• Title, Summary, Keyword: fibrinolytic activity

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Purification and Characterization of a New Fibrinolytic Enzyme of Bacillus licheniformis KJ-31, Isolated from Korean Traditional Jeot-gal

  • Hwang, Kyung-Ju;Choi, Kyoung-Hwa;Kim, Myo-Jeong;Park, Cheon-Seok;Cha, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1469-1476
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    • 2007
  • Jeot-gal is a traditional Korean fermented seafood and has long been used for seasoning. We isolated 188 strains from shrimp, anchovy, and yellow corvina Jeot-gal, and screened sixteen strains that showed strong fibrinolytic activities on a fibrin plate. Among those strains, the strain that had the largest halo zone was chosen and identified as Bacillus licheniformis by using 16S rDNA sequencing and an API CHB kit. The fibrinolytic activity of Bacillus licheniformis was characterized and designated as bpKJ-31. The active component of bpKJ-31 was identified as a 37 kDa protein, designated bacillopeptidase F, by internal peptide mapping and N-terminal sequencing. The optimum activity of bpKJ-31 was shown at pH 9 and $40^{\circ}C$, with a chromogenic substrate for plasmin. It had high degrading activity for the $B{\beta}$-chain and $A{\alpha}$-chain of fibrin(ogen), and also acted on thrombin, but not skim milk and casein. The amidolytic activity of bpKJ-31 was inhibited by 1 mM phenylmethanesulfonyl fluoride, but 1 mM EDTA did not affect the enzyme activity, indicating that bpKJ-31 is an alkaline serine protease, like a plasmin. The bpKJ-31 showed approximately 14.3% higher fibrinolytic activity than the plasmin. These features of bpKJ-31 make it attractive as a health-promoting biomaterial.

Purification and Biochemical Characterization of a Novel Fibrinolytic Enzyme from Streptomyces sp. P3

  • Cheng, Guangyan;He, Liying;Sun, Zhibin;Cui, Zhongli;Du, Yingxiang;Kong, Yi
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1449-1459
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    • 2015
  • A novel proteolytic enzyme with fibrinolytic activity, FSP3, was purified from the recently isolated Streptomyces sp. P3, which is a novel bacterial strain isolated from soil. FSP3 was purified to electrophoretic homogeneity by ammonium sulfate precipitation, anion exchange, and gel filtration. FSP3 is considered to be a single peptide chain with a molecular mass of 44 kDa. The maximum activity of the enzyme was observed at 50℃ and pH 6.5, and the enzyme was stable between pH 6 and 8 and below 40℃. In a fibrin plate assay, FSP3 showed more potent fibrinolytic activity than urokinase, which is a clinical thrombolytic agent acting as a plasminogen activitor. The activity was strongly inhibited by the serine protease inhibitor PMSF, indicating that it is a serine protease. Additionally, metal ions showed different effects on the activity. It was significantly suppressed by Mg2+ and Ca2+ and completely inhibited by Cu2+, but slightly enhanced by Fe2+. According to LC-MS/MS results, its partial amino acid sequences are significantly dissimilar from those of previously reported fibrinolytic enzymes. The sequence of a DNA fragment encoding FSP3 contained an open reading frame of 1287 base pairs encoding 428 amino acids. FSP3 is a bifunctional enzyme in nature. It hydrolyzes the fibrin directly and activates plasminogen, which may reduce the occurrence of side effects. These results suggest that FSP3 is a novel serine protease with potential applications in thrombolytic therapy.

Purification and Characterization of a Fibrinolytic Enzyme form Bacillus sp. KDO-13 Isolated from Soybean Paste

  • Lee, Si-Kyung;Bae, Dong-Ho;Kwon, Tae-Jong;Lee, Soo-Bok;Lee, Hyung-Hoan;Park, Jong-Hyun;Heo, Seok;Johnson, Michael-G.
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.845-852
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    • 2001
  • A microorganism producing fibrinolytic enzyme was isolated from Korean traditional soybean paste and identified as Bacillus sp. KDO-13. The fibrinolytic enzyme was purified to homogeneity by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-celluose, and gel chromatography on Sephadex G-100 of the culture supernatant of Bacillus sp. KDO-13. The molecular weight of the purified enzyme was estimated to be 44,000 by SDS-PAGE. The optimum pH and temperature for the enzyme activity were pH 8.0 and $50{\circ}C$, respectively. The enzyme activity was relatively stable at pH 7.0-9.0 and temperature below $50{\circ}C$. the activity of the enzyme was inhibited by $AI^{3+}$ and $Hg^{2+}$, but activated by $Co^{2+}$\;and\;Ni^{2+}. In addition, the enzyme activity was potently inhibited by EDTA and 0-phenanthroline. The purified enzyme could completely hydrolyze a fibrin substrate within 6 h in vitro, and had a low $K_m$ value for fibrin hydrolysis. It was concluded that the purified enzyme was a metalloprotease with relatively high specificity for fibrinolysis, and thus, could be applied as an effective thrombolytic agent.

