• The Korean Journal of Mycology
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• v.33 no.2
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• pp.69-74
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• 2005

Fibrinolytic enzyme has been isolated and purified from the edible mushroom, Lepista nuda. The apparent molecular mass of purified enzyme was estimated to be 34 KDa by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of the enzyme was Tyr-Pro-Ser-Pro-Ser-His-Gln-Thr-Ala-Val-Asn-Ala-Ile-Ile-X. It has a pH optimum at $7.0.{\sim}9.5$, suggesting that the purified enzyme is an alkaline protease. It shows the maximum fibrinolytic activity at $55^{\circ}C$. The fibrinolytic activity was inhibited by phenylmethylsulfonyl fluoride, indicating that the purified enzyme is a serine protease. The activity of the purified enzyme was totally inhibited by $Hg^{2+}$.

• Journal of Life Science
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• v.12 no.3
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• pp.357-362
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• 2002

A bacterial strain showing the fibrinolytic activity was screened from korean traditional soybean products. For the identification, the strain was investigated morphology and biochemical characteristics and it was classified to Bacillus subtilis. The strain had high fibrinolytic activity in Chungkukjang. The optimum fermentation condition of temperature and time were 37$^{\circ}C$ and 24hour. The pH in Chungkukjang was gradually alkalized during fermentation. The fibrinolytic enzyme in Chungkukjang stable at heft treatment; After heating at 6$0^{\circ}C$ and 8$0^{\circ}C$ for 30 min, the fibrinolytic activity remained 75% and 40%, respectively.

• Biomedical Science Letters
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• v.17 no.2
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• pp.157-165
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• 2011

In this study, we investigated the biological activities of Carthamus tinctorius including antioxidative, fibrinolytic, thrombin inhibitory, ${\alpha}$-glucosidase inhibitory and collagenase inhibitory activities. Carthamus tinctorius, hot water extract was fractionated into hexane, chloroform, ethyl acetate, butanol and water fraction. Each of these was assayed individually. The hot water extract showed high antioxidative activity and thrombin inhibitory activity at 90.17% and 97.10% respectively. In the fraction activity tests, chloroform fraction showed the highest antioxidative activity at 81.85%. The fibrinolytic activity was strong only in the butanol fraction at 0.70 plasmin units/ml. The thrombin inhibitory activities of hexane, ethyl acetate and butanol fractions were 97.35%, 86.74% and 93.18% respectively. In collagenase inhibitory activity test, hexane fraction showed the highest activity at 87.78%. In conclusion, the hot water extract and solvent fractions of Carthamus tinctorius L can be used as a material for the development of biofunctional tea and foods respectively.

• 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.

• Korean Journal of Food Science and Technology
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• v.31 no.2
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• pp.553-557
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• 1999

Five fungi (mushrooms), Daedaleopsis styracina, Trichaptum abietium, Coriolus versicolor, Pisolithus tinctorius and Tricholomopsis decora, were screened and examined the fibrinolytic activity and specificity. The extracts of mushrooms showed a level of fibrinolytic activity that was about 3-4 times higher than that of plasmin 1.0 unit. In particular, Pisolithus tinctorius of them showed the greatest enzyme activity (4.71 plasmin unit/mL) by fibrin plate assay, and the highest specificity (1.32 plasmin unit/mL) using chromogenic substrate (N-p-Tosyl-Gly-Pro-Lys p-nitroanilide) by Tricholomopsis decora. And the same molecular mass 54 and 61kDa showing the fibrinolytic activity obtained from all fruiting bodies were confirmed, and it was found that Trichaptum abietium and Tricholomopsis decora have a strong fibrinolytic enzyme with an apparent size of 100 kDa and 84 kDa, respectively on SDS-fibrin zymography activity assay.

