• Journal of Microbiology and Biotechnology
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• v.7 no.4
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• pp.223-228
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• 1997

A DNA fragment containing the gene for cell wall hydrolase of alkalophilic Bacillus subtilis BL-29 was cloned into E. coli JM109 using pUC18 as a vector. A recombinant plasmid, designated pCWL45B, was contained in the fragment originating from the alkalophilic B. subtilis BL-29 chromosomal DNA by Southern hybridization analysis. The nucleotide sequence of a 1.6-kb HindIII fragment containing a cell wall hydrolase-encoding gene was determined. The nucleotide sequence revealed an open reading frame (ORF) of 900 bp with a concensus ribosome-binding site located 6 nucleotide upstream from the ATG start codon. The primary amino acid sequence deduced from the nucleotide sequence revealed a putative protein of 299 amino acid residues with an M.W. of 33, 206. Based on comparison of the amino acid sequence of the ORF with amino acid sequences in the GenBank data, it showed significant homology to the sequence of cell wall amidase of the PBSX bacteriophage of B. subtilis.

• Applied Biological Chemistry
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• v.50 no.2
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• pp.87-94
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• 2007

A bacterium which has high enzymatic activities such as amylase, cellulase and protease was isolated from Korean traditional soybean food, doenjang. The isolated bacterium was identified to Bacillus subtilis HS25 by the test of morphological and biochemical properties according to Bergey's Manual of Systematic Bacteriology and API 50 CHL kit, and by the 16S rDNA sequence. The isolated B. subtilis HS25 had a potent antibacterial activity against food born causative or pathogenic bacteria. B. subtilis HS25 is endospore forming cell and contained flagella and abundant viscous material at the out layer of cell wall. It was rod type bacterium $(0.5{\sim}0.8{\times}3{\sim}5{\mu}m)$ having biochemical characteristics such as gram staining(+), catalase(+), oxidase(-) and hydrolysis of esculin(+). The optimal medium compositions for production of antibacterial substance in the B. subtilis HS25 were 1% of soluble starch, 0.5% of yeast extract, 0.5% of peptone and 0.05% of MgCl$_2{\cdot}6H_{2}O$. The optimum temperature and pH of the growth of the B. subtilis HS25 was 35$^{\circ}C$ and pH 7.5, respectively. The antibacterial activity was more high in neutral to a little alkaline pH (6.5-10.5) than in acidic pH. The optimal shaking speed to grow and to produce antibacterial substance of the B. subtilis HS25 was 160${\sim}$200 rpm. The optimal culture time for antibacterial activities of the bacterium were shown to be in the range of 12-36 hr.

• BMB Reports
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• v.45 no.4
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• pp.239-241
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• 2012

Iron availability is limited in the environment and most bacteria have developed a system to acquire iron from host hemoproteins. Heme oxygenase plays an important role by degrading heme group and releasing the essential nutrient iron. The structure of Bacillus subtilis HmoB was determined to 2.0 ${\AA}$ resolution. B. subtilis HmoB contains a typical antibiotic biosynthesis monooxygenase (ABM) domain that spans from 71 to 146 residues and belongs to the IsdG family heme oxygenases. Comparison of HmoB and IsdG family proteins showed that the C-terminal region of HmoB has similar sequence and structure to IsdG family proteins and contains conserved critical residues for heme degradation. However, HmoB is distinct from other IsdG family proteins in that HmoB is about 60 amino acids longer in the N-terminus and does not form a dimer whereas previously studied IsdG family heme oxygenases form functional homodimers. Interestingly, the structure of monomeric HmoB resembles the dimeric structure of IsdG family proteins. Hence, B. subtilis HmoB is a heme oxygenase with a novel structural feature.

