Browse > Article
http://dx.doi.org/10.3746/jfn.2006.11.2.127

Purification and Characteristics of Fibrinolytic Enzyme from Chongkukjang  

Yang, Jeong-Lye (School of Food and Life Science, Biohealth Product Research Center and Food Science Institute, Inje University)
Kim, Hee-Sook (Department of Food Science and Technology, Kyungsung University)
Hong, Jeong-Hwa (School of Food and Life Science, Biohealth Product Research Center and Food Science Institute, Inje University)
Song, Young-Sun (School of Food and Life Science, Biohealth Product Research Center and Food Science Institute, Inje University)
Publication Information
Preventive Nutrition and Food Science / v.11, no.2, 2006 , pp. 127-132 More about this Journal
Abstract
Bacillus sp. strain K-l, which produces a strong fibrinolytic enzyme, was isolated from chongkukjang, a traditional Korean fermented soybean paste. The fibrinolytic enzyme was purified from chongkukjang base by using ammonium sulfate fractionation and chromatographic techniques. Purified enzyme, CK K-1 was demonstrated to be homogeneous by SDS-PAGE and isoelectric focusing electrophoresis, and has molecular mass of a 12.4 kDa and a pI of 8.0. The optimal reaction pH value and temperature were 8.0 and $40^{\circ}C$, respectively. Phenyl-methyl-sulfonyl-fluoride (PMSF; serine protease inhibitor), ethylene-diamine-tetra-acetic acid (EDTA; metallo protease inhibitor), copper ion, ferric ion and lead ion inhibited the enzyme activity. These results indicated that the fibrinolytic enzyme is a metallo-serine protease and different from nattokinase and chongkukjangkinase.
Keywords
fibrinolytic enzyme; chongkukjang; soybean; chromatography; zymography;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Voet D, Voet JG. 1990. Biochemistry. John Wiley & Sons, New York. p 1087-1095
2 Wun TC, Schleuning WD, Reich E. 1982. Isolation and characterization of urokinase from human plasma. J Biol Chem 257: 3276-3283
3 Medved LV, Solovjov DA, Ingham KC. 1996. Domain structure, stability and interaction in streptokinase. Eur J Biochem 239: 333-339   DOI   ScienceOn
4 Pennica D, Holmes WE, Kohr WJ, Harkins RN, Vehar GA, Ward CA, Bennn WF, Yelverton E, Seeburg PH, Heyneker HL, Goeddel DV, Cohen D. 1982. Clonning and expression of human tissue-type plasminogen activator cDNA in E. coli. Nature 301: 214-221   DOI   ScienceOn
5 Sumi H, Hamada H, Nakanishi K, Hiratani H. 1990. Enhancement of the fibrinolytic activity in plasma by oral administration of nattokinase. Acta Haematol 84: 139-143   DOI
6 Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254   DOI   ScienceOn
7 Astrup T, Mullertz S. 1952. The fibrin plate method for estimating fibrinolytic activity. Arch Biochem Biophys 40: 346-351   DOI   ScienceOn
8 Laemmli UK. 1970. Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227: 680-685   DOI   ScienceOn
9 Kim SH, Choi NS. 1999. Electrophoretic analysis of protease inhibitors in fibrin zymography. Anal Biochem 270: 179-181   DOI   ScienceOn
10 Hong JH, Youn HK, Kang MC, Lee HJ, Hur SH. 2006. Properties of Bacillus spp. isolated from fermenting chongkukjang. J Korean Soc Ind Food Technol 4: 67-72
11 Holt JG, Sharpe ME, Mair NS, Sneath PH. 1989. Bergey's Manual of Systematic Bacteriology. Williams and Wilkins press, Philadelphia. Vol 2, p 1105
12 Nakajima N, Tayu N, Sumi H. 1993. Potentfibrinolytic enzyme from the lysate of Katsuwohus pelamis digestive tract (Shiokara): purification and characterization. Biosci Biotech Biochem 57: 1604-1605   DOI   ScienceOn
13 Kim SH, Choi NS. 2000. Purification and characterization of subtilisin DJ-4 secreted by Bacillus sp. strain DJ-4 screened from Doen-Jang. Biosci Biotechnol Biochem 64: 1722-1725   DOI   ScienceOn
14 Noh KA, Kim DH, Choi NS, Kim SH. 1999. Isolation of fibrinolytic enzyme producing strains from kimchi. Korean J Food Sci Technol 31: 219-223   과학기술학회마을
15 Kim JH, Kim YS. 1999. A fibrinolytic metalloprotease from the fruiting bodies of an edible mushroom, Armillariella mellea. Biosci Biotechnol Biochem 63: 2130-2136   DOI   ScienceOn
16 Kim HK, Kim GT, Kim DK, Choi WA, Park SH, Jeong YK, Kong IS. 1997. Purification and characterization of a novel fibrinolytic enzyme from Bacillus sp. KA38 orig-inated from fermented fish. J Ferment Bioeng 84: 307-312   DOI   ScienceOn
17 Whitaker JR. 1994. Enzyme inhibitor. In Principles of Enzymology for the Food Science. 2nd ed. Whitaker JR, ed. Dekker, New York. p 241-270
18 Sumi H, Hamada H, Tsushima H, Mihara H, Muraki H. 1987. A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese natto: a typical and popular soybean food in the Japanese diet. Experientia 43: 1110-1111   DOI   ScienceOn
19 Kim W, Choi K, Kim Y, Park H, Choi J, Lee Y. 1996. Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. strain CK 11-4 screened from Chungkuk-jang. Appl Environ Microbiol 62: 2482-2488