• 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.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.87-90
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• 1998

Various bacterial strains that secret extracellular fibrinolytic enzyme were screened from Doen-Jang, a traditional soybean fermented food in Korea. Five microbes of them were identified to be Bacillus sp. strains according to Bergey's manual of systematic bacteriology. The culture filtrates of B. amyloliquefaciens (2.46 plasmin unit/ml) and B. pantothenticus (3.82 plasmin unit/ml) showed a level of fibrinolytic activity that was about three times higher than that of plasmin 1.0 unit and Bacillus subtilis showed the highest fibrinolytic activity (4.94 plasmin unit/ml). All of the extracellular proteases showing the fibrinolytic activity are confirmed by SDS-PAGE followed by reverse fibrin zymogram activity assay and we proposed that some of the fibrinolytic enzymes from this work are novel enzymes.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.522-529
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• 2019

A Bacillus strain, SJ4, exhibiting strong fibrinolytic activity was isolated from saeu (shrimp, Acetes chinensis) jeotgal, a Korean traditional fermented food and was identified as B. subtilis. The B. subtilis SJ4 strain can grow at a NaCl concentration of up to 15% (w/v). The fibrinolytic activity of B. subtilis SJ4 (152.0 U/ml) cultured in Luria-Bertani (LB) broth for 48 h at 37℃ with aeration was higher than that of B. subtilis SJ4 cultured in TSB (124.5 U/ml) under same culture conditions. The major proteins in the LB culture supernatant of B. subtilis SJ4 were analyzed by SDS-PAGE, which revealed three major bands (23, 25, and 28 kDa). The band (23 kDa) with strong fibrinolytic activity, analyzed on fibrin zymogram, was observed at 60-96 h of cultivation. The aprESJ4 gene encoding the major fibrinolytic enzyme, AprESJ4, was cloned by PCR. The aprESJ4 gene sequence exhibited high similarities with the fibrinolytic gene sequences of other Bacillus species. The amino acid sequence of AprESJ4 exhibited 98.9 and 98.4% similarity with subtilisin NAT and AprE2 of B. subtilis, respectively. Hence, B. subtilis SJ4 can be a potential starter culture for jeotgal products.

• 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.

• Biomedical Science Letters
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• v.11 no.1
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• pp.63-69
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• 2005

We investigated the changes of fibrinolytic and antioxidative activities of wild or cultivated Codonopsis lanceolata, which were packed in woven polypropylene (WP) film or low density polyethylene (LDPE) film and stored for 0, 15, and 30 days at refrigerated $(2\~4^{\circ}C)$ or room $(l8\~20^{\circ}C)$ temperature (So we have 16 samples, and wrote them as the abbreviated words such as W-WP-RE-15, W-WP-RO-15, W-LDPE-RE-15, W-LDPE-RO-15, W-WP-RE-30, W-WP-RO-30, W-LDPE-RE-30, W-LDPE-RO-30, C-WP-RE-15, C-WP-RO-15, C-LDPE-RE-15, C-LDPE-RO-15, C-WP-RE-30, C-WP-RO-30, C-LDPE-RE-30, C-LDPE-RO-30). Fibrinolytic activity of fresh Codonopsis lanceolata cultivated in the mountain or field were 0.8 unit (plasmin unit/ml) or 7.3 units, respectively. In descending order, the activities of wild Codonopsis lanceolata stored for 15 days were as followed; LDPE-RE (0.70 unit), WP-RO (0.52 unit), WP-RE (0.45 unit), and LDPE-RO (0.30 unit). After 30 days, fibrinolytic activities of them decreased to 0.47 unit (LDPE-RE), 0.28 unit (WP-RO), 0.21 unit (WP-RE), and 0.30 unit (LDPE-RO). Considering from the point of fibrinolytic activity, the optimal storage condition of wild Codonopsis lanceolata was packing with LDPE film and storing at $4^{\circ}C$. The change of fibrinolytic activities of Codonopsis lanceolata cultivated in the field revealed the similar trend as wild samples, but the fibrinolytic activities of 30 days-stored samples were maintained better than the wild Codonopsis lanceolata. Fibinolytic activity of wild Codonopsis lancealata was increased by heating for 5 min at $100^{\circ}C$ and decreased by addition of NaCl. Antioxidative activities of Codonopsis lanceolata were also compared from the electron donating activity. Fresh Codonopsis lanceolata had about $70\%$ of electron donating activity. Independent of cultivation area, electron donating activity dropped to $19\~74\%$ (wild Codonopsis lanceolata) and $27\~59\%$ (cultivated Codonopsis lanceolata) during 15 days storage. But after 30 days storage, we obtained the unexpected results, which meant that the activities were higher activities than 15 days-stored samples or even though the fresh samples. In general, Codonopsis lanceolata could maintain antioxidative activities most strongly with LDPE film and chilled condition.