• Title, Summary, Keyword: fibrinolytic activity

Search Result 312, Processing Time 0.058 seconds

Partial Purification and Characterization of Fibrinolytic Substance from Wooltalikong (Phaseolus ssp.)

  • Oh Hae-Sook;Kim Jun-Ho
    • Biomedical Science Letters
    • /
    • v.10 no.4
    • /
    • pp.415-420
    • /
    • 2004
  • Fibrinolytic substance was purified from the Wooltalikong (Phaseolus ssp.), using DEAE-cellulose chromatography, Sephadex G-150 gel-filtration, and FPLC gel-filtration. The substance has a molecular weight of 5262.70 Da as measured by MALD-TOF mass spectrometry. It has a pH optimum at pH 6.0. The fibrinolytic activity of purified substance was inhibited by EDTA and 1,10-phenanthroline and slightly decreased by PMSF and pepstatin A. It shows the maximum fibrinolytic activity at 40℃ and the substance was stable up to 50℃. The activity of the substance was increased by Zn/sup 2+/ and was totally inhibited by Hg/sup 2+/. This study revealed that Wooltalikong could be a good source of fibrinolytic products due to its small molecular size and heat-resistant ability.

  • PDF

Characterization of a Fibrinolytic Enzyme Secreted by Bacillus velezensis BS2 Isolated from Sea Squirt Jeotgal

  • Yao, Zhuang;Kim, Jeong A;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
    • /
    • v.29 no.3
    • /
    • pp.347-356
    • /
    • 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.

Analysis of Biological Activities of Medicinal Mushrooms (약용버섯의 생리활성 분석)

  • Hur, Hyun
    • Korean Journal of Pharmacognosy
    • /
    • v.39 no.3
    • /
    • pp.265-269
    • /
    • 2008
  • This study was carried out to observe the antioxidative activity, fibrinolytic activity, nitrite savenging ability and adenosine content of three medicinal mushrooms. Among water and ethanol extract of each mushroom, water extract of Inonotus obliquus (Czech) showed the highest antioxidative activity $(57.1{\mu}g/ml)$, nitrite scavenging ability (52.04%), fibrinolytic activity (86.8%) and adenosin content $(94.3{\mu}g/g)$, whereas nitrite scavenging ability of ethanol extract of Phellinus linteus showed higher than that of water extract. Apart from the above statements, water was effective than ethanol as extracting solvent in general. These results suggest that Inonotus obliquus (Czech) showed higher activities such as antioxidative activity, nitrite scavenging ability, fibrinolytic activity and adenosin content and water was effective as solvent except for nitrite scavenging ability.

Effects of Insect Crude Drugs on Blood Coagulation and Fibrinolysis System

  • Ahn, Mi-Young;Hahn, Bum-Soo;Ryu, Kang-Sun;Cho, Sung-Ig
    • Natural Product Sciences
    • /
    • v.8 no.2
    • /
    • pp.66-70
    • /
    • 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.

Identification and Characterization of Fibrinolytic Compound from Cornus officinalis S. et Z (산수유(Cornus officinalis)로부터 혈전용해물질의 확인 및 특성 연구)

  • Kim, Jun-Ho
    • Korean Journal of Plant Resources
    • /
    • v.33 no.4
    • /
    • pp.237-244
    • /
    • 2020
  • The objective of this study was to identify and characterize fibrinolytic compound from Cornus officinalis. Cornus officinalis. Hot water extract was fractionated into hexane, chloroform, ethyl acetate, butanol, and water fractions. Assays for fibrinolytic activity indicated that only the ethyl acetate fraction had significant efficacy at 1.36 plasmin units/mL. Isolation of fibrinolytic compound was carried out on Amberlite IRA-400, Sephadex LH-20 and active charcoal column chromatography. HPLC analysis of the purified fibrinolytic compound showed retention time (RT) same as authentic malic acid. LC / MS / MS in negative mode showed the same peak at m/z 133, confirming that the purified compound was malic acid with a molecular weight 134 Da. The compound showed fibrinolytic activity of 0.69 plasmin units/mL, 14.62% of thrombin inhibitory activity, 6.42% of antioxidative activity, and 17.28% of α-glucosidase inhibitory activity. The purified compound hydrolyzed γ subunits of human fibrinogen. In conclusion, malic acid isolated from Cornus officinalis might have potential to be developed as ingredient for biofunctional foods to prevent cardiovascular diseases.

Screening and Characterization of Microorganisms with Fibrinolytic Activity from Fermented Foods

  • Yoon, Seon-Joo;Yu, Myeong-Ae;Sim, Gwan-Sub;Kwon, Seung-Taek;Hwang, Jae-Kwan;Shin, Jung-Kue;Yeo, In-Hyun;Pyun, Yu-Rang
    • Journal of Microbiology and Biotechnology
    • /
    • v.12 no.4
    • /
    • pp.649-656
    • /
    • 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.

