• 생명과학회지
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• 제9권4호
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• pp.489-492
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• 1999

A bacteial strain which can produce the extracellular fibrinolytic enzyme was isolated from Jeot-Gal (anchovy) that was Korean traditional salt-fermented fish. The isolated bacterium was identified to be a strain of Bacillus sp. The optimal medium for fibrinolytic enzyme production was determined to consist of 5 g maltose, 10 g defatted soybean, 20 g sodium chloride, 1.75 g K2HPO4 per liter (pH 7.0)

• 대한의생명과학회지
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• 제8권3호
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• pp.173-177
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• 2002

To investigate the fibrinolytic activities of the Korean basidiomycetes, the Extracts of 50 wild mushrooms were tested for their fibrinolytic activites. Extract from Tricholoma sejunctum showed 175% increased activity to that of plasmin 1.0 U/ml. Marasmius siccus showed 54% of activity, and Laetiporus sulphureus var. miniatus and Macrolepiota procera, 43% and 26% activities, respectively to that of plasmin. But, Cystoderma amianthinum, Lepiota sp., Coprinus sp., Lycoperdon sp. were less than 10% of plasmin activity.

• Journal of Korean Neurosurgical Society
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• 제66권3호
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• pp.263-273
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• 2023

While the survival rate of preterm infants has increased dramatically over the last few decades, intraventricular hemorrhage and subsequent hydrocephalus remain major unsolved problems in neonatal intensive care. Once intraventricular hemorrhage occurs, severe neurological sequelae are inevitable. Treatment of this complicated pathology and achievement of favorable neurofunctional outcomes in fragile infants are crucial challenges for pediatric neurosurgeons. Fibrinolytic therapy, which chemically dissolves hematoma, is a promising and useful treatment method. In this paper, the historical background of fibrinolytic therapy for post-intraventricular hemorrhagic hydrocephalus in preterm infants is reviewed and a recent method of fibrinolytic therapy using urokinase is introduced.

• 한국미생물·생명공학회지
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• 제26권6호
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• pp.507-514
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• 1998

국내 전통식품인 청국장으로부터 강력한 fibrinolytic activity를 갖는 K-54주를 분리, 동정한 결과 B. subtilis로 확인되었으며 이 균으로부터 강력한 갖는 효소를 정제하였다. 정제도는 약 66.9배 증가하였으며 약 10.1%의 수율을 보였다. B. subtilis K-54에서 분리한 fibrinolytic enzyme의 최적 활성 pH가 8-12사이였으며 50-75$^{\circ}C$사이에서 최적 활성을 보였다. 정제된 효소의 분자량은 약 29 kDa이었고, 등전점은 8.67로 나타났으며 Vmax값은 47.16%, Km값은 0.31(3.1 mg/$m\ell$)였다. N-말단 아미노산 서열 분석 결과 B. subtilis K-54주에서 정제한 fibrinolytic enzyme는 Ala-Gly-Ser-Val-Pro-Try-Gly-Ser이었으며 serine protease저해제인 PMSF에 강력한 활성 저해를 보여 serine protease계열의 효소임을 확인할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제29권3호
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• pp.347-356
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• 2019

Bacillus sp. BS2 showing strong fibrinolytic activity was isolated from sea squirt (munggae) jeotgal, a traditional Korean fermented seafood. BS2 was identified as B. velezensis by molecular biological methods. B. velezensis BS2 grows well at 15% NaCl and at $10^{\circ}C$. When B. velezensis BS2 was cultivated in TSB broth for 96 h at $37^{\circ}C$, the culture showed the highest fibrinolytic activity ($131.15mU/{\mu}l$) at 96 h. Three bands of 27, 35 and 60 kDa were observed from culture supernatant by SDS-PAGE, and fibrin zymography showed that the major fibrinolytic protein was the 27 kDa band. The gene (aprEBS2) encoding the major fibrinolytic protein was cloned, and overexpressed in heterologous hosts, B. subtilis WB600 and E. coli BL21 (DE3). B. subtilis transformant showed 1.5-fold higher fibrinolytic activity than B. velezensis BS2. Overproduced AprEBS2 in E. coli was purified by affinity chromatography. The optimum pH and temperature were pH 8.0 and $37^{\circ}C$, respectively. $K_m$ and $V_{max}$ were 0.15 mM and $39.68{\mu}M/l/min$, respectively, when N-succinyl-Ala-Ala-Pro-Phe-pNA was used as the substrate. AprEBS2 has strong ${\alpha}$-fibrinogenase and moderate ${\beta}$-fibrinogenase activity. Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at $10^{\circ}C$, B. velezensis BS2 can be used as a starter for jeotgal.

• 한국미생물·생명공학회지
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• 제48권4호
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• pp.439-446
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• 2020

Two Bacillus strains, K3 and K208, both demonstrating strong fibrinolytic activities were isolated from Kimchi, a traditional Korean preparation of fermented vegetables. Isolates were subjected to various molecular biology based identification methods including RAPD-PCR and identified as B. subtilis and B. velezensis, respectively. Tryptic soy broth (TSB) was found to best maintain both the growth and the fibrinolytic activity of these strains. Culture supernatants were analyzed by SDS-PAGE and fibrin zymography, and the results indicate that a 40 and 27 kDa band seem to be responsible for the fibrinolytic activities of these two isolates and the 27 kDa band was subsequently identified as the mature form of AprE, the major fibrinolytic enzyme. Thus the aprE genes were cloned and the translated amino acid sequences demonstrated 99.3% identity with each other, and 86.5% identity with BsfA, a fibrinolytic enzyme from B. subtilis ZA400 also isolated from Kimchi, and AprE2, a fibrinolytic enzyme from B. subtilis CH3-5 isolated from Cheonggukjang, a traditional Korean fermented soy. Given this B. subtilis K3 and B. velezensis K208 may be promising starter cultures in the production of fermented foods.

