• Preventive Nutrition and Food Science
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• v.14 no.1
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• pp.90-93
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• 2009

Cheonggukjang was prepared from soybean inoculated with B. licheniformis ATCC 10716 cells transformed with pHY3-5 carrying a fibrinolytic enzyme gene. During the 54 hr of fermentation at $37^{\circ}C$, fibrinolytic activities of cheonggukjang were significantly higher than cheonggukjang fermented with B. licheniformis 10716 control cells. The plasmid, pHY3-5 was stably maintained during the 54 hr without antibiotic selection and more than 52% of cells retained the plasmid.

• Journal of Microbiology and Biotechnology
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• v.21 no.5
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• pp.515-518
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• 2011

A gene encoding bacillopeptidase F, bpr86-1, was cloned from B. amyloliquefaciens CH86-1 isolated from cheonggukjang. This gene could encode a preproenzyme of 1,431 amino acids. When bpr86-1 was introduced into B. subtilis WB600 via pHY300PLK, an E. coli-Bacillus shuttle vector, the transformant showed fibrinolytic activity. During growth on LB, the fibrinolytic activity of cells increased sharply when they entered the stationary phase. The highest activity (761.4 mU/mg protein) was observed at 96 h of cultivation.

• The Korean Journal of Mycology
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• v.26 no.4 s.87
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• pp.589-593
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• 1998

Extracts from 65 species of mushrooms in mount Chiak were screened for their fibrinolytic activities. Extracts from Armillariella mellea, Calocybe sp., Lepista nuda and Trichaptum abietinum showed to have almost twice of activity of plasmin 1.5 U/ml, 193%, 213%, 198%, and 193% respectively. Collybia maculata showed 98% of activity, and Coprinus comatus and Lepiota sp. 56% and 58% activities, respectively. Other two, Agrocybe sp. and Stropharia rugosoannulata were less than 10% of activities.

• Proceedings of the KOSOMBE Conference
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• v.1998 no.11
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• pp.227-228
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• 1998

Lumbrokinase, potent fibrinolytic enzyme purified from earthworm, was immobilized onto polyurethane valves using photoreaction, photoreactive polyallyl-amino as a photoreactive linker. For evaluation of blood compatibility, lumbrokinase immobilized polymer valves were assembled into the total artificial heart (TAH). This TAH was implanted to 60kg healthy lamb for 1-3 days with the cardiac output 5 L/min. In the control lamb, the valves were untreated, in ore other, only valves on the right were treated, and in the remaining animal, only those on the left. To facilitate the thrombus formation, low doses of heparin were administered. For evaluation of the immobilized lumbrokinase, thrombus formation, proteolytic and fibrinolytic activity was measured. This data shows that lumbrokinase-treated polyurethane valves lead to decreased thrombus formation in vivo, and that their biocompatibility is therefore higher than that of untreated valves.

• Journal of Biomedical Engineering Research
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• v.26 no.3
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• pp.157-161
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• 2005

Direct injection of a fibrinolytic agent to the intraarterial thrombosis may increase the effectiveness of thrombolysis by enhancing the permeation of thrombolytic agents into the blood clot. Permeation of fibrinolytic agents into a clot is influenced by the surface pressure, which is determined by the injection velocity of fibrinolytic agents. In order to calculate the pressure distribution on the clot surface for different jet velocities (1, 3, 5 m/sec) and nozzle arrangements (1, 9, 17 nozzles), computational fluid dynamic methods were used. Thrombolysis of a clot was mathematically modeled based on the pressure and lysis front velocity relationship. Direct injection of a thrombolytic agent increased the speed of thrombolysis significantly and the effectiveness was increased as the ejecting velocity increased. The nine nozzles model showed about $20\%$ increase of the lysed volume, and the one and seventeen nozzles models did not show significant differences. The wall shear stress decreased as the number of nozzles increased, and the wall shear stress in most vessel wall was lower than 25 Pa. The results implied that thrombolysis could be accelerated by direct injection of a drug with the moderate velocity without damaging the blood vessel wall.

• Tuberculosis and Respiratory Diseases
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• v.76 no.3
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• pp.127-130
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• 2014

The risk of dying from a pulmonary embolism (PE) is estimated to be about 30% if inotropic support is required and no cardiopulmonary arrest occurs. Fibrinolysis in massive PE is regarded as a life-saving intervention, unless there is a high risk of bleeding following the use of the fibrinolytic therapy. Rivaroxaban is an oral factor Xa inhibitor, however its anticoagulation effects before or after administration of fibrinolytics in massive PE are still unknown. Two patents were admitted: 61-year-old woman with repeated syncope, and a 73-year-old woman was admitted with dyspnea and poor oral intake. Systemic arterial hypotension with radiologic confirmation led to a diagnosis of massive PE in both patients. Rivaroxaban was administered before in one, and after firbrinolytic therapy in the other. One showed similar efficacy of rivaroxaban with currently used anticoagulants after successful fibrinolysis, and the other one without antecedent administration of the fibrinolytic agent showed unfavorable efficacy of rivaroxaban.