• Korean Journal of Microbiology
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• v.42 no.4
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• pp.313-318
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• 2006

A thermophilic bacterium producing the extracellular cellulase was isolated from soybean paste, and the isolate WL-12 has been identified as Bacillus licheniformis on the basis on its 16S rRNA sequence, morphology and biochemical properties. A gene encoding the cellulase of B. licheniformis WL-12 was cloned and its nucleotide sequence was determined. This cellulase gene, designated celA, consisted of 1,551 nucleotides, encoding a polypeptide of 517 amino acid residues. The gene product contained catalytic domain and cellulose binding domain. The deduced amino acid sequence was highly homologous to those of cellulases of B. licheniformis, B. subtilis and B. amytoliquefaciens belonging to the glycosyl hydrolase family 5. When the celA gene was highly expressed using a strong B. subtilis promoter, the extracellular cellulase was produced up to 7.0 units/ml in B. subtilis WB700.

• Korean Journal of Microbiology
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• v.49 no.3
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• pp.262-269
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• 2013

The object of this study was to improve the quality of Cheonggukjang with new starter, Bacillus subtilis BC-P1. Twenty strains were isolated from the commercial cheonggukjang and 1 Bacillus strain (BC-P1) with protease activity was selected. The 16S rRNA gene sequence revealed that the BC-P1 was closely related to B. subtilis with 99% homology. The quality characteristics of chunggukjang fermented with B. subtilis BC-P1, Bacillus nato (PC) and commercial chunggukjang (NC) were investigated. The characteristics of fermentation were determined by protease, lipase, xylanase, chitinase, and fibrinolytic activities, reducing sugar, nutrient composition and amino acid contents of cheonggukjang sample. Cheonggukjang fermented with B. subtilis BC-P1 showed the strongest fibrinolytic, xylanase, and chitinase activities. Reducung sugar contents of Cheonggukjang samples were $30.16{\pm}2.11$ mg/g (NC), $28.56{\pm}1.52$ mg/g (PC), $32.39{\pm}1.87$ mg/g (BC-P1). And their total amino acid contents were 338.99 mg% (NC), 445.19 mg% (PC), 741.35 mg% (BC-P1). These results suggested that B. subtilis BC-P1 was suitable to be used as a starter to enhance the quality and effects of cheonggukjang.

• Korean Journal of Food Science and Technology
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• v.38 no.4
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• pp.548-553
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• 2006

The aim of this study was to produce Chungkookjang-a food produced through fermentation with Bacillus licheniformis E1-that contains an increased concentration of a bacterial biological response modifier (B-BRM). Unfortunately, sensory studies have indicated that B. licheniformis E1-fermented Chungkookjang is unacceptable for commercial use. We isolated another bacterial strain from this food product: B. subtilis S2. The optimum time and temperature for Chungkookjang fermentation with B. licheniformis E1 and B. subtilis S2 were 48 hr and $40^{\circ}C$, respectively. Sensory studies showed that Chungkookjang fermented by both B. licheniformis E1 and B. subtilis S2 was more acceptable than B. licheniformis E1 only. The amino nitrogen and crude protein content of the product were 359 mg% and 45.6% respectively. Additionally, it was confirmed that the proliferation of mouse splenic lymphocytes increased significantly, when the cells were treated with the BRM from Chungkookjang fermented using the mixture of bacterial strains in vitro. These results suggest that the enriched Chungkookjang may help patients who are medically in need of potentiation of lymphocytes proliferation.

• Korean Journal of Food Science and Technology
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• v.44 no.3
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• pp.312-316
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• 2012

In order to understand the effect of hydrostatic pressure (HP) on Bacillus subtilis isolated from makgeolli, the survival of B. subtilis after HP treatment (400 MPa for 5 min) in various substrates including phosphate buffer, tryptone soya broth at pH 7 and 4, and makgeolli at pH 4 was evaluated depending on bacterial forms (spores and vegetative cells) and adaptation conditions ($25^{\circ}C$ for 3 h, or $10^{\circ}C$ for 24 h). Spores were generally resistant to HP (<1 log reduction) regardless of conditions. In contrast, vegetative cells were generally susceptible to HP (up to 3 log reduction-except makgeolli) and were more susceptible after 3 h at $25^{\circ}C$ compared to 24 h at $10^{\circ}C$. In vegetative cells inoculated makgeolli (7 log CFU/mL), the colonies were not detected after 24 h at $10^{\circ}C$. Consequently, B. subtilis in makgeolli easily existed as spores and the spores were resistant to HP. Results demonstrate that HP was more promising in the inactivation of vegetative cells.

