• Title, Summary, Keyword: Bacillus subtilis

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Isolation and Characterization of Bacillus subtilis CA105 from Spent Mushroom (Pleurotus ostreatus) Substrates (느타리버섯 수확후배지로부터 분리한 Bacillus subtilis CA105의 특성)

  • Kim, Hye Soo;Kim, Chul Hwan;Kwon, Hyun Sook;Lee, Chan-Jung;Kong, Won-Sik;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.4
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    • pp.305-309
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    • 2015
  • In order to isolate compost-promoting bacteria with high activity of cellulase and xylanase, spent mushroom substrates with sawdust were collected from mushroom cultivation farm, Jinju, Gyeongnam in Korea. Among of the isolates, one strain, designated CA105 was selected by agar diffusion method. The strain CA105 was identified as members of the Bacillus subtilis by biochemical characteristics using VITEK 2 system. Comparative 16S rRNA gene sequence analysis showed that isolate CA105 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rRNA gene sequence similarity of 98.9%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, isolate CA105 was classified within the genus Bacillus subtilis, for which the name Bacillus subtilis CA105 is proposed. The cellulase and xylanase activity of B. subtilis CA105 was slightly increased according to bacterial population from exponential phase to stationary phase in growth curve for Bacillus sp. CA105.

Purification and Characterization of an Antimicrobial Substance from Bacillus subtilis HH28 Antagonistic to Bacillus cereus (Bacillus cereus를 억제하는 Bacillus subtilis HH28의 항균물질 정제와 특성규명)

  • Cha, Hyun A;Chung, Dawn;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.393-401
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    • 2014
  • A bacterium producing antimicrobial substance was isolated from cheonggukjang. The bacterium was identified as a strain of Bacillus subtilis by 16S rDNA sequencing and designated as Bacillus subtilis HH28. The antimicrobial substance produced from Bacillus subtilis HH28 was purified by 0-80% ammonium sulfate precipitation, DEAE-sepharose FF column chromatography, and Sephacryl S-200 HR gel chromatography. The molecular weight of the purified antimicrobial substance was estimated to be approximately 3,500 Da using Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis and direct detection analysis. Antimicrobial substance from B. subtilis HH28 not only inhibited B. cereus, but also Listeria monocytogenes and Vibrio parahaemolyticus. The purified antimicrobial substance was stable at $40-80^{\circ}C$, and between pH 2 and 8. Antimicrobial activity of the purified substance was completely destroyed by treatment of protease, proteinase K, and pronase E, indicating that it is proteinaceous.

Identification of a Newly Isolated Protease-producing Bacterium, Bacillus subtilis FBL-1, from Soil (토양으로부터 새로이 분리된 단백질 분해효소 생산 미생물 Bacillus subtilis FBL-1의 동정)

  • Kim, Mina;Si, Jin-Beom;Wee, Young-Jung
    • 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.

Quality characteristics of popped rice Doenjang prepared with Bacillus subtilis strains (Bacillus subtilis 균주를 이용하여 제조한 팽화미 된장의 품질 특성)

  • Lee, Kyung Ha;Kim, Eun Ju;Choi, Hye Sun;Park, Shin Young;Kim, Jae Hyun;Song, Jin
    • Korean Journal of Food Preservation
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    • v.22 no.4
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    • pp.545-552
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    • 2015
  • This study investigated the quality characteristics of popped rice Doenjang prepared with different Bacillus strains (Bacillus subtilis KACC 15935, and Bacillus subtilis HJ18-9). The changes in the enzyme activity (protease, cellulase, and ${\alpha}$-amylase), amino-type nitrogen and ammonia-type nitrogen contents, and the reducing sugar were investigated during the fermentation period. Enzymes such as protease, cellulase, and a-amylase plays an important role in the changes in composition of nutrients, and in flavor and taste of popped rice Doenjang. Protease activities of the popped rice deonjang fermented with different Bacillus strains (control, B. subtilis KACC 15935, and B. subtilis HJ18-9) was in the range of 171.77-185.97 unit/g at the beginning of fermentation, and there were no significant differences among the samples. On the other hand, the protease activity in popped rice Doenjang fermented with B. subtilis HJ18-9 increased significantly up to $248.77{\pm}4.53unit/g$ at the end of fermentation (p<0.05). Cellulase activity and a-amylase activity of popped rice Doenjang in HJ18-9 was higher than these of other samples. After 56 days of fermentation, amino-type nitrogen in popped rice deonjang fermented with control, B. subtilis KACC 15935, and B. subtilis HJ18-9 increased significantly up to $174.99{\pm}3.70$, $166.59{\pm}1.40$, $225.39{\pm}3.70mg%$, respectively (p<0.05). These results suggested that B. subtilis HJ18-9 was a suitable starter for the preparation of soybean paste.