• Archives of Pharmacal Research
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• v.21 no.4
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• pp.374-377
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• 1998

The freeze-dried powder of Lumbricus rubellus earthworm was administered orally to rats and its fibrinolytic and antithrombotic effects were investigated. The fibrinolytic activity of plasma was determined by measuring the plasmin activity of the euglobulin fraction and was increased to two-folds of the control at a dose of 0.5 g/kg/day and five times with 1 g/kg/day after 4-day administration. The antithrombotic effect was studied in an arterio-venous shunt model of rats. The thrombus weight decreased significantly from 43.2 mg to 32.4 mg at a dose of 0.5 g/kg/day after 8-day treatment. The level of fibrinogen/fibrin degradation product (FDP) in serum was elevated in a dose-dependent manner during the treatment period. On the 8th day after administration, the FDP value was increased to 7.7 $\mu\textrm{g}$g/ml compared with the control value of 3.3 $\mu\textrm{g}$g/ml. These results support that earthworm powder is valuable for the prevention and/or treatment of thrombotic conditions.

• Journal of Microbiology
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• v.44 no.6
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• pp.622-631
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• 2006

In this study we purified a fibrinolytic enzyme from Cordyceps militaris using a combination of ion-exchange chromatography on a DEAE Sephadex A-50 column, gel filtration chromatography on a Sephadex G-75 column, and FPLC on a HiLoad 16/60 Superdex 75 column. This purification protocol resulted in a 191.8-fold purification of the enzyme and a final yield of 12.9 %. The molecular mass of the purified enzyme was estimated to be 52 kDa by SDS-PAGE, fibrin-zymography, and gel filtration chromatography. The first 19 amino acid residues of the N-terminal sequence were ALTTQSNV THGLATISLRQ, which is similar to the subtilisin-like serine protease PR1J from Metarhizium anisopliae var. anisopliase. This enzyme is a neutral protease with an optimal reaction pH and temperature of 7.4 and $37^{\circ}C$, respectively. Results for the fibrinolysis pattern showed that the enzyme rapidly hydrolyzed the fibrin $\alpha$-chain followed by the $\gamma$-$\gamma$ chains. It also hydrolyzed the $\beta$-chain, but more slowly. The A$\alpha$, B$\beta$, and $\gamma$ chains of fibrinogen were also cleaved very rapidly. We found that enzyme activity was inhibited by $Cu^{2+}$ and $Co^{2+}$, but enhanced by the additions of $Ca^{2+}$ and $Mg^{2+}$ ions. Furthermore, fibrinolytic enzyme activity was potently inhibited by PMSF and APMSF. This enzyme exhibited a high specificity for the chymotrypsin substrate S-2586 indicating it's a chymotrypsin-like serine protease. The data we present suggest that the fibrinolytic enzyme derived from the edible and medicinal mushroom Cordyceps militaris has fibrin binding activity, which allows for the local activation of the fibrin degradation pathway.

• Korean Journal of Food Science and Technology
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• v.44 no.5
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• pp.600-606
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• 2012

A Bacillus sp. that produces fibrinolytic enzyme was isolated from Cheonggukjang, a traditional Korean soybean-fermented food. According to 16S rRNA gene base sequencing, the bacillus was identified as a variety of Bacillus subtilis, and named Bacillus subtilis IDCC 9204. Fibrinolytic enzyme NK-IL9204 was stable up to $60^{\circ}C$ and within pH range of 5-10. Purified NK-IL9204 was detected through fibrin zymography. The molecular weight and isoelectric point of the enzyme were estimated to be 27.7 kDa and 6.7 by SDS-PAGE and 2D electrophoresis, respectively. Its amino acid sequence was similar to that of nattokinase (identities 99.5%) and different from that of nattokinase BPN (identities 86.4%). The plasma fibrinolytic activity of NK-IL9204 was measured by euglobulin clot lysis times (ECLT). The NK-IL9204 was orally administered to SD rats for 3 weeks (1,000 FU/rat/day). The ECLT was significantly shortened by supplementation of NK-IL9204.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.299-305
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• 2006

In this study, we have isolated and identified a proteolytic bacterium from conventional Cheongkukjang. We also characterized several bioactivity of Cheongkukjang, which was fermented by an isolated strain. One strain out of about $10^4$ strains obtained from Cheongkukjang showed relatively high proteolytic activity was selected and named as a Bacillus subtilis LSH805 strain. White soy-bean Cheongkukjang possessed less odor and more viscous substance than black soy-bean Cheongkukjang. Cheongkukjang showed fibrinolytic activity, and about 1,500 mg fibrin was degraded after 20 h incubation. Although nitric oxide (NO) assays of soy-bean and Cheongkukjang were almost the same, their activities were significantly higher than that of no treatment. Activity of water fraction of Cheongkukjang was somewhat higher than that of soy-bean. Fibrinolytic and NO assays of Cheongkukjang suggest that Cheongkukjang, which was fermented by an isolated strain may be a useful candidate for natural fibrinolytic and macrophage-stimulating agents.