• KSBB Journal
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• v.21 no.1 s.96
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• pp.20-27
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• 2006

The fibrinolytic activities of soluble proteins extracted from seeds of Coix lacryma-jobi L., Carthamus tinctorius L. and Malva venicillata L. were studied. Fibrinolytic activity of extract from C. lacryma-jobi L. showed 1.3 times higher than plasmin used as positive control. The fibrinolytic enzyme was confirmed and extracted directly from seed of C. lacryma-jobi L. by a fibrin zymography. The protein was composed of a single polypeptide and its apparent molecular weight was found to be 7.8 kDa, as judged by SDS-polyacrylamide gel electrophoresis. The effect of temperature for the proteolytic enzyme activity were stabilized above $50^{\circ}C$ and then dramatically decreased. Also, the enzyme activity was clearly inhibited by APMSF, PMSF and TPCK, suggesting that it is a member of the chymotrypsin-like serine pretense. In addition, effects of gamma-irradiated on seed of each plants were revealed that 8 Gy and 64 Gy were higher than others. This result shown that gamma-irradiation of seeds were capable to increase the fibrinolytic activity. All these results suggest the pretense is a fibrinolytic enzyme belong to a family of chymotrypsin-like serine pretense.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.224-224
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• 2004

Plasminogen activators (PAs)는 자궁분비액, 난포액, 정장물질 등을 포함한 여러 가지 세포외 분비액(extracellular fluids) 및 plasma에 풍부하게 존재하는 세포외 전구효소인 plasminogen을 plasmin으로 전환시키는 단백질분해효소이다. PAs는 섬유소용해, 배란, 착상 및 수정을 포함한 다양한 생리적인 과정에서 중요한 역할을 수행하는 것으로 알려져 있다. 따라서, 본 연구는 돼지의 체외수정 과정시 정자와 난자에서의 plasminogen activators activity의 변화를 SDS-PAGE와 zymography를 이용하여 검토하였다. (중략)

• Microbiology and Biotechnology Letters
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• v.23 no.1
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• pp.1-5
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• 1995

Bacterial strains showing the fibrinolytic activity were screened from Chungkook-jang and Natto (Japanese traditional soy food). The strains isolated from Natto revealed a high level of fibrinolytic activity, wherease nearly half of the isolates from Chungkook-jang did not show the relevant activity. However, one strain isolated from Chungkook-jang showed the highest fibrinolytic activity (1.84 plasmin unit), and subsequently identified as Bacillus species. The fibrinolytic strain was designated as Bacillus sp. CK 11-4.

• KSBB Journal
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• v.25 no.3
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• pp.265-270
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• 2010

Simvastatins are widely used to reduce endogenous cholesterol synthesis and improve hypercholesterolemia. Also, simvastatin have been shown to induce both angiogenic and angiostatic responses. In this study, It was attempted to resolve this controversy by studying the effects of simvastatin on the cell migration and invasion with the proteinases secretion and expression pattern. U-373-MG cells treated with low dose of simvastatin ($0.001{\sim}0.5\;{\mu}M$) showed the induction of migration and invasion compared with the addition of a control buffer. On the contrary, high dose of simvastatin ($1{\sim}20\;{\mu}M$) showed the reduction of migration and invasion compared with the addition of a control buffer. It was also showed that simvastatin-regulated migrative and invasive phenotypes were consistent with the secretion and expression pattern of matrix metalloproteinase-2 (MMP-2), MMP-9 and plasmin.

• Journal of Mushroom
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• v.13 no.1
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• pp.30-36
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• 2015

Our study investigated the fibrinolytic, thrombin inhibitory, anti-oxidative and anti-inflammatory activities of the water extract and solvent fractions isolated from Pleurotus ferulea. Fibrinolytic activity was investigated using the fibrin plate method. Thrombin inhibitory activity was used to analyze thrombin inhibitor assay. The DPPH assay was used to estimate anti-oxidative activity. Inhibition of NO production was measured for anti-inflammatory activity in LPS-activated murine RAW 264.7 macrophage cells. An MTS assay was used to evaluate the effects of the water extract and solvent fractions isolated from Pleurotus ferulea on cell viability. Our results showed the fibrinolytic activity to be strong in the ethyl acetate fraction at 1.33 plasmin units. The ethyl acetate fraction also showed high thrombin inhibitory activity at 94.45%. The anti-oxidative activity of the water extract was 37.01% and the anti-inflammatory activity of the chloroform fraction was 98.13%. These findings suggest that Pleurotus ferulea's extract and fractions could be applicable in the development of functional foods for the treatment and prevention of cardiovascular diseases.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.649-656
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• 2002

Fibrinolytic microorganisms were screened from 42 samples of Korean fermented food (7 kinds of Chungook-jang, 14 kinds of commercial Doen-Jang, 5 kinds of home-made Doen-jang, and 16 kinds of Jeot-gal), 15 samples of Japanese fermented food (5 kinds of home-made soybean paste, and 10 kinds of Natto), and 19 samples of Indonesian fermented food (Tempe) as well as starters of Meju (500 microflora from Korea, and 22 from China). Initially, 11 isolates with strong fibrinolytic activity were selected for further characterization. The fibrinolytic activity of the 11 isolates ranged from 89 to 199% of standard plasmin. Four strains, M5l from Korean fermented food (Meju), I 1-1, I 1-4, and I 5-1 from Indonesian fermented food (Tempe), were chosen based on the degree of activity and reproducibility, and identified as Staphylococcus sciuri, Citrobacter or Enterobacter, Enterococcus faecalis, and Bacillus subtilis, respectively. The first two isolates are pathogenic stains while the latter two are considered as GRAS (Generally Recognized As Safe). Fibrinolytic activity of E. faecalis, characterized and designated as BRCA-5, reached a maximum, when the producer was cultivated in Ml7 broth supplemented with 1.0% glucose for 5 h at 37$^{\circ}C$ with shaking at 180 rpm. Compared to commercial fibrinolytic enzymes, the cell-free culture supernatant of 5. faecaiis BRCA-5 showed stronger activity than plasmin and streptokinase, but similar degree of specific activity as nattokinase and urokinase, aud it also demonstrated anticoagulant and antiplatelet activity ex vivo. These features of E. faecalis make it an attractive agent as a biomaterial for health-promoting foods.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.7 no.3
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• pp.476-482
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• 2006

