• Microbiology and Biotechnology Letters
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• v.39 no.4
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• pp.330-336
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• 2011

Bacterial strains exhibiting fibrinolytic activity were screened from traditional Korean soybean sauce. The Fibrinolytic activities of the various isolated microorganism were further examined and the superior strain YJ11-21 was selected for further analyses. Gene sequence analysis of 16S rDNA of the YJ11-21 strain revealed Bacillus licheniformis. Optimal culture conditions were investigated in order to maximize the production of the fibrinolytic enzyme by YJ11-21. Amongst the carbon sources tested, glucose was the most effective for enzyme production and amongst the nitrogen sources tested, yeast extract was seen to be the most effective. A one percent addition of NaCl to the medium resulted in the highest fibrinolytic activity. Interestingly, a 10% addition of NaCl resulted in a high activity together with a high cell growth rate. Therefore, YJ11-21 is speculated of being a halotolerant. The optimum pH and temperature for enzyme production were a pH of 9.0 and $30^{\circ}C$, respectively.

• Journal of Life Science
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• v.15 no.4 s.71
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• pp.513-521
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• 2005

A bacterium producing five fibrinolytic isozymes was isolated from black bean chung kuk. The bacterium was identified as Bacillus subtilis BB-1 by 16s rDNA sequence homology search. A gene out of five fibrinolytic genes in the Bacillus subtilis BB-1 was cloned by shot-gun method. A Cla I DNA fragment of B. subtilis BB-1 chromosome was cloned in to pBluescript II SK(-) and showed the fibrinolytic activity to bacterial cells. The Cla I DNA fragment was sequenced and the sequences did not show homology with gene for protease or fibrinolytic enzyme genes in other organisms. The Cla I DNA fragment was reduced to 2,142 bp by activity-guided PCR cloning method. The optimum pH and temperature of the enzyme were 5.0 and $35^{\circ}C$, respectively. Substrate specificity of the fibrinolytic enzyme was detected in skim milk, casein, gelatin and blood agar plates. The activity of the enzyme was not detected with these substrates. Taken together, this enzyme is a new fibrinolytic enzyme and may be used to prevent thrombosis and arteriosclerosis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.2
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• pp.439-442
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• 2004

Bacterial strains showing the firinolytic and $\beta$-glucosidase activity were screened from Doeniang. The strain of KH-15 revealed a high level of fibrinolytic and $\beta$-gluocosidase activity. The isolated bacterium was identified and desingnated as Bacillus sp. KH-15. The carbon, nitrogen and salts sgnificantly influenced te fibrinolytic enzyme and $\beta$-glucosidase production. The optimized composition of medium appeared to be 2％ glucose, 0.5% yeast extract and 0.1% calcium chloride. The optimum pH and temperature for fibrinolytic enzyme and $\beta$-glucosidase activities were pH 7∼8, 4$0^{\circ}C$ and pH 6∼8, 30∼4$0^{\circ}C$, respectively.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.370-374
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• 2010

A gene encoding the major secreted fibrinolytic protein of Bacillus amyloliquefaciens CH86-1 was cloned from genomic DNAs. DNA sequencing showed that the gene, aprE86-1, could direct the synthesis of a mature protein 275 amino acids in length after processing. When aprE86-1 was introduced into B. subtilis, a mature 27-kDa protein was produced as expected. The fibrinolytic activity of the B. subtilis transformant (TF) was higher than that of B. amyloliquefaciens CH86-1, showing the possibility of increasing the fibrinolytic activity of Bacillus strains through genetic engineering.

• Preventive Nutrition and Food Science
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• v.19 no.3
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• pp.213-219
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• 2014

Bacillus pumilus 2.g isolated from gembus, an Indonesian fermented soybean cake, secretes several proteases that have strong fibrinolytic activities. A fibrinolytic enzyme with an apparent molecular weight of 20 kDa was purified from the culture supernatant of B. pumilus 2.g by sequential application of ammonium sulfate precipitation, ion-exchange chromatography, and hydrophobic chromatography. The partially purified enzyme was stable between pH 5 and pH 9 and temperature of less than $60^{\circ}C$. Fibrinolytic activity was increased by 5 mM $MgCl_2$ and 5 mM $CaCl_2$ but inhibited by 1 mM phenylmethylsulfonyl fluoride (PMSF), 1 mM sodium dodecyl sulfate (SDS), and 1 mM ethylenediaminetetraacetic acid (EDTA). The partially purified enzyme quickly degraded the ${\alpha}$ and ${\beta}$ chains of fibrinogen but was unable to degrade the ${\gamma}$ chain.

