Purification and characterization of a new fibrinolytic enzyme of Bacillus licheniformis KJ-31, isolated from Korean traditional Jeot-gal.
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 degrees C, with a chromogenic substrate for plasmin. It had high degrading activity for the Bbeta-chain and Aalpha-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.
Hwang
Kyung-Juw
KJ
Department of Microbiology, College of Natural Sciences, Pusan National University, Busan 609-735, Korea.
Choi
Kyoung-Hwa
KH
Kim
Myo-Jeong
MJ
Park
Cheon-Seok
CS
Cha
Jaeho
J
D001407:Bacillus / Q000201:enzymology*; D005337:Fibrin / Q000378:metabolism; D005343:Fibrinolytic Agents / Q000737:chemistry / Q000302:isolation & purification*; D005516:Food Microbiology; D007723:Korea; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification*; D013379:Substrate Specificity; D013917:Thrombin / Q000378:metabolism
D005343:Fibrinolytic Agents; D005337:Fibrin; D012697:Serine Endopeptidases; C038477:bacillopeptidase F; D013917:Thrombin