• Proceedings of the Microbiological Society of Korea Conference
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• 1999.04a
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• pp.59.2-59.2
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• 1999

• Journal of Life Science
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• v.19 no.10
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• pp.1478-1483
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• 2009

A solid-state culture based on grain materials was attempted to produce a fibrinolytic enzyme for blood circulation improvement using Bacillus subtilis BK-17. The spore, rather than vegetative cell inoculation, of B. subtilis BK-17 on the solid-state culture was effective in the production of a fibrinolytic enzyme. Maximum spore production was obtained with a SFM medium (0.8% nutrient broth, 0.05% yeast extract, $10^{-1}$ M $MgCl_2$, $10^{-3}$ M $FeCl_3$, $10^{-4}$ $MnCl_2$, $10^{-5}$ M dipicolic acid, pH 6.5). Optimal pH and temperature were pH 6 and $30^{\circ}C$, respectively. The spore production reached a maximum at 60 hours of incubation. Bacillus subtilis BK-17 on the mung bean solid-state culture produced greater fibrinolytic activity, and less activity was seen in other grains such as kidney bean, soybean and corn. Protein and lipid contents of fermented soybeans were about 10 - 30% more than those of unfermented soybeans. Amino acid content was also 5 - 20% more than that of unfermented soybeans. These results indicated that fermented solid-state culture medium, fermented soybean in this case, can be utilized as a food supplement.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.434-438
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• 2005

In the previous study, we isolated a myulchikinase (MK), which has fibrinolytic activity and cytotoxicity to the tumor cell line, from myl- chi-jeot-gal. In this study, the effect of NaCl concentration, metallic ions, pH, temperature, and plasminogen on the activity of MK was analysed. The MK activity was maintained at least $80\%$ activity up to $30\%$ NaCl, which indicates that the enzyme may be halotolerant. The optimal pH and temperature were 8 and $40^{\circ}C$, respectively. The fibrinolytic activity of MK was completely inhibited with 0.5 mM $Hg^{2+}$ and inhibited to $50^{\circ}C$ with 1 mM $Cu^{2+}\;and\;Zn^{2+}$. The MK showed strong activity in plasminogen- rich fibrin plate but not in plasminogen-free fibrin plate. The result indicates that the MK may be a plasminogen activator type fibrinolytic enzyme.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.1
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• pp.41-46
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• 2004

A correlation between fibrinolytic activity and microflora in Korean traditional soybean fermented food was investigated. The fibrinolytic activities of traditional soybean pastes and commercially processed samples were 2.42$\pm$1.01 unit/g and 1.58$\pm$0.98 unit/g, respectively. The cell density of Bacillus in traditional soybean pastes was about 10$^{7}$ CFU/g and its commercially processed one was 10$^{6}$ CFU/g. Acid producing bacteria, fungi and yeast group were higher in commercially processed one. The correlations of fibrinolytic activity and microflora in traditional and commercial Doenjang were positively correlated in Bacillus ($R^2$≒ 0.69), negatively correlated in fungal group ($R^2$≒0.40), and there were no significant correlations in acid forming bacteria and yeast group ($R^2$<0.16). Fibrinolytic activities in Meju and Koji were 6.54$\pm$1.97 unit/g and 1.46$\pm$0.43 unit/g respectively, and were positively correlated with Bacillus. Yeast and acid forming bacteria were grown by 5∼6 decimal induction during fermentation period of Doenjang, but Bacillus, fungal cells and fibrinolytic activity were nearly stable. Results indicate that fibrinolytic activity of Doenjang depends on enzyme induction in Meju or Koji processing by Bacillus, Doenjang fermentation process.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.347-356
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• 2019