Antimicrobial Activity of Water Extract of Green Tea against Cooked Rice Putrefactive Microorganism (쌀밥 부패미생물에 대한 녹차 물추출물의 항균 활성)

  • Roh, Hyun-Jeong;Shin, Yong-Seo;Lee, Kap-Sang;Shin, Mee-Kyung
    • Korean Journal of Food Science and Technology
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    • v.28 no.1
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    • pp.66-71
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    • 1996
  • To extend shelf life of cooked rice, main putrefactive microorganism isolated from cooked rice were identified by using the API 50 CHB kit and fatty acid analysis of the cell and antimicrobial activity of water extract of green tea was tested against isolated strains and some type of strains. The growths of Bacillus subtilis ATCC 6633, Salmonella typhimurium ATCC 14028, Bacillus cereus YUFE 2004 and Staphylococcus aureus YUFE 2087 were inhibited in broth containing 500 and 1000 ppm of green tea extract. Main putrefactive microorganisms of cooked rice were identified as Bacillus subtilis RHJ-I and Bacillus subtilis RHJ-II. Green tea extract of 500 and 1000 ppm level inhibited the growth of Bacillus subtilis RHJ-I only.

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Characterization of the Functional Properties of Soy Milk Cake Fermented by Bacillus sp.

  • Oh, Soo-Myung;Kim, Chan-Shick;Lee, Sam-Pin
    • Food Science and Biotechnology
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    • v.15 no.5
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    • pp.704-709
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    • 2006
  • The mucilage production and tyrosine content in soy milk cake (SMC) fermented by Bacillus firmus NA-1, Bacillus subtilis GT-D, and B. subtilis KU-A was improved by fortification with 10% defatted soybean flour. The fibrinolytic activity and consistency of the SMC were drastically increased by solid-state fermentation for 1 day. However, the consistency of the fermented SMC gradually decreased during fermentation for 3 days. Furthermore, the tyrosine content of the freeze-dried powder of SMC fermented by three Bacillus sp. was 9 times higher than that of unfermented SMC. The soybean proteins, including the 7S and 11S subunits, were partially digested during alkaline fermentation, producing lower molecular-weight peptides. The fibrinolytic enzyme produced in SMC fermented by B. firmus NA-l and B. subtilis KU-A exhibited higher thermal stability than that of B. subtilis GT-D fermentation. The powder obtained from B. subtilis GT-D fermentation had an ${\alpha}$-amylase activity and lower consistency compared to those of B. firmus NA-1 and B. subtilis KU-A. In addition, this powder contained 6.3% moisture content, 27% crude protein content and 9 units of fibrinolytic activity and proteolytic activity.

Separation and Purification of Protease from Bacillus subtlils CCKS-111 in Korean Traditional Soy Sauce (한국재래간장으로 부터 분리한 Bacillus subtilis CCKS-111이 생성하는 Protease의 분리 및 정제)

  • Kim, Sung;Lim, Seong-Il;Lee, Hee-Duck;Lee, Seon-Ho;Son, Jun-Ho;Choi, Hee-Jin;Kim, Yeung-Hweal;Choi, Cheong
    • Applied Biological Chemistry
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    • v.40 no.3
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    • pp.178-183
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    • 1997
  • A protease was purified from Bacillus subtilis CCKS-111 by ammonium sulfate treatment, DEAE-cellulose ion-exchange chromatography, Sephadex G-100 gel filtration and high performance liquid chromatography (HPLC). The specific activity of the purified enzyme was 24.3 unit/mg protein and the purification fold of enzyme was 50.6. Molecular weight of the purified enzyme estimated about 28,000 by HPLC gel filtration. The amino acid residues of this enzyme were 251.3 except threonine, serine and glycine. This result was similar to Bacillus subtilis subtilisin DY. From the first N-terminal amino acid to the 32th amino acid, the amino acid sequence was estimated after RP-HPLC elution. N-terminal and the 32th amino acids were alanine and aspartic acid. Alanine, serine, glycine and arginine were four major acids in the enzyme.

