• Microbiology and Biotechnology Letters
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• v.26 no.6
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• pp.507-514
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• 1998

The strain K-54, the best producer of fibrinolytic enzyme, was isolated from Korean traditional food Chung Guk Jang and identified as Bacillus subtilis. Fibrinolytic enzyme was purified and characterized, and its molecular weight was determined. The fibrinolytic enzyme activity was increased about 66.9 times via purification with recovery yield of 10.1%. The optimum pH and temperature of this enzyme were 11 and $65^{\circ}C$. The enzyme was stable within a pH range 8-12 and unstable at 9$0^{\circ}C$. The molecular weight was estimated to be 29,000 dalton in the form of monomer with no other subunit. The isoelectric point was calculated 8.67. N-terminal sequence was identified Ala-Gly-Ser-Val-Pro-Try-Gly-Ser. Km value of the enzyme for $\alpha$-casein was calculated to be 0.31 (3.1 mg/$m\ell$). The enzyme activity highly inhibited by PMSF at 1 nM.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.10
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• pp.1456-1460
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• 2006

In order to determine the causes of sweetened adzuki ann spoilage, the characteristics of microorganism isolated from spoiled adzuki ann were investigated. The isolated microorganism was gram-positive, roil-shaped and shore-forming bacteria; its surface was smooth and glazed. From the results of the assimilation test of 46 different biochemicals by the Vitec 2 Compact test and comparison of the cellular wall composition of fatty acid by the data bank of Midi sherlock system, the microorganism was identified as Bacillus subtilis, D-value of the B. subtilis spore was 4.85 min at $115^{\circ}C$, 0.69 min at $121^{\circ}C$ and 0.48 min at $125^{\circ}C$; Z-value was 9.71. The Bacillus subtilis growth was not observed below water activity of 0.92 at $45^{\circ}C$. However, bacteria growth increased gradually as water activity increased above 0.93.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.6
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• pp.1039-1043
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• 1997

Correlation among color intensity, electron donating ability to $\alpha$, $\alpha$-diphenyl-$\beta$-dicrylhydrazy(DPPH) and cultivation conditions by Bacillus subtilis DC-2 were tested with response surface methodology. Both of pigment generation ability and DPPH were more affected by temperature than any other factor. The highest correlation was appeared between color intensity and DPPH as 0.8364 which is significant at 1% level. After fixing cultivation time which is not significant at 10% level to 84hrs as optical cultivation time, response surface methodology was conducted in regarding temperature and color intensity. As a result of overlapped contour map of color intensity and DPPH, when cultivation temperature was in the range of 38.9~41.1$^{\circ}C$ and pH was in the range of 8.34~9.12, optical density of color intensity was predicted higher than 0.374 at 390nm and DPPH was infered higher than 1.310 at 528nm. In the range of optical culture condition, cultivation temperature, pH and cultivation time was fixed to 4$0^{\circ}C$, 8.5 and 85hrs, respectively. In resulting, observation value of color intensity and DPPH was in the range of anticipation value as 0.386 at 390nm and 1.332 at 528nm respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.6
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• pp.915-921
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• 1996

An alkaline pretense Producing microorganism was isolated from Korean traditional soy sauce and identified as Bacillus subtilis CCKS-111. The optimum culture condition of Bacillus subtilis CCKS-111 for the production of alkaline pretense was as follow: 2% soluble starch, 0.2% peptone, 0.1% (NB$_4$)$_2$S$_2$O$_{8}$ , 0.2% MgSO$_4$, pH 7.0, 35$^{\circ}C$ and 24hrs. The optimum pH and temperature for the enzyme activity of alkaline pretense producing Bacillus subtilis CCKS-111 were pH 9.0 and 5$0^{\circ}C$, respectively. The enzyme was relatively stable at pH 6.0~11.0 and at temperature below 4$0^{\circ}C$. The activity of the enzyme was inhibited by $K^{+}$ and Hg$^{2+}$, whereas Cu$^{2+}$ exhibited rather activating effects on the enzyme activity. Ethylenediaminetetraacetic acid and phenylmethanesulfonyl fluoride inhibited the enzyme activity. This indicates that this is serine pretense which requires metal ion group for the enzyme activity. Km value was 2.313$\times$10$^{-4}$ M/L, V$_{max}$ value was 39.216$\mu\textrm{g}$/min. This enzyme hydrolyzed casein more rapidly than the hemoglobin.lobin.