Bacillus sp. BS2 showing strong fibrinolytic activity was isolated from sea squirt (munggae) jeotgal, a traditional Korean fermented seafood. BS2 was identified as B. velezensis by molecular biological methods. B. velezensis BS2 grows well at 15% NaCl and at $10^{\circ}C$. When B. velezensis BS2 was cultivated in TSB broth for 96 h at $37^{\circ}C$, the culture showed the highest fibrinolytic activity ($131.15mU/{\mu}l$) at 96 h. Three bands of 27, 35 and 60 kDa were observed from culture supernatant by SDS-PAGE, and fibrin zymography showed that the major fibrinolytic protein was the 27 kDa band. The gene (aprEBS2) encoding the major fibrinolytic protein was cloned, and overexpressed in heterologous hosts, B. subtilis WB600 and E. coli BL21 (DE3). B. subtilis transformant showed 1.5-fold higher fibrinolytic activity than B. velezensis BS2. Overproduced AprEBS2 in E. coli was purified by affinity chromatography. The optimum pH and temperature were pH 8.0 and $37^{\circ}C$, respectively. $K_m$ and $V_{max}$ were 0.15 mM and $39.68{\mu}M/l/min$, respectively, when N-succinyl-Ala-Ala-Pro-Phe-pNA was used as the substrate. AprEBS2 has strong ${\alpha}$-fibrinogenase and moderate ${\beta}$-fibrinogenase activity. Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at $10^{\circ}C$, B. velezensis BS2 can be used as a starter for jeotgal.

• Proceedings of the KOSOMBE Conference
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• v.1998 no.11
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• pp.227-228
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• 1998

Lumbrokinase, potent fibrinolytic enzyme purified from earthworm, was immobilized onto polyurethane valves using photoreaction, photoreactive polyallyl-amino as a photoreactive linker. For evaluation of blood compatibility, lumbrokinase immobilized polymer valves were assembled into the total artificial heart (TAH). This TAH was implanted to 60kg healthy lamb for 1-3 days with the cardiac output 5 L/min. In the control lamb, the valves were untreated, in ore other, only valves on the right were treated, and in the remaining animal, only those on the left. To facilitate the thrombus formation, low doses of heparin were administered. For evaluation of the immobilized lumbrokinase, thrombus formation, proteolytic and fibrinolytic activity was measured. This data shows that lumbrokinase-treated polyurethane valves lead to decreased thrombus formation in vivo, and that their biocompatibility is therefore higher than that of untreated valves.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.370-374
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• 2010

A gene encoding the major secreted fibrinolytic protein of Bacillus amyloliquefaciens CH86-1 was cloned from genomic DNAs. DNA sequencing showed that the gene, aprE86-1, could direct the synthesis of a mature protein 275 amino acids in length after processing. When aprE86-1 was introduced into B. subtilis, a mature 27-kDa protein was produced as expected. The fibrinolytic activity of the B. subtilis transformant (TF) was higher than that of B. amyloliquefaciens CH86-1, showing the possibility of increasing the fibrinolytic activity of Bacillus strains through genetic engineering.

• Proceedings of the Korea Environmental Mutagen Society Conference
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• 2004.05a
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• pp.129-129
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• 2004

• Journal of Life Science
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• v.11 no.4
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• pp.304-310
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• 2001

In order to produce a high quality and functional black bean Chungkugjang, a bacterium which has potent enzyme activities(protease:124.8 U/$m\ell$, $\alpha$-amylase: 78.2U/$m\ell$, glucoamylase; 13.9U/$m\ell$, ), was isolated by using the halo zone method and identified by morphological, cultural and biochemical properties, and then finally confirmed by GP-microplate identification system as Bacillus megatrium SMY-212. The isolated SMY-212 system was also high in the formation of mucoid material and fibrinolytic activity.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.997-1004
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• 2009

Bacillus amyloliquefancies CH51 isolated from cheonggukjang, a traditional Korean fermented soy food, has strong fibrinolytic activity and produces several fibrinolytic enzymes. Among four different growth media, tryptic soy broth was the best in terms of supporting cell growth and fibrinolytic activity of this strain. A protein with fibrinolytic activity was partially purified from the culture supernatant by CM-Sephadex and Phenyl Sepharose column chromatographies. Tandem mass spectrometric analysis showed that this protein is a homolog of AprE from B. subtilis and it was accordingly named AprE51. The optimum pH and temperature for partially purified AprE51 activity were 6.0 and $45^{\circ}C$, respectively. A gene encoding AprE51, aprE51, was cloned from B. amyloliquefaciens CH51 genomic DNA. The aprE51 gene was overexpressed in heterologous B. subtilis strains deficient in fibrinolytic activity using an E. coli-Bacillus shuttle vector, pHY300PLK.