title: Purification and characterization of a novel fibrinolytic enzyme from Bacillus sp. nov. SK006 isolated from an Asian traditional fermented shrimp paste.
abstract: Bacillus sp. nov. SK006 producing four extracellular fibrinolytic enzymes was isolated from fermented shrimp paste, a traditional and popular Asian seasoning. One fibrinolytic enzyme was purified to homogeneity with a molecular mass of 43-46 kDa by SDS-PAGE and gel filtration chromatography. The specific activity was determined to be 11.2 units/mg using plasmin as a standard. The enzyme displayed optimal activity at 30 degrees C and pH 7.2. It was stable below 40 degrees C for 4 h between pH 5.0 and pH 11.0. Zinc ion stimulated the enzyme activity whereas Cu2+, Ca2+, Fe3+, and Hg2+ caused its inhibition. The fibrinolytic activity was strongly inhibited by PMSF and moderately inhibited by EDTA as well as PCMB. The enzyme exhibited a higher affinity toward N-Succ-Ala-Ala-Pro-Phe-pNA and was able to degrade fibrin clots either by forming active plasmin from plasminogen or by direct fibrinolysis. The N-terminal amino acid sequence was found to be AQSVPYEQPHLSQ, which is different from that of other known fibrinolytic enzymes.
authors.affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Jiangsu, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Hua
authors.forename: Bo
authors.initials: B
authors.lastname: Jiang
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Mine
authors.forename: Wanmeng
authors.initials: W
authors.lastname: Mu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000201:enzymology*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors; D005285:Fermentation; D005341:Fibrinolysin / Q000378:metabolism; D005342:Fibrinolysis; D005421:Flavoring Agents / Q000737:chemistry; D005516:Food Microbiology; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D033561:Penaeidae; D010958:Plasminogen / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D004791:Enzyme Inhibitors; D005421:Flavoring Agents; D010958:Plasminogen; D010450:Endopeptidases; D005341:Fibrinolysin