title: Fibrinolysis and anticoagulant potential of a metallo protease produced by  Bacillus subtilis K42.
abstract: In this study, a potent fibrinolytic enzyme-producing bacterium was isolated from soybean flour and identified as Bacillus subtilis K42 and assayed in vitro for its thrombolytic potential. The molecular weight of the purified enzyme was 20.5 kDa and purification increased its specific activity 390-fold with a recovery of 14%. Maximal activity was attained at a temperature of 40 degree C (stable up to 65 degree C) and pH of 9.4 (range: 6.5 - 10.5). The enzyme retained up to 80% of its original activity after pre-incubation for a month at 4 degree C with organic solvents such as diethyl ether  (DE), toluene (TO), acetonitrile (AN), butanol (BU), ethyl acetate (EA), ethanol (ET), acetone (AC), methanol (ME), isopropanol (IP), diisopropyl fluorophosphate (DFP), tosyl-lysyl- chloromethylketose (TLCK), tosyl-phenylalanyl chloromethylketose (TPCK), phenylmethylsulfonylfluoride (PMSF) and soybean trypsin inhibitor (SBTI). Aprotinin had little effect on this activity. The presence of ethylene diaminetetraacetic acid (EDTA), a metal-chelating agent and two metallo protease inhibitors, 2,2'-bipyridine and o-phenanthroline, repressed the enzymatic activity significantly. This, however, could be restored by adding Co2+ to the medium. The clotting time of human blood serum in the presence of this enzyme reached a relative PTT of 241.7% with a 3.4-fold increase, suggesting that this enzyme could be an effective antithrombotic agent.
authors: Department of Microbiology, Faculty of Science, Zagazig University, Zagazig, Egypt 44519.
authors: Wesam A
authors: WA
authors: Hassanein
authors: Essam
authors: E
authors: Kotb
authors: Nadia M
authors: NM
authors: Awny
authors: Yehia A
authors: YA
authors: El-Zawahry
mesh_terms: D000925:Anticoagulants / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D001412:Bacillus subtilis / Q000201:enzymology* / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D057075:Enzyme Assays; D004795:Enzyme Stability; D005342:Fibrinolysis / Q000187:drug effects*; D006801:Humans; D045726:Metalloproteases / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D019216:Metals, Heavy / Q000737:chemistry; D011480:Protease Inhibitors / Q000737:chemistry; D013025:Glycine max / Q000382:microbiology
chemical_list: D000925:Anticoagulants; D001426:Bacterial Proteins; D019216:Metals, Heavy; D011480:Protease Inhibitors; D045726:Metalloproteases