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Characterization of the Functional Properties of Soy Milk Cake Fermented by Bacillus sp.

  • Oh, Soo-Myung;Kim, Chan-Shick;Lee, Sam-Pin
    • Food Science and Biotechnology
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    • v.15 no.5
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    • pp.704-709
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    • 2006
  • The mucilage production and tyrosine content in soy milk cake (SMC) fermented by Bacillus firmus NA-1, Bacillus subtilis GT-D, and B. subtilis KU-A was improved by fortification with 10% defatted soybean flour. The fibrinolytic activity and consistency of the SMC were drastically increased by solid-state fermentation for 1 day. However, the consistency of the fermented SMC gradually decreased during fermentation for 3 days. Furthermore, the tyrosine content of the freeze-dried powder of SMC fermented by three Bacillus sp. was 9 times higher than that of unfermented SMC. The soybean proteins, including the 7S and 11S subunits, were partially digested during alkaline fermentation, producing lower molecular-weight peptides. The fibrinolytic enzyme produced in SMC fermented by B. firmus NA-l and B. subtilis KU-A exhibited higher thermal stability than that of B. subtilis GT-D fermentation. The powder obtained from B. subtilis GT-D fermentation had an ${\alpha}$-amylase activity and lower consistency compared to those of B. firmus NA-1 and B. subtilis KU-A. In addition, this powder contained 6.3% moisture content, 27% crude protein content and 9 units of fibrinolytic activity and proteolytic activity.

Screening of Fibrinolytic Activities of Extracts from Wild Mushrooms Collected in Mt. Chilgap of Korea (칠갑산 야생버섯 추출물로부터 혈전용해효소의 활성 검색)

  • Kim, Jun-Ho;Yoo, Kwan-Hee;Seok, Soon-Ja;Kim, Yang-Sup
    • The Korean Journal of Mycology
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    • v.33 no.1
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    • pp.18-21
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    • 2005
  • Extracts of 67 wild mushrooms were tested for their fibrinolytic activities. The extract of Marasmius pulcherripes showed 112% higher increased activity as compared with a positive control, plasmin (0.75 U/ml). Helvella elastica and Psathyrella sp. showed 60% and 49% of activities to that of plasmin, respectively. Three mushrooms such as Lepista sordida, Fomitella fraxinea, and Leucoagaricus rubrotinctus showed 40% of plasmin activity. The other 60 mushroom extracts did not show any fibrinolytic activity.

A Correlation Between Fibrinolytic Activity and Microflora in Korean Fermented Soybean Products (된장의 혈전용해효소 활성과 미생물 분포와의 상관관계)

  • 김동호;송현파;김기연;김정옥;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.41-46
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    • 2004
  • A correlation between fibrinolytic activity and microflora in Korean traditional soybean fermented food was investigated. The fibrinolytic activities of traditional soybean pastes and commercially processed samples were 2.42$\pm$1.01 unit/g and 1.58$\pm$0.98 unit/g, respectively. The cell density of Bacillus in traditional soybean pastes was about 10$^{7}$ CFU/g and its commercially processed one was 10$^{6}$ CFU/g. Acid producing bacteria, fungi and yeast group were higher in commercially processed one. The correlations of fibrinolytic activity and microflora in traditional and commercial Doenjang were positively correlated in Bacillus ($R^2$≒ 0.69), negatively correlated in fungal group ($R^2$≒0.40), and there were no significant correlations in acid forming bacteria and yeast group ($R^2$<0.16). Fibrinolytic activities in Meju and Koji were 6.54$\pm$1.97 unit/g and 1.46$\pm$0.43 unit/g respectively, and were positively correlated with Bacillus. Yeast and acid forming bacteria were grown by 5∼6 decimal induction during fermentation period of Doenjang, but Bacillus, fungal cells and fibrinolytic activity were nearly stable. Results indicate that fibrinolytic activity of Doenjang depends on enzyme induction in Meju or Koji processing by Bacillus, Doenjang fermentation process.

Characterization and Fibrinolytic Activity of Acetobacter sp. FP1 Isolated from Fermented Pine Needle Extract

  • Park, Jae-Young;Yoon, Seo-Hyeon;Kim, Seong-Sim;Lee, Beom-Gi;Cheong, Hyeong-Sook
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.215-219
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    • 2012
  • The strain KCTC 11629BP, isolated from spontaneously fermented pine needle extract (FPE), showed fibrinolysis activity. The isolated strain was analyzed in physiological and biochemical experiments. Based on 16S rDNA sequencing and phylogenic tree analysis, the strain was identified to be a part of the genus Acetobacter, with Acetobacter senegalensis and Acetobacter tropicalis as the closest phylogenetic neighbors. Based on genotypic and phenotypic results, it was proposed that bacterial strain KCTC 11629BP represents a species of the genus Acetobacter. The strain was thusly named Acetobacter sp. FP1. In conclusion, Acetobacter sp. FP1 isolated from FPE possesses fibrinolytic activity.

