• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.185-193
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• 2016

A novel proteolytic bacterium was isolated from soil at Yeungnam University, South Korea. The strain, named FBL-1, was rod-shaped with a smooth surface. Biolog and API 50CHB test results revealed that strain FBL-1 was a Bacillus species. Based on 16S rDNA sequencing and chemotaxonomic characterization, the strain was identified as Bacillus subtilis because it had the highest homology with Bacillus subtilis subsp. subtilis NCIB 3610 (99.5%). In liquid culture at 37℃ with shaking at 200 rpm, fructose and yeast extract were found to be the best carbon and nitrogen sources, respectively, for cell growth and protease production. The highest protease activity (451.640 U/ml) was obtained when the strain was cultured in medium containing 20 g/l of fructose and 5 g/l of yeast extract. Although further studies are needed to characterize the protease and enhance its activity, the newly isolated protein-degrading B. subtilis FBL-1 can be applicable for the production of peptides and for the degradation of proteins in various industries.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.283-289
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• 2002

This study was performed to search for potential microorganism that has rapid fermenting and physiological function from anchovy sauce. We isolated three bacterial strains, JM-1, JM-2, and JM-3 with proteolytic and fibrinolytic activity from anchovy sauce. Among the 3 bacterial strains, JM-3 showed the strongest proteolytic and fibrinolytic activity. Bacterial strain JM-3 was gram-positive rod, motile and formed endospore. The 16S rRNA of bacterial strain JM-3 was amplified by PCR and then its sequence was determined by ABI 310 genetic analyzer. The 16S rRNA sequence of bacterial strain JM-3 was compared to BLAST DNA database and identified to Bacillus subtilis with 99% of homology. The optimum temperature, pH and NaCl concentration for growth of B. subtilis JM-3 were $40^{\circ}C$, 5.0 and 0%, respectively. The optimum temperature, pH and NaCl concentration for proteolytic and fibrinolytic enzyme production of B. subtilis JM-3 were same as optimum conditions for growth. At 20% of NaCl concentration which is common NaCl concentration of fish sauce, B. subtilis JM-3 showed about 60% of proteolytic and fibrinolytic activity of 0% NaCl concentration. From above results, we found that B. subtilis JM-3 will be able to used for starter of functional fish sauce.

• Korean Journal of Environmental Biology
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• v.22 no.4
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• pp.487-493
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• 2004

To investigate the antifungal activity related protein in pesticidal bacteria, a bacterial strain LTD was isolated from soil collected at Gimje in Jeonbuk province, Korea, and identified as Bacillus subtilis LTD based on a API50 CHB kit and 168 rDNA sequencing. It has an antifungal activity against 9 plant pathogenic fungi in a paper disc assay. The antifungal activity- deficient mutant, B. subtilis mLTD was induced at a 5 kGy dose of $^{60}Co$ gamma radiation. Using the two-dimensional electrophoresis and the matrix assisted laser desorption ionization time-of-flight mass spectrometry, the comparison analysis of proteins between the wild and mutant were performed. A major intracellular serine proteinase IspA (MW: 32.5 kDa), a NAD (P) H dehydrogenase (MW: 20.0 kDa), and a stage II sporulation protein AA, SpoIIAA (MW: 14.3kDa) were detected only in the B. subtilis LTD. These results suggested that the functions of these proteins found only in the B. subtilis LTD could. be closely related to the antifungal activity against plant pathogenic fungi.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.512.2-512
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• 1986

A mutant of bacillus subtilis with a defective cdd gene encoding deoxycytidine-cytidine deaminase(EC 3.5.4.5.) has been characterized genetically. The genetic lesion causing the altered deoxycytidine-cytidine deaminase, cdd, was mapped at 225 min on the linkage map of B.subtilis by AR9 transduction Transductional analysis of the cdd region established the gene order as trp-lys-dnaE-cdd-aroD. The cdd gene was linked 72% with the aroD and 20% with the lys.

• Korean Journal of Microbiology
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• v.45 no.2
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• pp.185-192
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• 2009

Chungkookjang fermenting Bacillus subtilis 028-1 strain suppressed the growth of Staphylococcus sp. LS2, Saccharomyces cerevisiae, and Candida albicans. B. subtilis 028-1 strain produced antibiotic effectively in the medium of 2% soybean meal and 1% maltose as a disaccharide, when the shaking was continued 15~18 h and the pH of culture medium was maintained under 6.5. The antibiotic activity was optimized when the initial pH of the culture medium of test strain was adjusted with weak alkali, was remained after 20 min of boiling and for more than 1 month in room temperature, and was weakened slowly by the digestion of chymotrypsin and papain. The molecular weight of the antibiotic was identified between 500 and 1,000 dalton by dialysis, and antibiotic substance was considered as not surfactin but a member of iturin family because of the absence of fibrinolytic activity.

