• Title/Summary/Keyword: Fibrin plate

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The Fibrinolytic Activity of Kimchi and its Ingredients in vivo and in vitro (In vivo와 in vitro에서 김치 및 김치재료의 피브린 분해활성)

  • 송영옥;김미정;송영선
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.4
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    • pp.633-638
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    • 1998
  • Fibrionolytic activity(FA) of kimchi in rat and FAs of water and methanol extracts of kimchi ingredient were determined using fibrin plate method to see where the active principles for FA are present in kimchi. Nine Sprague-Dawley per each group were fed diet containing 3, 5 or 10% of freeze dried kimchi for 6 weeks. The FA of plasma obtained from 10% kimchi group showed stronger FA than that of control. FAs of 3 and 5% kimchi group were not noticeable. FA from methanol extract of kimchi was approximately 6 times higher than that from water extract assuming that the active material may be present in a fat soluble fraction. From water extract of kimchi ingredients, dropwort, green onion, radish and baechu showed FA in descending order. And from methanol extracts, red pepper powder, radish, green onion and baechu showed FA in decending order. The Fa of methanol extract of red pepper powder was the most noticeable among samples.

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Studies on the Biological Activity of Astragalus membranaceus Extracts

  • Kim, Jun-Ho
    • Biomedical Science Letters
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    • v.18 no.1
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    • pp.35-41
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    • 2012
  • Physiological activities of hot water extract and solvent fractions isolated from Astragalus membranaceus were examined and the antioxidative, fibrinolytic, thrombin inhibitory and a-glucosidase inhibitory activity were measured. The hot water extract of Astragalus membranaceus was fractionated into hexane, chloroform, ethyl acetate, butanol and water fractions, and each of these fractions was individually assayed. The antioxidative activities of ethyl acetate and chloroform fractions were 89.96% and 87.36%, respectively. Using the fibrin plate method, only the hot water extract showed a plasmin activity of 0.41 units/ml. The thrombin inhibitory activity of the ethyl acetate fraction was the highest with a value of 82.73%. The hot water extract displayed a-glucosidase inhibitory activity of 64.91%. In conclusion, the hot water extract and the ethyl acetate fraction can be used as materials for the development of biofunctional foods to prevent cardiovascular diseases.

A Novel Anticoagulant Protein from Scapharca broughtonii

  • Jung, Won-Kyo;Je, Jae-Young;Kim, Hee-Ju;Kim, Se-Kwon
    • BMB Reports
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    • v.35 no.2
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    • pp.199-205
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    • 2002
  • An anticoagulant protein was purified from the edible portion of a blood ark shell, Scapharca broughtonii, by ammonium sulfate precipitation and column chromatography on DEAE-Sephadex A-50, Sephadex G-75, DEAE-Sephacel, and Biogel P-l00. In vitro assays with human plasma, the anticoagulant from 'S. broughtonii, prolonged the activated partial thromboplastin time (APTT) and inhibited the factor LX in the intrinsic pathway of the blood coagulation cascade. But, the fibrin plate assay did not show that the anticoagulant is a fibrinolytic protease. The molecular mass of the purified S. broughtonii anticoagulant was measured to be about 26.0kDa by gel filtration on a Sephadex G-75 column and SDS-PAGE under denaturing conditions. The optimum activity in the APTT assay was exhibited at pH 7.0-7.5 and $40-45^{\circ}C$ in the presence of $Ca^{2+}$.

Anti-thrombosis Activity of Drone Apis mellifera Pupae Extracts

  • Choi, Hong Min;Moon, Hyo Jung;Kim, Se Gun;Jang, Hye Ri;Woo, Soon Ok;Bang, Kyeong Won;Han, Sang Mi
    • Journal of Apiculture
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    • v.33 no.4
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    • pp.303-306
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    • 2018
  • Drones of honeybee (Apis mellifera) have been regarded as a useful value only when mating with queen bee. However, the drone pupae have been reported to be nutritionally valuable, and a potential beekeeping product. In this study, drone pupae extracted with 5% acetic acid were used to measure anti-thrombosis related fibrinolytic activity using Strup and Mullertz fibrin plate method. As a result, the drone pupae extract showed higher effect of fibrinolytic activity(clear zone diameter 20.83mm) compared to the human plasmin (clear zone diameter 12.93mm) used as a positive control. It was suggested that the extract of drone pupae can be developed as a functional material helping prevention or treatment of various vascular diseases.