• Preventive Nutrition and Food Science
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• v.11 no.2
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• pp.127-132
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• 2006

Bacillus sp. strain K-l, which produces a strong fibrinolytic enzyme, was isolated from chongkukjang, a traditional Korean fermented soybean paste. The fibrinolytic enzyme was purified from chongkukjang base by using ammonium sulfate fractionation and chromatographic techniques. Purified enzyme, CK K-1 was demonstrated to be homogeneous by SDS-PAGE and isoelectric focusing electrophoresis, and has molecular mass of a 12.4 kDa and a pI of 8.0. The optimal reaction pH value and temperature were 8.0 and $40^{\circ}C$, respectively. Phenyl-methyl-sulfonyl-fluoride (PMSF; serine protease inhibitor), ethylene-diamine-tetra-acetic acid (EDTA; metallo protease inhibitor), copper ion, ferric ion and lead ion inhibited the enzyme activity. These results indicated that the fibrinolytic enzyme is a metallo-serine protease and different from nattokinase and chongkukjangkinase.

• The Korean Journal of Mycology
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• v.31 no.2
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• pp.114-116
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• 2003

Platelet aggregation inhibitory and fibrinolytic activities of water and ethanol extracts from mushrooms were studied. The highest platelet aggregation inhibitory activity was 81.2% in the ethanol extract from fruiting body of Inonotus obliquus ASI 74006 and also were high in the ethanol extract from fruiting bodies of Fomitella fraxinea. The ethanol extract from the mycelia of Agaricus blazei Murill. ASI 1174 showed the strongest fibrinolytic activity as 9.6 unit. However fibrinolytic activities of other mushrooms were low or negligible.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.7
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• pp.1066-1070
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• 2003

Effects of cereal kojis and legumes on alcohol fermentation and fibrinolytic activities of Korean traditional wines were investigated. The Korean traditional wine which was brewed by addition of 10% barley koji into the mash showed the greatest fibrinolytic activity of 20.0 U and good ethanol productivity (16.8%). The fibrinolytic activity was increased up to 26.0 U by addition of 50% of mungbean and its acceptability were improved by addition of jujube (3%) into the mash.

• Natural Product Sciences
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• v.8 no.2
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• pp.66-70
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• 2002

The in vitro anticoagulant and fibrinolytic activities of crude extracts from insects were evaluated in order to find effective therapeutic drugs for the treatment of myocardial and cerebral thrombosis. We prepared three types of extracts (water, methanol and ethylacetate) from 28 insects for use as raw materials for the activity assays. The fibrinolytic activity was tested using the fibrin plate method and the activated partial thromboplastin time and thrombin time were measured for blood clotting activity. With regards to the fibrinolytic system, water extracts of six kinds of insects displayed a remarkable level of activity with a plasmin-like action. The water extracts of [Catharsius molossus, Eupolyphaga sinensis, Huechys sanguinea, Mantidis $o\ddot{o}theca$, Mimela splendens, and Polistes mandarinus (Vespae Nidus)] exhibited the activity. On the other hand, the methanol extracts did not display any fibrinolytic activity. In terms of the coagulation system, an aqueous extract of silkworm Tongchunghacho (Paecilomyces japonica), Oxya japonica japonica and Buthus martensi (Scorpion) increased the clotting time significantly longer (181 times) than the control. These results suggest that crude drugs from insects are useful sources for the development of new drugs for use in treatments involving blood coagulation and fibrinolysis.

• Journal of Life Science
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• v.8 no.6
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• pp.737-740
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• 1998

Effect of oral administration with fibrinolytic enzyme isolated from fermented anchovy(the traditional fermented food in Korea called Myulchi Jeot-gal) and its functionally active enzyme to rat, activation of plasma fibrinolysis was observed. The euglobulin fibrinolytic activities and the plasma levels of H-D-Val-Leu-Lys-pNA(S-2251) amidolysis reached a maximum at 3 hours after the administration to rat. And euglobulin Iysis time(ELT) value after oral admi-nistration showed its activity 2∼3 hours later. From the above result, it was confirmed the enzyme activity in blood by oral administration fibrinolytic enzyme through animal experiment.