Physiological Functionalities of Hot Water Extract of Codonopsis lanceolata and Some Medicinal Materials, and Their Mixtures (더덕과 일부 한약재 열수추출물의 혼합 비율에 따른 생리활성)

  • Oh, Hae-Sook;Kim, Jun-Ho
    • The Korean Journal of Community Living Science
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    • v.18 no.3
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    • pp.407-415
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    • 2007
  • The aim of this study was to investigate the physiological functionalities of Codonopsis lanceolata, Glycyrrhiza uralensis, Chaenomeles sinensis, Crataegus pinnafida, and their mixtures. We also determined their antioxidative, fibrinolytic, and ${\alpha}-glucosidase$ inhibitory activities. The antioxidative activities of Codonopsis lanceolata, Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida were 79%, 88.3%, 89.9%, and 89.3% respectively. Their fibrinolytic activities were 0.80plasmin unit/mi, 0.57 plasmin unit/mi, 0.52 plasmin unit/mi, and 0.53 plasmin unit/mi respectively. The ${\alpha}-glucosidase$ inhibitory activity of Codonopsis lanceolata was 25%. The 10-fold diluents of Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida showed ${\alpha}-glucosidase$ inhibitory activities of 93.6%, 65.3%, and 61.3% respectively. In antioxidative activity tests of the medicinal plants mixtures at various ratios, the mixtures of Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida with Codonopsis lanceolata showed antioxidative activities of approximately 90%. In fibrinolytic activity tests mixtures(1:1) of Codonopsis lanceolata with Chaenomeles sinensis and Crataegus pinnafida exhibited increases of 23% and 24% in activity respectively. In ${\alpha}-glucosidase$ inhibitory activity tests, a mixture (4:1) of Codonopsis lanceolata and 10-fold diluted Glycyrrhiza uralensis showed an inhibitory activity of 98%, a mixture (3:1) of Codonopsis lanceolata and 10-fold diluted Chaenomeles sinensis showed an inhibitory activity of 69.6%, and a mixture (1:1) of Codonopsis lanceolata and 10-fold diluted Crataegus pinnafida showed an inhibitory activity of 50.2%. In conclusion, the mixtures of Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida with Codonopsis lanceolata will be used as a material for the development of biofunctional foods.

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Development of Functional Doenjang for Antioxidative and Fibrinolytic Activity (항산화와 혈전용해 활성을 갖는 기능성 된장의 개발)

  • 류병호
    • Journal of Life Science
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    • v.13 no.5
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    • pp.559-568
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    • 2003
  • The aim of present study is to investigated on the development of the functional doenjang possessing antioxidative and fibrinolytic activity. A strain, BH-23 showing antioxidative and fibrinolytic activity was isolated from traditionally doenjang, and then identified as Bacillus subtilis based on moophological, physiological, and biochemical characteristics, and named as Bacillus subtilis BH-23. The optimal pH level and temperature conditions were 5.0 and $40^{\circ}C$, NaCl concentration was grown well at 15% Antioxidative and fibrinolytic activity of Bacillus subtilis BH-23 was accoidance with growth conditions of the original state. By comparising with traditionally commercialized doenjang, it is prepared with Bacillus subtilis BH-23. The final results evaluated with positive elements rised above the surface such as color, taste and flavor without ammonia. Therefore, it is safe to assume, doenjang has strong market value in the food industry.

Fibrinolytic Activity and Chemical Properties of Cordycepin-Enriched Cordyceps militaris JLM 0636 (Cordycepin 고함유 동충하초 수용성 추출물의 이화학적 특성과 혈전 용해활성)

  • Ahn, Hee-Young;Lee, Jae-Hong;Kang, Min-Jeong;Cha, Jae-Young;Cho, Young-Su
    • Journal of Life Science
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    • v.22 no.2
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    • pp.226-231
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    • 2012
  • The effect of cordycepin-enriched Cordyceps militaris JLM 0636 ($CM{\alpha}$) and Cordyceps militaris (CM) on fibrinolytic activity was investigated. The bioactive compounds and nutritional materials such as polyphenolic compounds, flavonoids, glutathione, minerals, and fatty acids were also measured. Concentrations of polyphenol compounds, flavonoids, and glutathione were higher in $CM{\alpha}$ than that in CM. The major minerals of both materials were K, Ca, Mg, and Na. The major fatty acids of both materials were linolenic acid, linoleic acid, oleic acid, and palmitic acid. Fibrinolytic activity was higher in $CM{\alpha}$ than that in CM. These results may provide the basic data to understand the fibrinolytic activity and bioactive compounds of $CM{\alpha}$.