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Variation of fibrinolytic enzyme activity produced Bacillus subtilis by gene cloning (유전자 cloning에 의한 Bacillus subtilis의 fibrinolytic enzyme 활성 변화)

  • 이홍석;유천권;이철수;강상모
    • Microbiology and Biotechnology Letters
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    • v.28 no.1
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    • pp.14-20
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    • 2000
  • The transformation of Bacillus subtilis K-54 and J-10 was carried out with constructed vectors containing structure and enhancer genes of aprN and prtR, to increase their fibrinolytic enzyme activity. Bands for the aprN and prtR genes were identified from B. subtilis J-10 by PCR that was carried out with the constructed primers for the genes. In addition, the gene fragments contained promoter site based on the results of analysing their nucleotide sequence. The two gene fragments, aprN and prtR, obtained by the PCR, were, then, inserted to vector such as T-vector and E.coli/Bacillus shuttle vector. The constructed vector were designated as pAPR2 (aprN), pENC2 (prtR) and pFLA1 (aprN and prtR), respectively. The constructed vector was used for transformation of the strains of B.subtilis J-10 and B. subtilis K-54 and the fribrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and the fibrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and pFLA1, resulted in the increase of fibrinolyitic enzyme activity in B. subtilis J-10 by 27.3% and 16%, respectively. However, the introduction of pENC2 to B. subtilis J-10 did not seem to induce increase of the enzyme activity. The strain of B.subtilis K-54 transformed with pENC2 showed an increased fibrinolytic activity by 5 folds compared with that of the original strain of B. subtilis K-54.

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Controlling Activity of Bacillus subtilis KB-401 against Cucumber Powdery Mildew Caused by Sphaerotheca fusca (오이 흰가루병에 대한 Bacillus subtilis KB-401의 방제 효과)

  • Nam, Myung-Hyeun;Choi, Jae-Pil;Kim, Hyung-Jo;Lee, Jae-Jun;Lim, Keun-Hwan;Kim, Young-Gwon;Kim, Heung-Tae;Jeun, Yong-Chull
    • The Korean Journal of Pesticide Science
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    • v.14 no.1
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    • pp.49-53
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    • 2010
  • Disease control efficacy was evaluated with use of Bacillus subtilis KB-401 against cucumber powdery mildew in a greenhouse and fields. B. subtilis KB-401 showing inhibitory effect on mycelial growth of various phytopathogenic fungi was formulated for the evaluation. The formulated biofungicide of B. subtilis KB-401 was less effective at 1,000 times dilution rate than that at 250 or 500 times dilution rate. The powdery mildew was successfully controlled by the biofungicide at the early stage of disease development. The field performance of the biofungicde was conducted in Asan and Cheonan city. Three or four consecutive applications of the biofungicide at 500 dilution rate with 10-day intervals resulted in considerable efficacy of disease control as high as 83.3%.

Antiobesity Effect of the Bacillus subtilis KC-3 Fermented Soymilk in 3T3-L1 Adipocytes (3T3-L1 지방세포에서 Bacillus subtilis KC-3 발효두유의 항비만 효과)

  • Kim, Ji-Young;Jeong, Jung-Eun;Moon, Suk-Hee;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1126-1131
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    • 2010
  • The antiobesity effect of soymilks fermented with Bacillus subtilis KC-3 (KCCM 42923) from cheonggukjang was compared with other sources of B. subtilis KCCM 11316 and B. subtilis MYCO. The antiobesity effect was investigated by measuring the release of leptin, Oil red O staining, glycerol secretions and adipogenic transcription factor by reverse transcription-polymerase chain reaction (RT-PCR) in the 3T3-L1 adipocytes. Fermented soymilk with B. subtilis KC-3 (F-KC) led to decrease levels of leptin secretion and increase levels of glycerol secretion in the cells. In addition, F-KC reduced contents of Oil red O dye in the 3T3-L1 adipocytes. Also, mRNA expression levels of both SREBP-1c (sterol regulatory element-binding protein 1-c) and PPAR-$\gamma$ (peroxisome proliferator-activated receptor-$\gamma$), which are adipogenic transcription factor, in cells treated with F-KC were markedly down regulated. These results demonstrate that the Bacillus subtillis fermented soymilk (F-KC) decreased lipid content in 3T3-L1 adipocytes by inhibiting lipogenesis. All B. subtilis fermented soymilks had shown antiobesity activities, however, F-KC exhibited the strongest antiobesity effect in the 3T3-L1 adipocytes. Our study suggests that especially F-KC increased the potential of antiobesity effects.