• The Korean Journal of Community Living Science
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• v.18 no.3
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• pp.399-406
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• 2007

Soybean seeds were fermented by Bacillus subtilis and/or Lactobacillus bulgaricus to improve solubility, viscosity, water holding capacity and oil holding capacity of soybean proteins in Chongkukjang. The maximum colony forming unit and protease activity of B. subtilis or L bulgaricus were observed after 60 hours of fermentation, and those of the mixed fermentation by two microorganisms were steadily increased during the fermentation periods. Solubilities of soybean proteins by B. subtilis or L bulgaricus were steadily increased before the values were considerably increased to 60 hours of fermentation, whereas water holding capacities of the proteins were decreased by B. subtilis or L. bulgaricus and those of the mixed fermentation were decreased progressively. Viscosities of soybean proteins by B. subtilis and/or L. bulgaricus were decreased progressively during the fermentation. Viscosities of soybean proteins by B. subtilis and/or L. bulgaricus were decreased progressively during the fermentation. Oil holding capacities of soybeans by B. subtilis or L. bulgaricus were maximum at 20 or 80 hours of fermentation and those of the mixed fermentation were decreased after 10 hours of the fermentation.

• Applied Biological Chemistry
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• v.46 no.2
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• pp.69-73
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• 2003

A novel antifungal antibiotic for azole-resistant Candida albicans was purified from the culture broth of Bacillus subtilis LAM 97-44 by butanol extraction, Diaion HP-20 and Dowex-50 adsorption chromatography, silica gel flash chromatography followed by HPLC and designated LAM-44A. LAM-44A was stable for 60 min at $100^{\circ}C$, and pH range from 2 to 10. MIC values were observed at $0.5-3.5\;{\mu}g/ml$ against various Candida albicans strains. The antibiotic showed no cytotoxicity for S180, MKN-45, P388, HeLa and 373 at the concentration of 1 mg/ml. LAM-f4A was colorless powder soluble in water, methanol, ethanol, butanol and negative to ninhydrin reaction. The antibiotic had maximum absorption at 273 nm in methanol, and melting point was $202^{\circ}C$. The molecular weight and formula were determined to be 282 and $C_{14}H_{34}O_5$ by $^1H-NMR,\;^{13}C-NMR$, IR spectrum and elemental analysis.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.8
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• pp.1120-1127
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• 2011

A feeding trial was conducted to investigate the effect of dietary supplementation of Bacillus subtilis LS 1-2 grown on citrus-juice waste and corn-soybean substrate on growth performance, nutrient retention, caecal microbial population and intestinal morphology in broilers. Three hundred twenty d-old Ross chicks were randomly allotted to 4 treatments on the basis of BW in a randomized complete block design. Each treatment had 4 replicates of 20 chicks in each. Experimental diets were fed in 2 phases, starter (d 0 to 21) and finisher (d 21 to 35). Dietary treatments were; negative control (NC: basal diet without any antimicrobial), positive control (PC: basal diet added with 20 mg/kg Avilamycin), basal diet added with 0.30% Bacillus subtilis LS 1-2 grown on corn-soybean substrate (P1), and basal diet added with 0.30% Bacillus subtilis LS 1-2 grown on citrus-juice waste substrate (P2). Overall BW gain, feed intake and FCR were better (p<0.05) in PC, P1 and P2 treatments as compared to NC. Moreover, overall BW gain and FCR in PC and P2 treatments were greater than P1. Retention of CP, GE (d 21, d 35) and DM (d 35) were increased (p<0.05) in treatments PC, P1 and P2 compared with NC. At d 35, caecal Clostridium and Coliform counts were lower (p<0.05) in treatments PC, P1 and P2 than NC. Moreover, Clostridium and Coliform counts in treatment PC was lower (p<0.05) than P1. Villus height and villus height to crypt depth ratio in both duodenum and ileum were increased (p<0.05) in treatments PC, P1, P2 as compared to NC. However, retention of nutrients, caecal microbial population and intestinal morphology remained comparable among treatments P1 and P2. It is concluded that Bacillus subtilis LS 1-2 inclusion at 0.30% level had beneficial effects on broilers' growth performance, nutrient retention, caecal microflora and intestinal morphology. Additionally, citrus-juice waste can be used as substrate for growth of probiotic Bacillus subtilis LS 1-2.

• Korean Journal of Microbiology
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• v.43 no.4
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• pp.331-336
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• 2007

A bacterium producing the alkaline pretense was isolated from Chungkookjoug, and was identified as Bacillus subtilis JK-1 based on morphological, physiological and biochemical characteristics, as well as phylogenetic analysis using 16S rRNA gene sequence. The optimum pH and temperature of the pretense activity were pH 9.0 and $55^{\circ}C$, respectively. This enzyme was stable at the temperatures $40{\sim}80^{\circ}C$. The maximum alkaline pretense production was obtained when 1.0% (w/v) xylose, 1.0% (w/v) yeast extract and 0.3% (w/v) $CuSO_4$ were used as carbon source, nitrogen source and mineral source. Under the optimal condition, growth of the isolate was reached at stationary phase after 12 hr followed by incubation, the alkaline pretense production reached a maximum level with $16{\sim}20$ hr cultivation.