• KSBB Journal
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• v.13 no.5
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• pp.491-496
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• 1998

The production of fibrinolytic enzyme from Bacillus subtilis BK-17 was studied in the shake flask cultures. The important medium components studied include nitrogen source, carbon source and inorganic salts. The environmental conditions include initial pH, temperature, shaking speed and working volume. Among various N-sources, C-sources and inorganic salts tested, soybean flour, D-glucose and Na2HPO4 gave the best results, and their optimal concentrations were 1.5%, 0.5% and 0.05%, respectively. The optimal pH and temperature were 9.0 and 37$^{\circ}C$. With decreasing working volume in the range of 25∼100ml in the 250ml flask or increasing shaking speed in the range of 100∼300rpm, the enzyme production was greatly enhanced. The enzyme activity under the optimal conditions was about 1400I.U./ml with urokinase as a standard.

• The Korean Journal of Pesticide Science
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• v.16 no.2
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• pp.178-186
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• 2012

This study was investigated for the purpose of the isolation and identification of antagonistic bacteria with antifungal activity against plant pathogens. This bacteria denominated Bacillus subtilis KYS-10 and the optimum growth condition were 4% sucrose, 1% yeast extract, 0.2% $K_2HPO_4$, pH 7, 150 rpm, $30^{\circ}C$, 8 day. The antifungal activities against nine plant pathogens determined inhibition zone size by diffusion methods. The results, G. zeae (scab) 70 mm and P. grisea KACC 40439 (blast), P. capsici KACC 40177 (phytophthora blight) and C. destructans KACC 41077 (root rot of ginseng) 40~43 mm, and C. gloeosporioides KACC 43520 (ripe rot), C. gloesporioides KACC 40003 (anthracnose), S. shiraiana KACC 41065 (stem rot) and S. shiraiana (mulberry sclerotial disease) 35~39 mm and F. Oxysporum KACC 44452 (bulb rot of ginseng) 28 mm. From these experiment results, author suggest that Bacillus subtilis KYS-10 would be developed as a biological control agent thorough the field experimet in the near future.

• KSBB Journal
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• v.25 no.5
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• pp.429-436
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• 2010

Keratinase is widely used in certain industrial applications. The present study sought to improve the culture conditions of Bacillus subtilis SMMJ-2 to facilitate mass production of keratinase. Strain SMMJ-2 was irradiated by ultraviolet light and the resulting isolates were tested for keratinase activity. Isolates displaying elevated keratinase activity were selected and used to determine the optimum temperature (24, 30, 37, 45, $55^{\circ}C$) for bacterial keratinase production during a 4 day incubation period. The highest enzyme activity (55 units/mL/min), from a Bacillus subtilis SMMJ-2 mutant (mutant No. 2) was demonstrated following incubation at $30^{\circ}C$. The effects of carbon and nitrogen sources on keratinase production were confirmed by measuring the enzyme activity from the culture broth of the mutant strain cultured in various media containing different carbon source and nitrogen sources during a 4 day period. The optimal medium composition for producing keratinase consisted of 1% glucose, 0.7% $K_2HPO_4$, 0.2% $K_2HPO_4$, and 1.2% soybean meal. Optimal initial pH and temperature for producing keratinase were 7.0 and $30^{\circ}C$, respectively. Keratinases produced by B. subtilis SMMJ-2 and the mutant No. 2 were purified from the culture broth which used soybean meal as a nitrogen source. Membrane ultrafiltration, DEAE-sephacel ion exchange and Sephadex G-100 gel chromatography were used to purify the enzymes. The purified keratinases from both B. subtilis SMMJ-2 and the mutant No. 2 showed single bands and their molecular weights were estimated as 28 kDa and 42 kDa, respectively on SDS-polyacrylamide gel electrophoresis.

• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1092-1100
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• 2012

Ten Bacillus strains with antimicrobial activities were isolated from Cheonggukjang produced at different parts in Korea. They all inhibited Listeria monocytogenes ATCC 19111 and nine inhibited Bacillus cereus ATCC 14579. Four isolates (W42, H27, SKE 12, and K21) showing strong inhibiting activities were identified as B. subtilis. B. subtilis W42 was the most inhibiting strain. The antimicrobial activity of culture supernatant from B. subtilis W42 was destroyed completely by proteinase K treatment, indicating that a bacteriocin was the responsible agent. The bacteriocin, Bac W42, was most stable at pH 7 and stable between pH 3-6 and 8-9. Bac W42 was stable up to $80^{\circ}C$. BHI (brain heart infusion) and TSB (tryptic soy broth) were the best media for the activity (320 AU/ml) followed by LB (160 AU/ml). Bac W42 was partially purified by column chromatographies. The specific activity was increased from 1,151.2 AU/ml to 9,043.5 AU/ml and the final yield was 26.3%. Bac W42 was 5.4 kDa in size as determined by SDS-PAGE. Bac W42 showed bactericidal activity against L. monocytogenes ATCC 19111.

• Applied Biological Chemistry
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• v.39 no.6
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• pp.460-465
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• 1996

The production of bacterial protease and its characteristics were investigated with Bacillus subtilis globigii CCKS-118 which was isolated from Korean traditional soy sauce. The optimum culture condition of the strain for the production of alkaline protease was as follow : 2% soluble starch, 0.2% yeast extract, 0.1% $(NH_4)_2SO_4$, 0.2% $MgSO_4$, pH 7.5, $35^{\circ}C$ and 20h rs. The optimum pH and temperature for the enzyme action of alkaline protease producing Bacillus subtilis globigii CCKS-118 were pH 9.0 and $50^{\circ}C$, respectively. The enzyme was relatively stable at $pH\;6.0{\sim}9.0$ and at temperature below $40^{\circ}C$. The activity of the enzyme was inhibited by $Hg^{2+}$ whereas $Cu^{2+}$ gave rather activating effects on the enzyme activity. The enzyme was inhibited by phenylmethane-sulfonyl fluoride indicating serine pretense metal ion group are required for the enzyme activity. Km value was $1.242{\times}10^{-4}M$, $V_{max}$ value was $25.99\;{\mu}g/min$. This enzyme hydrolyzed casein more rapidly than the hemoglobin.

• Korean Journal of Microbiology
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• v.17 no.2
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• pp.72-80
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• 1979

24 samples which were incoulated with Aspergillus flavus and Bcillus subtilis and cultured on the steamed soybean media under various conditions-pH, moisture and temperature were-investigated on the production of aflatoxin by the interaction of these two microorganisms. 1) The amount of aflatoxin produced by mixed cultures of Aspergillus flavus and Bacillus subtilis was decreased significantly rather than that of single cultures of Aspergillus flavus. 2) Maximum production of crude aflatoxin was 2,560 ppm $(B_1,\;0.908\;ppm;\;B_2,\;0.261\;ppm;\;G_1,\;1.162\;ppm;\;G_2,\;0.229\;ppm)$ at 30% moisture, pH 5.0 and $20^{\circ}C$, whereas minimum production was 1.107 ppm $(B_1,\;0.341\;ppm;\;B_2,\;0.104\;ppm;\;G_1,\;532\;ppm;\;G_2,\;0.130\;ppm)$ at 63% moisture, pH 9.0 and $40^{\circ}C$.