• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.9
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• pp.1114-1121
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• 2017

Bacillus strains not producing harmful components were isolated from Korean traditional soybean products. Extracellular enzyme activities (amylase, protease, cellulase, and xylanase) of isolated Bacillus strains were measured, and Bacillus strains with high protease activity were selected. The selected 15 strains were identified as Bacillus amyloliquefaciens (10), Bacillus methylotrophicus (1), Bacillus velezensis (1), and Bacillus subtilis (3). Among them, B. subtilis JBG17019, B. amyloliquefaciens JBD17076, and B. amyloliquefaciens JBD17109 showed antimicrobial activities against food-borne microorganisms. The production abilities of glutamate, glutamine, and poly-${\gamma}$-glutamic acid (${\gamma}$-PGA) of the selected Bacillus strains were measured to analyze fermentation characteristics related to glutamic acid metabolism. The factor for multivariate was analyzed by the principal components analysis (PCA) method between fermentation characteristics and ${\gamma}$-PGA production. The three principal components were classified according to the PCA method: PC1 [enzyme activity (amylase, cellulase, and xylanase)], PC2 (${\gamma}$-PGA), and PC3 (protease, glutamate, and glutamine). As a result, B. amyloliquefaciens JBD17076 and B. subtilis JBG17019 strains were evaluated as having excellent enzyme activity and ${\gamma}$-PGA production.

• KSBB Journal
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• v.26 no.3
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• pp.199-205
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• 2011

Using tetrameric ${\beta}$-galactosidase as a model protein, anchoring motives were screened in Bacillus subtilis spore display system. Eleven spore coat proteins were selected considering their expression levels and the location in the spore coat layer. After chromosomal single-copy homologous integration in the amyE site of Bacillus subtilis chromosome, cotE and cotG were chosen as possible spore surface anchoring motives with their higher whole cell ${\beta}$-galactosidase activity. PAGE and Wester blot of extracted fraction of outer layer of purified spore, which express CotE-LacZ or CotG-LacZ fusion verified the existence of exact size of fusion protein and its location in outer coat layer of purified spore. ${\beta}$-galactosidase activity of spore with CotE-LacZ or CotG-LacZ fusion reached its highest value around 16~20 h of culture time in terms of whole cell and purified spore. After intensive spore purification with lysozyme treatment and renografin treatment, spore of BJH135, which expresses CotE-LacZ, retained only 1~2% of its whole cell ${\beta}$-galactosidase activity. Whereas spore of BJH136, which has cotG-lacZ cassette in the chromosome, retained 10~15% of its whole cell ${\beta}$-galactosidase activity, proving minor perturbation of CotG-LacZ, when incorporated in the spore coat layer of Bacillus subtilis compared to CotE-LacZ. Usage of Bacillus subtilis WB700, of which 7 proteases are knocked-out and thereby resulting in 99.7% decrease in protease activity of the host, did not prevent the proteolytic degradation of spore surface expressed CotG-LacZ fusion protein.

• Journal of The Korean Society of Grassland and Forage Science
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• v.18 no.3
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• pp.195-204
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• 1998

This study was designed to investigate the effects of antagonistic microorganism, Bacillus subtilis, on the growth of forage seedlings in repeated cultivation soils and unrepeated cultivation soils. The field experiment was wnducted in pots in a vinyl house using repeated and unrepeated cultivation soils. Forage types were 'Suwon 19' wrn(Zea mqs L.), 'Califbmia' white clover(Tr~oIium repens L.) and 'Fawn' tall fescue (Festuca arundianacea Schreb.). Samples of white clover and tall fescue were taken h m each pot at 36 days after seeding. Samples of wm were examined at 50 days after seeding. The most active antagonistic bacterium was isolated h m forage rhizosphere soil, and selected by reference to it's antagonistic ability on the growth of pathogenic fungi, Rhizoctonia solmi and Fusarium oxyspomm, and it was identified as Bacillus subtilis. This strain strongly suppressed the growth of fungal pathogens among isolated rhizobacteria. The dry weight of forage shoots and roots cultivated in unrepeated cultivation soils was higher than that cultivated in repeated cultivation soils. The dry weight of forage was positively affected by the inoculation of the antagonistic bacterium, Bacillus subtilis, in both repeated cultivation soils and unrepeated cultivation soils. In conclusion, the growth of forage was more affected by the inoculation of the antagonistic bacterium in unrepeated cultivation soils than that in repeated cultivation soils, and bacterization of forage with B. subtilis resulted in an inrreased yield.

