• Title/Summary/Keyword: subtilis

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Construction of Pretense-defective Mutant of Bacillus subtilis by Homologous DNA Recombination (상동성 유전자재조합을 이용한 단백질분해효소 비생산 바실러스균주의 구축)

  • Lee, Jin-Tae;An, Bong-Jeun
    • Food Science and Preservation
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    • v.7 no.4
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    • pp.414-417
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    • 2000
  • Competent cell transformation of B. subtilis AC819 was carried out using phenotypic protease-defective(Npr-) DNA of B. subtilis MT-2. An obtained transformant, designated B. subtilis HL-1, was obtained by homologous DNA recombination. Phenotypes of B. subtilis HL-1 were characterized histidine requirement streptomycin-resistance, tetracyclin resistance and non-producing protease. Protoplast transformation frequency of B. subtilis HL-1 by plasmid pUB110 was higher than that of B. subtilis MT-2. From this result, B. subtilis HL-1 is useful for protease gene transformation and thermostable protease gene cloning as a host.

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Enhancement of Bacteriocin Production by Bacillus subtilis cx1 in the Presence of Bacillus subtilis ATCC6633 (Bacillus subtilis ATCC6633이 Bacillus subtilis cx1의 박테리오신 생산에 미치는 유도효과)

  • Chang Mi;Chang Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.221-227
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    • 2006
  • BSCX1 was an antimicrobial peptide produced by Bacillus subtilis cx1. Attempts were made to determine the location of inducing factor in the bacteriocin-sensitive cell affecting bacteriocin BSCX1 production. Mixed culture of the bacteriocin producer strain B. subtilis cx1 and its sensitive strain B. subtilis ATCC6633, increased production of bacteriocin BSCX1. The result suggested the presence of a bacteriocin inducing factor in the sensitive strain. The inducing factor was localized in the cell debris and intracellular fraction of B. subtilis ATCC6633. Bacteriocin BSCX1 inducing factor was found to be highly stable in the pH range 2.5-9.5, but inactivated within 3h over $50^{\circ}C$, and treatment with proteinase K destroyed its inducing activity, this result suggested that the inducing factor should be a proteinaceous nature.

Cloning and Strong Expression of a Bacillus subtilis WL-3 Mannanase Gene in B. subtilis

  • Yoon, Ki-Hong;Lim, Byung-Lak
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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    • pp.1688-1694
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    • 2007
  • A gene encoding the mannanase of Bacillus subtilis WL-3, which had been isolated from Korean soybean paste, was cloned into Escherichia coli and the nucleotide sequence of a 2.7-kb DNA fragment containing the mannanase gene was subsequently determined. The mannanase gene, designated manA, consisted of 1,080 nucleotides encoding a polypeptide of 360 amino acid residues. The deduced amino acid sequence was highly homologous to those of mannanases belonging to glycosyl hydrolase family 26. The manA gene was strongly expressed in B. subtilis 168 by cloning the gene downstream of a strong B. subtilis promoter of plasmid $pJ27{\Delta}88U$. In flask cultures, the production of mannanase by recombinant B. subtilis 168 reached maximum levels of 300 units/ml and 450 units/ml in LB medium and LB medium containing 0.3% locust bean gum, respectively. Based on the zymogram ofthe mannanase, it was found that the mannanase produced by recombinant B. subtilis could be maintained stably without proteolytic degradation during the culture time.

Isolation and Identification of Antimicrobial Agent Producing Microorganisms and Sensitive Strain from Soil (토양으로부터 항균물질 생성균 및 감수성 균주의 분리 및 동정)

  • 장해춘;김수인;김인철
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.3
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    • pp.526-533
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    • 1999
  • Two species of antimicrobial agent producing bacteria and one sensitive strain were isolated from soil. Those were identified as B. subtilis, B. licheniformis, and Curtobacterium sp. by morphological, biochemical, physiological and chemotaxonomic characteristics. These were designated as B. subtilis cx1, B. licheniformis cy2 and Curtobacterium sp. cf3, respectively. Antimicrobial agent produced by B. subtilis cx1 showed high antibacterial activity against gram positive bacteria including of B. subtilis, Curtobacterium sp., L. mesenteroids, Staphy. aureus, S. faecalis and even gram negative bacteria, P. aeruginosa. Antimicrobial agent from B. licheniformis cy2 showed slightly lower antimi crobial activity than that from B. subtilis cx1. These two strains showed maximum production of antimicrobial agents at 30oC for 9~21hr cultivation. Curtobacterium sp. cf3 showed more sensitive activity than a sensitive strain of B. subtilis ATCC 6633 which was same genus or species with the B. subtilis cx1 and B. subtilis cy2, when the antimicrobial agent producing strains, B. subtilis cx1 and B. subtilis cy2, were directly applied onto these sensitive lawns.

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Selection of Beneficial Microbial Agents for Control of Fungal Diseases in the Phyllosphere of Cucumber Plant (오이 지상부의 주요 곰팡이 병해의 생물적 방제용 유용미생물의 선발)

  • Lee, Sang-Yeob;Lee, Young-Kee;Park, Kyung-Seok;Kim, Yong-Ki
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.326-331
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    • 2010
  • Bacillus subtilis B29, B. subtilis M10 and Streptomyces sp. CC19 obtained from phyllosphere of cucumber plants were selected for biological control of fungal air-borne diseases. For the downy mildew, diseased area of B. subtilis B29, B. subtilis M10 and Streptomyces sp. CC19 showed 0.5%, 20.2% and 42.0%, but that of control was 82.0% respectively, in cucumber seedling test. Incidence of powdery mildew by once application of B. subtilis B29, B. subtilis M10 and Streptomyces sp. CC19 was 2.8%, 3.6% and 12.3%, respectively, whereas that of control was 65.6%. On the gray mold, diseased area of B. subtilis B29, B. subtilis M10 and Streptomyces sp. CC19 was 8.0%, 30.8% and 5.2%, respectively, compared to 81.2% for the control. Therefore, B. subtilis B29 could be a prospective antagonist for biological control of powdery mildew, downy mildew and gray mold of cucumber plant.

