title: Characterization of a Novel Fibrinolytic Enzyme, BsfA, from Bacillus subtilis ZA400 in Kimchi Reveals Its Pertinence to Thrombosis Treatment.
abstract: Recently, the cardiovascular disease has been widely problematic in humans probably due to fibrin formation via the unbalanced Western style diet. Although direct (human plasmin) and indirect methods (plasminogen activators) have been available, bacterial enzyme methods have been studied because of their cheap and mass production. To detect a novel bacterial fibrinolytic enzyme, 111 bacterial strains with fibrinolytic activity were selected from kimchi. Among them, 14 strains were selected because of their stronger activity than 0.02 U of plasmin. Their 16S rRNA sequence analysis revealed that they belong to Bacillus, Leuconostoc, Propionibacterium, Weissella, Staphylococcus, and Bifidobacterium. The strain B. subtilis ZA400, with the highest fibrinolytic activity, was selected and the gene encoding fibrinolytic enzyme (bsfA) was cloned and expressed in the E. coli overexpression system. The purified enzyme was analyzed with SDS-PAGE, western blot, and MALDI-TOF analyses, showing to be 28.4 kDa. Subsequently, the BsfA was characterized to be stable under various stress conditions such as temperature (4-40°C), metal ions (Mn(2+), Ca(2+), K(2+), and Mg(2+)), and inhibitors (EDTA and SDS), suggesting that BsfA could be a good candidate for development of a novel fibrinolytic enzyme for thrombosis treatment and may even be useful as a new bacterial starter for manufacturing functional fermented foods.
authors.affiliation: Department of Food Science and Biotechnology, Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea.
authors.forename: Min-Ju
authors.initials: MJ
authors.lastname: Ahn
authors.affiliation: Department of Food Science and Biotechnology, Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea.
authors.forename: Hye-Jin
authors.initials: HJ
authors.lastname: Ku
authors.affiliation: Department of Food Science and Biotechnology, Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea.
authors.forename: Se-Hui
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Department of Food Science and Biotechnology, Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea.
authors.forename: Ju-Hoon
authors.initials: JH
authors.lastname: Lee
mesh_terms: D001412:Bacillus subtilis / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005341:Fibrinolysin / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005516:Food Microbiology; D015870:Gene Expression; D006801:Humans; D008970:Molecular Weight; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013927:Thrombosis / Q000188:drug therapy
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D011994:Recombinant Proteins; D005341:Fibrinolysin
keywords: Bacillus subtilis; Fermented food; Fibrinolytic activity; Gene expression; Thrombosis