Fibrinolysis of Fermented Kimchi (김치의 혈전용해작용)

  • Jeong, Yong-Kee;Yang, Woong-Suk;Kang, Jeong-Ok;Kong, In-Soo;Kim, Jeong-Ok
    • Journal of Life Science
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    • v.5 no.4
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    • pp.203-210
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    • 1995
  • Fibrinolytic activity of fermented kimchi and ingredients used for the preparation were assayed by measuring the lysis area on plasminogen-rich fibrin plate. Fermented kimchi and picked fish sauces from changlan, prwan, and anchovy showed the activity, and the activity of pickled fish sauces were high in the order of pickled changlan, picked anchovy(pickled mulchi), and pickled prawn. However, the activity of kimchi may not be attributed to pickled fish sauce because kimchi containing fish sauce did not possess activities were determined from the samples heated for 30min at 100$\circ$C. There was no changes in activities before and after heat treatment indicating the agents may be components other than protein. Since major changes occur during kimchi fermentation were increased sour taste due to production of organic acids such as lactic, citric, oxalic, and succinic acids, the authentic organic acids present were examined for fibrinolytic activities. The results indicates that the major component posses the activity is lactic acid.

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Cloning and High Expression of Nattokinase Gene from Bacillus subtilis BB-1 (Bacillus subtilis BB-1으로부터 나토키나아제 유전자 크로닝 및 대량발현)

  • Lee Young-Hoon;Lee Sung-Ho;Park Ki-Hoon;Choi Young-Ju;Jeong Yong-Kee;Gal Sang-Wan
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.274-281
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    • 2006
  • A fibrinolytic enzyme gene was isolated from Bacillus subtilis BB-1 by PCR method. Primers for PCR cloning were designed according to pre-identified gene for fibrinolytic enzymes from B. subtilis. The primer sequences were 5'-CGG ATC CGT GAG AGG CAA AAA GGT G-3' and 5'-TGA ATT CTT AAT GTG CTG CTG CTT GTC C-3' as concensus sequences of the fibrinolytic genes of Bacillus species. The PCR product was 1,145 bp and the sequence homology was 99% with nattokinase gene isolated from Japanese natto. The cloned fibrinolytic gene was reconstructed in Bacillus-E. coli shuttle vector, pEB for bulk-production. The fibrinolytic enzyme was purified by FPLC from the cloned B. subtilis 168. The optimum pH and temperature of the enzyme were 7.0 and $35^{\circ}C$, respectively. The fibrinolytic enzyme did not show any activity toward to skim milk, gelatin, casein and blood agar plate. The enzyme specific polyclonal antibody was prepared in rabbit for further assays such as detection of the gene expression in plant cells. This means that the enzyme may be used for health-care such as thrombosis without any hamful effects in the blood vessel.

Antioxidant Activity, Fibrinolysis and Angiotensin I Converting Enzyme Inhibitory Activity of Pine Mushroom Juice (Tricholoma matsutake Sing) (송이즙의 항산화 활성, 혈전용해활성 및 Angiotensin I Converting Enzyme의 저해활성 검색)

  • Kim, Young-Eon;Kwon, Eun-Kyung;Han, Dae-Seok;Kim, In-Ho;Ku, Kyung-Hyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.5
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    • pp.535-541
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    • 2008
  • Pine mushroom (Tricholoma matsutake Sing.) is an expensive and highly prized delicacy in Korean and Japanese cuisines with its unique flavor and functional properties. The biological activities of pine mushroom juice (soluble solid contents $4.3^{\circ}$Brix) were evaluated using different tests; DPPH radical scavenging assay for its antioxidant activity, fibrin plate method for fibrinolysis and angiotensin I converting enzyme (ACE) inhibitory activity for anti-hypertensive effect. Free radical scavenging activity of the pine mushroom juice was $48.3{\pm}2.2%$ at the concentration of 1.0 mg/mL. The fibrinolytic activity of pine mushroom was about 2 times greater than that of plasmin used as positive control and the activity increased dose-dependently. The pine mushroom juice inhibited ACE activities dose-dependently and $IC_{50}$ value of ACE activity was $1.03^{\circ}$Brix. These results suggest that pine mushroom is a healthy delicacy.