Variation of fibrinolytic enzyme activity produced Bacillus subtilis by gene cloning (유전자 cloning에 의한 Bacillus subtilis의 fibrinolytic enzyme 활성 변화)

  • 이홍석;유천권;이철수;강상모
    • Microbiology and Biotechnology Letters
    • /
    • v.28 no.1
    • /
    • pp.14-20
    • /
    • 2000
  • The transformation of Bacillus subtilis K-54 and J-10 was carried out with constructed vectors containing structure and enhancer genes of aprN and prtR, to increase their fibrinolytic enzyme activity. Bands for the aprN and prtR genes were identified from B. subtilis J-10 by PCR that was carried out with the constructed primers for the genes. In addition, the gene fragments contained promoter site based on the results of analysing their nucleotide sequence. The two gene fragments, aprN and prtR, obtained by the PCR, were, then, inserted to vector such as T-vector and E.coli/Bacillus shuttle vector. The constructed vector were designated as pAPR2 (aprN), pENC2 (prtR) and pFLA1 (aprN and prtR), respectively. The constructed vector was used for transformation of the strains of B.subtilis J-10 and B. subtilis K-54 and the fribrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and the fibrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and pFLA1, resulted in the increase of fibrinolyitic enzyme activity in B. subtilis J-10 by 27.3% and 16%, respectively. However, the introduction of pENC2 to B. subtilis J-10 did not seem to induce increase of the enzyme activity. The strain of B.subtilis K-54 transformed with pENC2 showed an increased fibrinolytic activity by 5 folds compared with that of the original strain of B. subtilis K-54.

  • PDF

Pheophytin Content and Fibrinolytic Activity of Silkworm Feces in the Different Larval Stages of Silkworms

  • Ahn, Mi-Young;Ryu, Kang-Sun;Kim, Iksoo;Kim, Jin-Won;Lee, Heui-Sam;Lee, Yong-Ki;Kim, Eun-Sun
    • International Journal of Industrial Entomology
    • /
    • v.5 no.2
    • /
    • pp.195-199
    • /
    • 2002
  • In order to find potential anticancer agents, we extracted pheophytin in the silkworm feces from various larval stages by water, chloroform and methanol extraction. The cytotoxicity of the pheophytin extracts of various silkworm feces was measured in the CT-26 cells originated from murine metastatic colon cancer, by dye uptake assay. The cytotoxicity of those pheophytins in 2nd, 3rd and 4th instars was better than remaining larval stages. The in vitro anticoagulant and fibyinolytic activities of ethanol extract from varietal mulberry leaves, mulberry branches and silkworm feces and pheophytin extracts from silkworm feces obtained at various larval stages were evaluated in order to find effective therapeutic drugs for the treatment of myocardial and cerebral thrombosis. The fibrinolytic activity was tested using the activated partial thromboplastin time (APTT) and thrombin time (TT) was measured for blood clotting activity. With regards to the fibrinolytic system, ethanol extracts of silkworm feces were better than varietal mulberry leaves and mulberry branches. The pheophytin extracts from 7th days of 5th instar contained the highest percentage of pheophytin and good fibrinolytic activity.

Characterization of Antithrombotic Activity of Lumbrokinase immobilized Polyurethane Heart Valves in Total Artificial Heart Experiment

  • Park, Y.D.;Jeong, J.S.;Kim, H.J.;Kim, J.;Min, B.G.
    • Proceedings of the KOSOMBE Conference
    • /
    • v.1997 no.05
    • /
    • pp.51-54
    • /
    • 1997
  • Lumbrokinase, potent fibrinolytic enzyme purified from earthworm, was immobilized onto the total artificial heart valves using photoreaction. This valve were implanted into the lamb for three days. After experiments, thrombus was observed in the untreated valves whereas no thrombus was observed in the lumbrokinase immobilized valves. The fibrinolytic activity and proteolytic activity of the implanted valve was examined. The fibrinolytic activity of the valve was remained after the implantation. The lumbrokinase could be a suitable fibrinolytic agents in the vascular contacting devices to reduce the thrombus.

  • PDF

Fibrinolytic Enzyme Production by Bacillus subtilis KH-4 Isolated from Deonjang

  • Kim, J.M.;Suh, H.J.;Ahn, S.W.;Kim, M.S.;Oh, S.H.
    • Preventive Nutrition and Food Science
    • /
    • v.7 no.4
    • /
    • pp.417-420
    • /
    • 2002
  • A strong fibrin-specific fibrinolytic enzyme was produced from Bacillus subtilis KH-4 isolated from Deonjang, a Korean fermented soybean paste similar to Japanese miso. The addition of glucose as a carbon source resulted in the highest levels of caseinolytic and fibrinolytic activities. Likewise, the addition of yeast extract as the nitrogen source resulted in the highest caseinolytic and fibrinolytic activities (3473.2 unit and 47.4 munit, respectively), It was observed that out of all metal ion sources only calcium (chloride) enhanced caseinolytic and fibrinolytic activities, with increases of 4949.3 unit and 58.2 unit/mg, respectively. The optimal temperature for the production of the enzyme was found to be 4$0^{\circ}C$ in the optimal medium (glucose 20 g, yeast extract 5 g, CaCl$_2$l g, and NaCl 2 g). The maximum fibrinolytic activity was observed at the late stationary phase. B. subtilis KH-4 produced a fibrinolytic enzyme at 4$0^{\circ}C$, after 30 h growth, which increased up to 54 h and then remained constant. These results suggest that Deonjang has potential as a source of physiologically active anti-thromotic enzymes.