The aim of this work was to perform the screening and identification of the bacterium, MK-15 having the activity of fibrinolytic enzyme for the commercial use. Initially, strain MK-15 was enriched and isolated from naturally fermented soybean. Morphological and various physiological characteristics of the strain MK-15 was examined. The activity of fibrinolytic enzyme derived from supernatants of test culture MK-15 was performed by fibrin plate method for solid fibrinolytic activity. As the result, the fibrinolytic activity of MK-15 grown on the soybean media was about 2.7 times greater than that of plasmin used as stardard. 16S rRNA analyses revealed that strain MK-15 was 99.9% similar to Bacillus subtilis species cluster, and the bacterium was designated as Bacillus sp. MK-15. Strain MK-15 was registered in GenBank as [DQ163021].

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.271-276
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• 2003

Bacillus amyloliquefaciens D4, which produces a strongly fibrinolytic enzyme, was isolated from Chungkook-Jang, a traditional Korean soybean-fermented food. B. amyloliquefaciens D4 was mutated with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) to yield a series of mutants with increasing levels of fibrinolytic enzyme production. After mutation, a mutant D4-7 was obtained with fibrinolytic activity about eight times stronger than the parent strain. The fibrinolytic activity of B. amyloliquefaciens D4-7, reached a maximum, when the producer was cultivated in 2% Isolated Soy Protein (ISP) broth for 48 h at $37^{\circ}C$. Compared to commercial fibrinolytic enzymes, the cell-free culture supernatant of B. amyloliquefaciens D4-7 showed stronger activity than plasmin and streptokinase. The optimum temperature and pH were $50^{\circ}C$ and 10.0 and thermostability was increased by the addition of glycerol, glucose, and NaCl.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.97-104
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• 2008

Plasminogen activators (PAs) are serine protease that cleave plasminogen to form the active protease plasmin. PA/plasmin system playa role in mammalian fertilization and motility and acrosome reaction of sperm. The present study was undertaken to identify PAs in porcine gametes and investigate a possible role of plasminogen in in vitro fertilization in the pig. When boar spermatozoa were preincubated in a fertilization medium (mTBM) for 0, 2, 4 or 6 h, the activity of tPA-PAI ($110{\sim}117\;kDa$), tPA ($62{\sim}70\;kDa$), and uPA ($34{\sim}38\;kDa$) was observed in the sperm incubation medium and sperm sample. PA activities in the sperm incubation medium significantly (p<0.05) increased according to increasing incubation times, while PA activities in sperm significantly (p<0.05) decreased at the same times. In addition, the rate of acrosome reaction in spermatozoa increased by increasing culture times. When oocytes were separated from porcine cumulus-oocytes complexes at 0, 22 or 44 h of maturation culture, no PA activities were observed in cumulus free-oocyte just after aspiration from follicles. However, the activity of tPA-PAI ($108{\sim}113\;kDa$) and tPA ($75{\sim}83\;kDa$) was observed at 22 h of in vitro culture and significantly (p<0.05) increased as the duration of the culture increased. On the other hand, when porcine oocytes were activated by sperm penetration or calcium ionophore, plasminogen significantly (p<0.05) increased ZP dissolution time (sec) in activated oocytes by sperm penetration. These results suggest that supplementation of plasminogen to fertilization medium may playa positive role in the improvement of in vitro fertilization ability in the pig.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2090-2099
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• 2015

Recently, the cardiovascular disease has been widely problematic in humans probably due to fibrin formation via the unbalanced Western style diet. Although direct (human plasmin) and indirect methods (plasminogen activators) have been available, bacterial enzyme methods have been studied because of their cheap and mass production. To detect a novel bacterial fibrinolytic enzyme, 111 bacterial strains with fibrinolytic activity were selected from kimchi. Among them, 14 strains were selected because of their stronger activity than 0.02 U of plasmin. Their 16S rRNA sequence analysis revealed that they belong to Bacillus, Leuconostoc, Propionibacterium, Weissella, Staphylococcus, and Bifidobacterium. The strain B. subtilis ZA400, with the highest fibrinolytic activity, was selected and the gene encoding fibrinolytic enzyme (bsfA) was cloned and expressed in the E. coli overexpression system. The purified enzyme was analyzed with SDS-PAGE, western blot, and MALDI-TOF analyses, showing to be 28.4 kDa. Subsequently, the BsfA was characterized to be stable under various stress conditions such as temperature (4-40oC), metal ions (Mn²⁺, Ca²⁺, K²⁺, and Mg²⁺), and inhibitors (EDTA and SDS), suggesting that BsfA could be a good candidate for development of a novel fibrinolytic enzyme for thrombosis treatment and may even be useful as a new bacterial starter for manufacturing functional fermented foods.