title: Purification and characterization of an extracellular muramidase of Clostridium acetobutylicum ATCC 824 that acts on non-N-acetylated peptidoglycan.
abstract: An extracellular enzyme showing lytic activity on non-N-acetylated peptidoglycan has been isolated from Clostridium acetobutylicum ATCC 824. The lytic enzyme was purified to homogeneity by anion-exchange chromatography and gel filtration, with a recovery of 24%. The enzyme was monomeric and had an estimated molecular weight of 41,000 and an isoelectric point of 3.8. It has been characterized as a muramidase whose 23-amino-acid N terminus displayed 39% homology with the N,O-diacetyl muramidase of the fungus Chalaropsis sp. The muramidase hydrolyzed purified cell walls at an optimum pH of 3, with a maximum velocity of 9.1 mumol of reducing sugars released min-1 mg of muramidase-1 and a concentration of cell walls giving a half-maximum rate of 0.01 mg ml-1. Its activity was inhibited by glucosamine, N-acetylglucosamine, Hg2+, Fe3+, and Ag+ but not by choline. The muramidase-peptidoglycan complex rapidly dissociated before total hydrolysis of the chain and randomly reassociated on another peptidoglycan chain. The affinity of the muramidase was affected by the protein content and the acetylation of the cell wall.
authors.affiliation: Département de Génie Biochimique et Alimentaire, Institut National des Sciences Appliquées, Toulouse, France.
authors.forename: C
authors.initials: C
authors.lastname: Croux
authors.forename: B
authors.initials: B
authors.lastname: Canard
authors.forename: G
authors.initials: G
authors.lastname: Goma
authors.forename: P
authors.initials: P
authors.lastname: Soucaille
mesh_terms: D000107:Acetylation; D000438:Alcohols / Q000494:pharmacology; D000595:Amino Acid Sequence; D050260:Carbohydrate Metabolism; D003013:Clostridium / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D008969:Molecular Sequence Data; D009113:Muramidase / Q000302:isolation & purification / Q000378:metabolism*; D010457:Peptidoglycan / Q000378:metabolism*; D016415:Sequence Alignment; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D000438:Alcohols; D008670:Metals; D010457:Peptidoglycan; D009113:Muramidase