Purification and Characterization of the FibrinolyticEnzyme Produced by Bacillus subtilis KCK-7 from Chungkookjang

• Published in: Journal of microbiology and biotechnology pp. 829 - 835
• Main Authors: PAIK, HYUN-DONG, SI-KYUNG LEE, SEOK HEO, SOO-YOUNG KIM, HYUNG-HOAN LEE, TAE-JONG KWON
• Format: Journal Article
• Language: Korean
• Published: 한국미생물·생명공학회 01.08.2004
• Subjects: 생물학
• ISSN: 1017-7825

A fibrinolytic enzyme has been found in severalbacteria isolated from fermented food. This study was carriedout to investigate the purification and characteristics of thefibrinolytic enzyme produced by Bacillus subtilis KCK-7originated from Chungkookjang. The fibrinolytic enzymewas purified to homogeneity from the culture supernatantusing ammonium sulfate fractionation and chromatographieson DEAE-cellulose and on Sephadex G-100. The finalspecific activity of the purified enzyme increased 11.0-fold,and the protein amount in the purified enzyme was about 16%of that in the culture supernatant. The molecular weight of thepurified enzyme was estimated to be about 45,000 by SDSPAGE.The optimum pH and temperature for the enzymeactivity were pH 7.0 and 60oC, respectively. The enzymeactivity was relatively stable up to 60oC over the pH range of7.0- 10.0. The fibrinolytic enzyme activity increased by Ca2+and Cu2+, whereas it was inhibited by Hg2+ and Ba2+. Inaddition, it was severely inhibited by PMSF and DFT. Itis suggested that the purified enzyme was a serine proteasefor the fibrinolysis. The purified enzyme could completelyhydrolyze fibrin in vitro within 8 h. Hence, it is suggested thatthe purified enzyme can be put into practice as an effectivethrombolytic agent. KCI Citation Count: 41