Characteristics of Chungkookjang Produced by Bacillus subtillus MC31 (B. subtilis MC 31를 이용한 청국장의 품질특성)

  • Mann, So-Yon;Kim, Eun-Ah;Lee, Ga-Young;Kim, Ro-Ui;Hwang, Dae-Youn;Son, Hong-Joo;Lee, Byong-Won;Lee, Chung-Yeol;Kim, Dong-Seob
    • Journal of Life Science
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    • v.23 no.4
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    • pp.560-568
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    • 2013
  • Chungkookjang was fermented by B. subtilis MC31, a ${\gamma}$-amino butyric acid (GABA) producing microorganism. The characteristics of Chungkookjang were investigated while fermenting. Twenty four amino acids were detected in Chungkookjang, leucine was the highest of them all. Total cell populations of B. subtilis MC31 phase were between log $9.52{\pm}0.5$ ~ log $9.049{\pm}0.5$ CFU/g at stationary phase. Contents of moisture, crude ash, crude protein, crude lipid and crude fiber are $61.07{\pm}0.01%$, $1.52{\pm}0.01%$, $17.66{\pm}0.04%$, $8.96{\pm}0.03%$ and 2.61%, respectively. Contents of ammonia type nitrogen, amino type nitrogen and reducing sugar were increased during fermentation at $40^{\circ}C$ for 72 hr, however those of titratable acidity and total sugar were decreased. pH was slowly alkalized during fermentation. Viscous substance and protease contents in Chungkookjang were $4.7{\pm}0.05%$ and $0.519{\pm}7.36$ g/l, apiece. When the fibrin plate and Robbin method for fibrinolytic activity were applied, B. subtilis MC31 showed high activity. These results suggested that B. subtilis MC31 is suitable to be used as a starter to enhance the quality of Chungkookjang.

Isolation of Bacteria with Protease Activity from Cheonggukjang and Purification of Fibrinolytic Enzyme (청국장으로부터 혈전용해 활성이 우수한 균주 분리 및 혈전용해효소정제)

  • Choi, Yeon Hee;Lee, Jun Seung;Bae, So Young;Yang, Keun Jae;Yeom, Kyu Won;Jo, Dong Hyeok;Kang, Ock Hwa;Baik, Hyung Suk
    • Journal of Life Science
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    • v.23 no.2
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    • pp.259-266
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    • 2013
  • To isolate the fibrinolytic enzyme, 268 strains from 21 samples were morphologically isolated from Cheonggukjang collected from Korea and Japan. Among the 268 strains, protease-producing bacteria were isolated in nutrient agar medium including 1% skimmed milk. As a result of this, 22 strains were isolated. Apiweb site was used to identify these strains based on their biochemical properties. In addition, 16S rRNA sequencing was performed to identify the strain. Most of the identified strains were Bacillus subtilis and B. amyloliquefaciens. Fibrinolytic enzyme activity was measured with the fibrin plate method. Five strains were finally selected: A2-14, A2-20, C1-05, C1-09, and F2-01. Of those five strains, the A2-20 strain, which is close to B. amyloliquefaciens, showed the strongest fibrinolytic activity. The fibrinolytic enzyme produced by the A2-20 strain was partially purified from culture supernatant by gel filtration and ion exchange chromatography. The optimal pH and temperature values of the partially purified enzyme were 7.0 and $35^{\circ}C$, respectively. Purified protein analysis was carried out with SDS-PAGE and zymography. A genetic analysis was also conducted by PCR based on the consensus sequence of fibrinolytic enzyme. Corresponding genes with a partial sequence of the A2-20 strain were identified.

Pharmacetical Characteristics of Solvent Fractions Isolated from Glycyrrhiza uralensis

  • Kim, Jun-Ho;Oh, Hae-Sook
    • Biomedical Science Letters
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    • v.16 no.3
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    • pp.161-167
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    • 2010
  • In this study, the biological activities of Glycyrrhiza uralensis were investigated, including antioxidative, fibrinolytic, thrombin inhibitory, and a-glucosidase inhibitory activity. The hot water extract of Glycyrrhiza uralensis was fractionated into hexane, $CHCl_3$, ethyl acetate, butanol, and water fractions, and each of these fractions were assayed individually. The water fraction showed the highest extraction yield of 19.45% (w/w). Using the DPPH method, the free radical scavenging activity was to be the strongest in the $CHCl_3$ fraction at 89.3%. Using the fibrin plate method, only the butanol fraction showed a substantial plasmin activity of 0.62 units/ml. In thrombin inhibitory activity tests, a 100-fold dilution of the ethyl acetate fraction showed the strongest activity of 46.9%. In the a-glucosidase inhibitory activity assay, a 100-fold dilution of the $CHCl_3$ fraction showed the strongest activity of 80.6%. In conclusion, the combined results of this study demonstrate that the extracts of Glycyrrhiza uralensis can be used as a material for the development of biofunctional foods for diabetics.