• Microbiology and Biotechnology Letters
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• v.17 no.2
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• pp.160-164
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• 1989

A 5.7Kb EcoRI fragment containing alkaline amylase gene of Bacillus sp. AL-8 obtained in the previons experiment (10) was transformed in B. subtilis via plasmid pUB110. The enzymatic proper-ties of the amylase produced by the transformants were Identical to those of the donor strain. Thus, the alkaline amylase activity from the transformant was maximum at pH 10 and 5$0^{\circ}C$. And the enzyme was very stable over the ranges of alkaline pH. In order to determine the location of the alkaline amylase gene within the 5.7Kb DNA fragment, the fragment was subcloned in E. coli. It was found that the alkaline amylase gene was located k EcoRI fragment of 3.7Kb.

• Journal of Food Hygiene and Safety
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• v.15 no.2
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• pp.122-127
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• 2000

An antifungal bacterium was isolated to inhibit of the growth of Asp. flavus and Asp. parasiticus, and its antifungal compounds were purified from lyophilized culture broth using chromatographic methods. Antifungal bacterium #19 which was shown a higher inhibitory activity on the growth of aflatoxin producing fungi was identified as Bacillus subtilis. The purified antifungal compound(1 mg) was demonstrated strong antifungal activity against the aflatoxin producing fungi.

• Food Science and Biotechnology
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• v.17 no.6
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• pp.1372-1375
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• 2008

The aprE2 gene from Bacillus subtilis CH3-5 was expressed in Bacillus licheniformis ATCC 10716 using a Bacillus-Escherichai coli shuttle vector, pHY300PLK. The fibrinolytic activity of transformant (TF) increased significantly compared to B. licheniformis 10716 control cell. During the 100 hr incubation in Luria-Bertaini broth at $37^{\circ}C$, fibrinolytic activity of B. licheniformis TF increased rapidly at the late growth stage, after 52 hr of incubation, which was confirmed by zymography using a fibrin gel. pHY3-5 was stably maintained in B. licheniformis without tetracycline (Tc) in the media, 60.9% of cells still maintained pHY3-5 after 100 hr of cultivation.

• Research in Plant Disease
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• v.12 no.2
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• pp.85-90
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• 2006

Bacillus subtilis BD0310 isolated from tea leaves was used for the development of a biofungicide against Pestalotiopsis longiseta causing gray blight of tea plants. The optimum growth conditions were investigated for the mass cultivation of the microbial agent. The optimum temperature and cultivation time were determined as $12{\sim}24$ hours at $30^{\circ}C$ and the optimum initial pH was pH 7.0 in nutrient broth. Among the tested carbon sources of fructose, galactose, glucose, glycerol, inositol, lactose, maltose, sorbitol and starch, maltose and inositol were found to highly increase antifungal activity of the microbial agent against P. longiseta. Yeast extract and tryptone apparently increased antifungal activity of the microbial agent among the tested nitrogen sources of casein, tryptone, malt extract, yeast extract and $(NH_4)_2SO_4$. The results will make a contribution to mass production of the antagonistic bacterium Bacillus subtilis BD0310 for development of a microbial agent controlling gray blight of tea plants.

• 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.

• Fashion & Textile Research Journal
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• v.8 no.2
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• pp.239-244
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• 2006

For studies of fibrinolytic enzyme strain K-54 was isolated from the Korean traditional food chungkook-jang. Isolated strains K-54 was identified as Bacillus subtilis. The molecular weight of fibrinolytic enzyme from B. subtilis K-54 was 27 kDa. Optimum temperature for fibrinolytic enzyme of B. subtilis K-54 was $50-70^{\circ}C$ and optimum pH for producing the enzyme of this strain was ranging from 8 to 12. Also, it was found out enzyme activity was completely inhibited by 1mM PMSF. The result indicated this enzyme was thermo-stable alkaline serine protease with strong fibrinolytic activity. The wool and silk were treated with protease of B. subtilis K-54. As a result, the property of dyeing of wool fabrics was increased. By the increasing of treatment time became smoothened. But the change of mechanical properties were not changed.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.128-137
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• 2023

