• Title/Summary/Keyword: Bacillus subtilis

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Cloning and Characterization of a Cellulase Gene from a Plant Growth Promoting Rhizobacterium, Bacillus subtilis AH18 against Phytophthora Blight Disease in Red-Pepper (고추역병을 방제하는 PGPR균주 Bacillus subtilis AH18의 항진균성 Cellulase 유전자의 Cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Jung, Hee-Kyoung;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.311-317
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    • 2006
  • Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

Isolation of Biogenic Amines-Degrading Strains of Bacillus subtilis and Bacillus amyloliquefaciens from Traditionally Fermented Soybean Products (전통장류에서 Biogenic Amines 분해 능력을 가지는 Bacillus subtilis 및 Bacillus amyloliquefaciens 균주의 분리)

  • Kim, Yong-Sang;Cho, Sung-Ho;Jeong, Do-Yeon;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.48 no.3
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    • pp.220-224
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    • 2012
  • In order to reduce harmful biogenic amines in the traditionally fermented soybean products, five isolates with biogenic amines-degrading property were obtained from 83 traditionally fermented soybean products. The strains were found to reduce biogenic amines including histamine, tyramine, putrescine, and cadaverine by 27 to 92% in the cooked soybean containing 5.3% of each biogenic amine over 10 days of incubation. The morphological and biochemical tests and the phylogenetic relationships based on 16S rRNA gene sequences indicated that the five isolates were most closely related to Bacillus subtilis or B. amyloliquefaciens. The use of selected strains would be a potential control measure in manufacturing traditionally fermented soybean products that are difficult to control biogenic amine levels.

Cloning and Expression of A Bacillus licheniformis Cellulase Gene (Bacillus licheniformis WL-12의 cellulase 유전자 클로닝과 발현)

  • Yoon, Ki-Hong
    • 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.

Isolation of Bacillus sp. Producing Multi-enzyme and Optimization of Medium Conditions for Its Production Using Feedstuffs for Probiotics (Probiotics용 복합효소 분비 Bacillus sp.의 분리 및 원료사료를 이용한 균주 생산을 위한 배지 조건의 최적화)

  • 양시용;송민동;김언현;김창원
    • Microbiology and Biotechnology Letters
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    • v.29 no.2
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    • pp.110-114
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    • 2001
  • Isolation of BacilLus sp. producing multi-enzyme and optimization of medium conditions for its production using feedstuffs for probiotics were carried out in this study. A bacterium isolated from natural resources, namely Bacillus subtilis 4-3, has multi-enzyme activity (phytase. cellulase, xylanasc, protease, and amylase. In the culture of B. subtilis 4-3 using soybean meal and rice bran. relatively low phytate degradation was noted using whereas high phytate degradability was observed with wheat bran (80.63%). The optimal composition of medium using feedstuffs was 1.0% (w/v) soybean meal and 2% (w/v) molasses to yield high cell growth.

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Synthesis and Antimicrobial Activity of Polyacryloyl- and Polymethacryloylcephradine (Polyacryloyl- 및 Polymethacryloylcephradine의 합성과 항균작용에 관한 연구)

  • Kim, Sun-Il;Cha, Wol-Suk;Na, Jae-Woon;Kim, Young-Ho;Ko, Ok-Hyun
    • Journal of the Korean Chemical Society
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    • v.36 no.2
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    • pp.282-286
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    • 1992
  • The reaction of N-hydroxysuccinimide with polyacrylic acid and polymethacrylic acid gave poly(N-acryloxysuccinimide)and poly(N-methacryloxysuccinimide), whose reaction with cephradine provided polyacryloylcephradine and polymethacryloylcephradine. The activities of these polymeric drugs were investigated in terms of minimum inhibitory concentration by the common twofold dilution technique. Polyacryloylcephradine revealed excellent antibacterial activity against Staphylococcus aureus ATCC 25923, Staphylococcus aureus FDA 209P, Bacillus subtilis ATCC 6633, Bacillus licheniformis ATCC 14580, Escherishia coli BE 1186 and Salmonella typhimurium TV 119. Polymethacryloylcephradine revealed excellent Staphylococcus aureus ATCC 25923, Staphylococcus aureus FDA 209P, Bacillus subtilis ATCC 6633, Escherichia coli Be 1186 and Salmonella typhimurium TV 119.

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Solubility, Viscosity, Water Holding Capacity, and Oil Holding Capacity of Soybean Proteins by Bacillus subtilis and/or Lactobacillus bulgaricus (Bacillus subtilis와 Lactobacillus bulgaricus에 의한 청국장 단백질의 용해성, 점성, 보수성 및 보유성)

  • Lee, Jin-Woo
    • 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.

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Volatile Components of Korean Soybean Paste Produced by Bacillus subtilis PM3

  • JONG-KYU KIM;JI, WON-DAE;SUNG-HO YANG;MYEONG-RAK CHOI
    • Journal of Microbiology and Biotechnology
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    • v.5 no.3
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    • pp.143-148
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    • 1995
  • A strain producing soybean paste flavor was isolated from traditional Korean soybean paste. The isolate was identified as Bacillus subtilis PM3. The neutral fraction representing the traditional soybean paste aroma was obtained from the whole volatile components produced by B. subtilis PM3 in cooked soybean. Each separated peak from the neutral fraction of gas chromatogram was identified by gas chromatography-mass spectrometry (GC/MS) and Kovat's retention index, and the aromas of each peak were investigated by a sniffing test with the exercise panel. The twenty-nine components, including six character impact compounds and twelve components of flavors of Korean soybean paste, were confirmed. Some regions of gas chromatogram represented the soybean paste odor. It has been confirmed that traditional Korean soybean paste can be manufactured with the strain B. subtilis PM3.

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Molecular Cloning and Sequencing of Cell Wall Hydrolase Gene of an Alkalophilic Bacillus subtilis BL-29

  • Kim, Tae-Ho;Hong, Soon-Duck
    • 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.

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Conserved Virulence Factors of Pseudomonas aeruginosa are Required for Killing Bacillus subtilis

  • Park Shin-Young;Heo Yun-Jeong;Choi Young-Seok;Deziel Eric;Cho You-Hee
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.443-450
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    • 2005
  • The multi-host pathogen, Pseudomonas aeruginosa, possesses an extraordinary versatility which makes it capable of surviving the adverse conditions provided by environmental, host, and, presumably, competing microbial factors in its natural habitats. Here, we investigated the P. aeruginosa-Bacillus subtilis interaction in laboratory conditions and found that some P. aeruginosa strains can outcompete B. subtilis in mixed planktonic cultures. This is accompanied by the loss of B. subtilis viability. The bactericidal activity of P. aeruginosa is measured on B. subtilis plate cultures. The bactericidal activity is attenuated in pqsA, mvfR, lasR, pilB, gacA, dsbA, rpoS, and phnAB mutants. These results suggest that P. aeruginosa utilizes a subset of conserved virulence pathways in order to survive the conditions provided by its bacterial neighbors.