• Journal of Life Science
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• v.23 no.4
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• pp.560-568
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• 2013

Chungkookjang was fermented by B. subtilis MC31, a ${\gamma}$-amino butyric acid (GABA) producing microorganism. The characteristics of Chungkookjang were investigated while fermenting. Twenty four amino acids were detected in Chungkookjang, leucine was the highest of them all. Total cell populations of B. subtilis MC31 phase were between log $9.52{\pm}0.5$ ~ log $9.049{\pm}0.5$ CFU/g at stationary phase. Contents of moisture, crude ash, crude protein, crude lipid and crude fiber are $61.07{\pm}0.01%$, $1.52{\pm}0.01%$, $17.66{\pm}0.04%$, $8.96{\pm}0.03%$ and 2.61%, respectively. Contents of ammonia type nitrogen, amino type nitrogen and reducing sugar were increased during fermentation at $40^{\circ}C$ for 72 hr, however those of titratable acidity and total sugar were decreased. pH was slowly alkalized during fermentation. Viscous substance and protease contents in Chungkookjang were $4.7{\pm}0.05%$ and $0.519{\pm}7.36$ g/l, apiece. When the fibrin plate and Robbin method for fibrinolytic activity were applied, B. subtilis MC31 showed high activity. These results suggested that B. subtilis MC31 is suitable to be used as a starter to enhance the quality of Chungkookjang.

• Journal of fish pathology
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• v.10 no.2
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• pp.125-135
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• 1997

The possibility of identification of families of antibacterial agent residues in fish tissue was studied by disk assay using three test organisms, Bacillus subtilis BGA, Micrococcus luteus ATCC 9341, and Bacillus cereus var. mycoides ATCC 11778. In the present method, a simple clean-up procedure was performed to obtain the aqueous solution from homogenized flounder muscle sample(10g) in Mcilvaine buffer. Then, aqueous solution was fractionated into A and B to be used in disk assay by choloroform and Sep-Pak $C_{18}$ cartridge column after being defatted in hexane. The chloroform layer of fraction A was used for the analysis of macrolide antibiotics(ML), sulfa drugs(SA), chloramphenicol(CP), and quinolone antibiotics(QN). Adsorbed materials to Sep-Pak $C_{18}$ of fraction B were also employed for the analysis of penicillins(PC), tetracyclines(TC), and nitrofuran derivatives(NF) Minimun-detectable concentrations by the present method were, $0.1{\mu}g$/g for oxytetracycline, tetracycline, doxycycline, spiramycin and ciprofloxacin, $0.025{\mu}g$/g for erythromycin and ampicillin, $1.0{\mu}g$/g for sodium nifurstyrenate and florfenical, $0.25{\mu}g$/g for sulfamonomethoxie and sulfadimethoxine, $2.5{\mu}g$/g for oxolinic acid and flumequine, and $15{\mu}g$/g for piromidic acid, respectively. Three test organisms showed different sensitivity patterns for each family of antibacterial agent. Sensitivity patterns were B. cereus > B. subtilis > M. luteus for TC and NF, M. luteus > B, subtilis > B. cereus for ML and PC, B. cereus = B. subtilis > M. luteus for CP and QN, and B. subtilis > B. cereus＝M. luteus for SA. The present method utilizing these characteristics could be useful as a routine screening test for the determination of family of antibacterial agent residues in fish tissue.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.5
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• pp.846-851
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• 1998

Characteristics of chunggugjang fermented by Bacillus subtilis DC-2, a pigment producing bacterium, were investigated. More water soluble browning materials were produced with fermentation time. The pH was gradually alkalized. The contents of amino nitrogen were extraordinarily increased with fermentation time. Both strength and hardness were gradually decreased during fermentation. Total 30 volatile compounds were identified in the chunggugjang fermented by B. subtilis DC-2. The pyrazines were detected more than any other compounds. The good aroma of the chunggujang fermented by B. subtilis DC-2 was considered to be contributed by tetramethylpyrazine, trimethylpyrazine, 1-octen-3-ol, 2, 5-dimethylpyrazine and guaiacol.