• Microbiology and Biotechnology Letters
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• v.26 no.3
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• pp.226-231
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• 1998

The medium optimization was investigated to maximize the production of fibrinolytic enzyme by Bacillus subtilis KCK-7 isolated from Chungkookjang, which could hydrolyze the fibrin produced through the blood coagulation mechanism in human body. The simultaneous addition of 5% soluble starch and 0.5% cellobiose to the medium as carbon sources resulted in the highest production of the fibrinolytic enzyme. Likewise, the optimized composition of medium appeared to be 0.5% peptone, 0.3% beef extract, 0.5% cellobiose, 5% soluble starch, 2% raw soybean meal and 0.02% Na$_2$HPO$_4$. In addition, the fibrinolytic enzyme production by Bacillus subtilis KCK-7 reached to the maximum level after the cultivation for 48 hr, using the optimized medium.

• Korean Journal of Soil Science and Fertilizer
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• v.34 no.5
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• pp.333-341
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• 2001

The Compost-decomposing-bacteria was isolated from livestock compost containing sawdust. The isolated bacteria was identified as Bacillus subtilis LYH201 by the method of the composition of the fatty acid with MIDI system and Bergey's manual. Cloning of CMCase encoding gene was accompanied by shotgun method. The pLK100 have yellow activity ring on CMC medium, that was carried 2.2 kb insert DNA in pBluescript II $SK^+$ vector, named BglC gene. The BglC was very similar to Pectobacterium carotovorum Gun_CLOAB(P15704) with score of 57% identity and 71% homology over 508 aa. The BglC was measured molecular weight 56 kDa by CMC-SDS-PAGE. Optimum cellulase activity Bacillus subtilis LYH201 was temperature $50^{\circ}C$ and pH 7.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.12
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• pp.1694-1699
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• 2011

The quality characteristics and physiological effects of chunggukjang fermented naturally (NF-c), with Bacillus subtilis-SKm (BS-c), with Bacillus subtilis HJ18-4 (BH-c), and with Bacillus subtilis KCCM 42923 (BK-c) were investigated. The characteristics of fermentation were determined by protease, ${\alpha}$-amylase and ${\gamma}$-GTP activities, and additionally the amounts of amino-type and ammonia-type nitrogens. BS-c showed the highest protease, ${\alpha}$-amylase, and ${\gamma}$-GTP activities, and also amino-type nitrogen content among the four types of chunggukjang. The ammonia-type nitrogen content in BS-c was similar to that of BK-c and NF-c. BH-c showed the lowest enzyme activities and amino-type and ammonia-type nitrogen content. BS-c, BH-c, BK-c, and NF-c showed a similar overall acceptability during sensory evaluation. BS-c also showed the strongest DPPH free radical scavenging and anti-proliferative activities in HT-29 human colon carcinoma cells. These results suggested that B. subtilis-SKm was suitable to be used as a starter to enhance the quality and effects of chunggukjang.