Transcriptome analysis revealed that the sinR gene encoding a transition-state regulator of Bacillus pumilus, genetically close to B. subtilis, was expressed at high levels during growth. The sinR gene is the second gene of the sinIR operon consisting of three promoters and two structural genes in B. subtilis. This study used the sinIR promoter of B. subtilis DB104 to construct a recombinant protein expression system. First, the expression ability depending on the number of sinIR promoter was investigated using enhanced green fluorescent protein (eGFP). The expression level of eGFP was slightly higher when using two promoters (Psin2) than using original promoters. The Psin2 promoter was further engineered by modifying the repressor binding site and -35 and -10 regions. Shine-Dalgarno (SD) sequence of the sinI gene was modified to the consensus sequence. Finally, combining the engineered Psin2 promoter with the modified SD sequence increased the expression level of eGFP by about 13.4-fold over the original promoter. Our results suggest that the optimized sinIR promoter could be used as a novel tool for recombinant protein expression in B. subtilis.

• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.122-129
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• 1998

It were reported that antifungal mechanism of Enterobacter cloacae is a volatile ammonia that produced by the strain in soil, and the production of ammonia is related to the bacterial urease activity. A powerful bacterium SH14 against soil-borne pathogen Fusarium solani, which cause root rot of many important crops, was selected from a ginseng pathogen suppressive soil. The strain SH14 was identified as Bacillus subtilis by cultural, biochemical, morphological method, and $API^{circledR}$ test. From several in vitro tests, the antifungal substance that is produced from B. subtilis SH14 was revealed as heat-stable and low-molecular weight antibiotic substance. In order to construct the multifunctional biocontrol agent, the urease gene of Bacillus pasteurii which can produce pathogenes-suppressive ammonia transferred into antifungal bacterium. First, a partial BamH I digestion fragment of plasmid pBU11 containing the alkalophilic B. pasteurii l1859 urease gene was inserted into the BamH I site of pEB203 and expressed in Escherichia coli JM109. The recombinant plasmid was designated as pGU366. The plasmid pGU366 containing urease gene was introduced into the B. subtilis SH14 with PEG-induced protoplast transformation (PIP) method. The urease gene was very stably expressed in the transformant of B. subtilis SH14. Also, the optimal conditions for transformation were established and the highest transformation frequency was obtained by treatment of lysozyme for 90 min, and then addition of 1.5 ${mu}g$/ml DNA and 40% PEG4000. From the in vitro antifungal test against F. solani, antifungal activity of B. subtilis SH14(pGu366) containing urease gene was much higher than that of the host strain. Genetical development of B. subtilis SH14 by transfer of urease gene can be responsible for enhanced biocontrol efficacy with its antibiotic action.

• The Korean Journal of Food And Nutrition
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• v.21 no.1
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• pp.15-21
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• 2008

In this study, the minimum inhibition concentration(MIC), growth inhibition activity, and colony forming inhibitory activity of water soluble propolis against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Streptococcus mutans, Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae and Salmonella enteritidis were tested. The MICs of the water soluble propolis against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Streptococcus mutans, Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae, and Salmonella enteritidis were 312.5 ppm, below 156.3 ppm, 625 ppm, 10,000 ppm, above 10,000 ppm, 10,000 ppm, above 10,000 ppm, above 10,000 ppm, 10,000 ppm, and above 10,000 ppm, respectively. The growth inhibition concentrations against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Streptococcus mutans, Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, and Klebsiella pneumoniae were 156.3 ppm, below 156.3 ppm, 625 ppm, 5,000 ppm, 10,000 ppm, 10,000 ppm, 10,000 ppm, 10,000 ppm, and 5,000 ppm, respectively. However, 10,000 ppm did not inhibit the growth of Salmonella enteritidis. Finally, the colony forming inhibitory activities against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Streptococcus mutans, Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae, and Salmonella enteritidis were 98.0%, 99.8%, 69.8%, 98.1%, 62.0%, 63.1%, 79.5%, 61.9%, 79.6%, and 0.0%, respectively.

• 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.