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Research article

J. Microbiol. Biotechnol. 2015; 25(12): 2090-2099

Published online December 28, 2015 https://doi.org/10.4014/jmb.1509.09048

Copyright © The Korean Society for Microbiology and Biotechnology.

Characterization of a Novel Fibrinolytic Enzyme, BsfA, from Bacillus subtilis ZA400 in Kimchi Reveals Its Pertinence to Thrombosis Treatment

Min-Ju Ahn 1, Hye-Jin Ku 1, Se-Hui Lee 1 and Ju-Hoon Lee 1*

Department of Food Science and Biotechnology, Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Republic of Korea

Received: September 16, 2015; Accepted: September 21, 2015

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 (4-40oC), metal ions (Mn2+, Ca2+, K2+, and Mg2+), 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.

Keywords: Thrombosis, Fermented food, Bacillus subtilis, Fibrinolytic activity, Gene expression