• Journal of Microbiology and Biotechnology
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• 제31권6호
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• pp.833-839
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• 2021

Bacillus subtilis CH3-5 isolated from cheonggukjang secretes a 28 kDa protease with a strong fibrinolytic activity. Its gene, aprE3-5, was cloned and expressed in a heterologous host (Jeong et al., 2007). In this study, the promoter of aprE3-5 was replaced with other stronger promoters (P_cry3A, P₁₀, P_SG1, P_srfA) of Bacillus spp. using PCR. The constructed chimeric genes were cloned into pHY300PLK vector, and then introduced into B. subtilis WB600. The P10 promoter conferred the highest fibrinolytic activity, i.e., 1.7-fold higher than that conferred by the original promoter. Overproduction of the 28 kDa protease was confirmed using SDS-PAGE and fibrin zymography. RT-qPCR analysis showed that aprE3-5 expression was 2.0-fold higher with the P10 promoter than with the original promoter. Change of the initiation codon from GTG to ATG further increased the fibrinolytic activity. The highest aprE3-5 expression was observed when two copies of the P₁₀ promoter were placed in tandem upstream of the ATG initiation codon. The construct with P10 promoter and ATG and the construct with two copies of P10 promoter in tandem and ATG exhibited 117% and 148% higher fibrinolytic activity, respectively, than that exhibited by the construct containing P10 promoter and GTG. These results confirmed that significant overproduction of a fibrinolytic enzyme can be achieved by suitable promoter modification, and this approach may have applications in the industrial production of AprE3-5 and related fibrinolytic enzymes.

• Preventive Nutrition and Food Science
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• 제17권1호
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• pp.78-82
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• 2012

Antithrombotic and fibrinolytic activity of natto was evaluated on platelet aggregation in vitro and in vivo. Natto showed inhibitory effects on platelet aggregation induced by adenosine 5’diphosphate (ADP) and collagen. Orally administered natto also showed fibrinolytic activity in hypercholesterolemia rats. Normal levels of natto, when administered for four weeks, shortened euglobulin clot lysis time (ECLT) and prolonged partial thromboplastin time (PATT) significantly compared to non-treated group. In addition, the natto treatment decreased total cholesterol in serum. These results showed that intake of normal levels of natto can elicit antithrombotic and fibrinolytic effects, suggesting its consumption may improve blood circulation.

• 한국균학회지
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• 제33권2호
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• pp.69-74
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• 2005

민자주방망이버섯으로부터 분리한 혈전용해효소의 비활성은 22.78 U/mg 이었으며, fibrin를 직접 용해하는 fibrinolytic enzyme 이었다. 15번째까지 N-terminal amino acid 서열 분석 결과, Tyr-Pro-Ser-Pro-Ser-His-Gln-Thr-Ala-Val-Asn-Ala-Ile-Ile-X로 지금까지 발표되지 않은 새로운 효소였다. 분자량은 34 KDa 이고 pH 7.0 부터 pH 9.5의 넓은 영역에서 높은 활성을 나타내는 alkaline protease 였으며, $55^{\circ}C$에서 가장 큰 활성을 보이는 이 효소는 serine protease 저해제인 phenylmethylsulfonyl fluoride를 첨가한 경우 효소의 활성이 전혀 나타나지 않는 것으로 미루어 높은 혈전용해 활성을 갖는 새로운 serine protease로 생각된다. 또한 $Hg^{2+}$의 금속이온을 첨가한 경우에도 효소의 활성은 완전히 사라졌다.

• 대한의생명과학회지
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• 제13권3호
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• pp.245-249
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• 2007

We investigated the fibrinolytic and ${\alpha}-glucosidase$ inhibitory activity of 55 wild mushroom methanol extracts. Among them, 14 mushrooms showed fibrinolytic activity. In particular, Amanita virgineoides showed the greatest enzyme activity (3.9 plasmin units/ml) by a fibrin plate assay. The fibrinolytic activities of Suillus pictus and Polypolellus varius were 3.8 plasmin units, and the activity of Gomphus fujisanensis was 2.8 plasmin units. Leccinum extremiorientale and Xerocomus nigromaculatus had the same activity with 2.3 plasmin units. In a ${\alpha}-glucosidase$ inhibitory activity test, Lactarius sp. showed the greatest inhibitory activity at 97.3%. The ${\alpha}-glucosidase$ inhibitory activities of Clitocybe odora, Xerocomus nigromaculatus, Melanoleuca melaleuca, Suillus pictus, and Gyroporus castaneus were 84.3%, 77.9%, 74.6%, 68.7%, and 65.4%, respectively. According to the results, because Suillus pictus and Xerocomus nigromaculatus have strong fibrinolytic and ${\alpha}-glucosidase$ inhibitory activities, the two mushrooms will be used as materials for the development of new biofunctional food.