopeptides, resulting in the gel-formation of protein solutions such as alphas-casein and BSA.
authors.affiliation: Central Research Laboratories, Ajinomoto Co., Inc., Kawasaki, Kanagawa, Japan. shunichi_suzuki@ajinomoto.com
authors.forename: S
authors.initials: S
authors.lastname: Suzuki
authors.forename: Y
authors.initials: Y
authors.lastname: Izawa
authors.forename: K
authors.initials: K
authors.lastname: Kobayashi
authors.forename: Y
authors.initials: Y
authors.lastname: Eto
authors.forename: S
authors.initials: S
authors.lastname: Yamanaka
authors.forename: K
authors.initials: K
authors.lastname: Kubota
authors.forename: K
authors.initials: K
authors.lastname: Yokozeki
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D020539:Sequence Analysis, Protein; D012995:Solubility; D013171:Spores, Bacterial; D013696:Temperature; D011503:Transglutaminases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D011503:Transglutaminases; D005033:Ethylmaleimide