• Journal of Life Science
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• v.22 no.7
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• pp.912-919
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• 2012

A Bacillus sp. strain producing celluase and xylanase was isolated from environmental soil with LB agar plate containing carboxymethylcellulose (CM-cellulose) and beechwood xylan stained with trypan blue as substrates, respectively. Based on the 16S rRNA gene sequence and API 50 CHL test, the strain was identified as B. subtilis and named B. subtilis NC1. The cellulase and xylanase from B. subtilis NC1 exhibited the highest activities for CM-cellulose and beechwood xylan as substrate, respectively, and both enzymes showed the maximum activity at pH 5.0 and $50^{\circ}C$. We cloned and sequenced the genes for cellulase and xylanase from genomic DNA of the B. subtilis NC1 by the shot-gun cloning method. The cloned cellulase and xylanase genes consisted of a 1,500 bp open reading frame (ORF) encoding a 499 amino acid protein with a calculated molecular mass of 55,251 Da and a 1,269 bp ORF encoding a 422 amino acid protein with a calculated molecular mass of 47,423 Da, respectively. The deduced amino acid sequences from the genes of cellulase and xylanase showed high identity with glycosyl hydrolases family (GH) 5 and 30, respectively.

• Microbiology and Biotechnology Letters
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• v.43 no.3
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• pp.204-211
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• 2015

A bacterial strain capable of hydrolyzing xylan and locust bean gum (LBG) was isolated from the Saemangeum tideland of Korea. Based on the biochemical properties and the 16S rRNA gene sequence, the isolate YB-30 was identified as Bacillus subtilis. Xylanase productivity was increased effectively when B. subtilis YB-30 was grown in the presence of wheat bran, while mannanase productivity was increased drastically when grown in the presence of konjac or LBG. Particularly, maximum mannanase and xylanase activities were detected in the culture filtrate of media containing 3.5% konjac and 1% wheat bran. Both enzyme productivities reached maximum levels in the stationary growth phase. The culture filtrate exhibited the highest activity at 60℃ and pH 6.0 for mannanase and at 55℃ and pH 5.5 for xylanase, respectively. Both enzymes were not stable at high temperatures and xylanase was less stable than mannanase. In addition, wheat bran was hydrolyzed to liberate reducing sugar to a greater extent than rice bran by the culture filtrate because the wheat bran contained more arabinoxylan than the rice bran. Hence, xylanase and mannanase produced by B. subtilis YB-30 have a potential use as feed additive enzymes.

• Microbiology and Biotechnology Letters
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• v.36 no.4
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• pp.326-335
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• 2008

The siderophore ($siderophore_{AH18}$) of Bacillus subtilis AR18 was determined to be one of catechol type and purified by using Amberlite XAD-2, Sephadex LR-20 chromatography, and reversed-phase RPLC. The $Siderophore_{AH18}$ was identified bacillibactin with its structure by GC-MS, $^1H$-NMR, and $^{13}C$-NMR. $Siderophore_{AH18}$ (bacillibactin) had been confirmed its molecular weight of 883 and chemical structure of $(2,3-dihydroxybenzoate-glycine-threonine)_3$. Purified $siderophore_{AH18}$ showed strong biocontrol ability towards the spore of Phytophthora capsici on PDA and able to effectively suppress (55%) P. capsici causing red-pepper blight in the pot in vivo test.

• Journal of Applied Biological Chemistry
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• v.54 no.3
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• pp.153-158
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• 2011

Optimal culture conditions were characterized for production of crude biosurfactant of Bacillus subtilis JK-1. During incubation of B. subtilis JK-1, the bacterial growth pattern, changes of the surface tension at variable temperatures, pH and NaCl concentrations in bacterial culture medium were studied. The strain was able to grow and produce biosurfactant at $15-45^{\circ}C$, in the pH range of 6-10, and at 0-10% (w/v) NaCl. In case, culture broth pH was gradually changed to neutral or weak alkaline. Optimal culture conditions for crude biosurfactant production were at $35^{\circ}C$ and pH 7.0 after 48 h incubation and the surface tension of biosurfactant was 24.0 mN/m. Besides, as the concentration of NaCl was increased from 0 to 10% (w/v), the growth was decreased, pH of the culture broth was converted from weak alkaline to acidic, and the surface tension rised.