Comparison of Characteristics of Koji Manufactured with Bacillus subtilis B-4 and Aspergillus oryzae F-5 (Bacillus subtilis B-4와 Aspergillus oryzae F-5로 제조한 코오지의 특성 비교)

  • Kwon, Dong-Jin
    • Korean Journal of Food Science and Technology
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    • v.34 no.5
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    • pp.873-878
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    • 2002
  • In order to industrialize traditional Meju-tasting Koji, the characteristics of Koji manufactured with bacteria and fungi found in traditional Meju were investigated. Bacillus subtilis B-4 and Aspergillus oryzae F-5 showing high enzyme activities including those of amylase and protease were used. L-value of Koji manufactured with B. subtilis B-4 had darker color and higher enzyme production than A. oryzae F-5 made one. B. subtilis B-4 made Koji showed higher enzyme production and sensony evaluation score than A. oryzae F-5 Koji. A. oryzae F-5 Koji showed superior color to B. subtilis B-4 Koji. Activity in color, capacity of enzyme production, viable cell count, and sensory evaluation of water activity controlled Koji was superior to the uncontrolled one.

Mixture of Edwardsiella tarda specific Bacteriophage and Bacillus subtilis KM-1enhanced bactericidal activity against Edwardsiella tarda (Edwardsiella tarda의 특이 Bacteriophage와 Bacillus subtilis KM-1혼합액이 Edwardsiella tarda 에 미치는 항균효과)

  • Baek, Min Suk;Hwang, Yo Sep;Choi, Sanghoon
    • Journal of fish pathology
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    • v.26 no.3
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    • pp.185-191
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    • 2013
  • The present study was performed to investigate an antibacterial activity of specific bacteriophage (phage) and Bacillus subtilis KM-1 (B. subtilis) mixture against Edwardsiella tarda (E. tarda). An appropriate number of phage showing the most effective antibacterial activity was $2{\times}10^5$ PFU/ml with $1{\times}10^7$ CFU/ml of B. subtilis 36 h post incubation. On the other hand, B. subtilis showed a dose dependant manner in inducing antibacterial activity in the presence of phage ($2{\times}10^5$ PFU/ml). The phage and B. subtilis mixture showed higher antibacterial activity against E. tarda than phage or B. subtilis only. These results suggest that the phage and B. subtilis mixture could be utilized as an alternative to antibiotics in the control of fish diseases caused by E. tarda.

Characteristics of Cadmium-Resistant Bacillus subtilis DT134 (Bacillus subtilis DT134의 카드뮴 저항성)

  • 윤경표
    • KSBB Journal
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    • v.13 no.4
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    • pp.383-390
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    • 1998
  • Bacillus subtilis DT134 was resistant to 50-fold higher concentration of cadmium ions (Cd2+) than cadmium-sensitive B. subtilis BD224 in Luria Broth (LB) medium. Minimal inhibition concentration test in LB agar plates also showed similar results. The elevated cadmium resistance of B. subtilis DT134 strongly suggested a possible existence of cadmium resistance gene in it. Southern blot with Staphylococcus aureus cadA gene fragment (757 bp NlaIV-XmnI cadA DNA fragment) as probe was carried out to test the existence and similarity of the gene. In high stringency condition, there was no detectable signal, but in low stringency, a strong signal specific to the cadA probe could be detected. These results strongly suggested that there was some similarity between total DNA of B. subtilis DT134 and S. aureus pl258 in terms of cadmium resistance gene and the resistance mechanism might be an efflux mechanism. The subsequent efflux experiment showed that the cadmium resistance mechanism of B. subtilis DT134 was also due to the efflux of cadmium.

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Complete Genome Sequence of Bacillus subtilis NIB353 Isolated from Nuruk

  • Jeong-Ah Yoon;Se-Young Kwun;Eun-Hee Park;Myoung-Dong Kim
    • Microbiology and Biotechnology Letters
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    • v.51 no.3
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    • pp.289-292
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    • 2023
  • Thermotolerant Bacillus subtilis NIB353 was isolated from Nuruk, a traditional Korean fermentation starter. The complete B. subtilis NIB353 genome sequence was obtained using MinION and Illumina (MiSeq) platforms. The B. subtilis NIB353 genome sequence was 4,247,447 bp with a GC content of 43%. The B. subtilis NIB353 strain exhibited orthologous average nucleotide identity values of 98.39% and 98.38% with B. subtilis 168 and B. subtilis ATCC6051a, respectively. The genome has been deposited in GenBank under the accession number NZ_CP089148.1.

Complete Genome of Bacillus subtilis subsp. subtilis KCTC 3135T and Variation in Cell Wall Genes of B. subtilis Strains

  • Ahn, Seonjoo;Jun, Sangmi;Ro, Hyun-Joo;Kim, Ju Han;Kim, Seil
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1760-1768
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    • 2018
  • The type strain Bacillus subtilis subsp. subtilis KCTC $3135^T$ was deeply sequenced and annotated, replacing a previous draft genome in this study. The tar and tag genes were involved in synthesizing wall teichoic acids (WTAs), and these genes and their products were previously regarded as the distinguishing difference between B. s. subtilis and B. s. spizizenii. However, a comparative genomic analysis of B. subtilis spp. revealed that both B. s. subtilis and B. s. spizizenii had various types of cell walls. These tar and tag operons were mutually exclusive and the tar genes from B. s. spizizenii were very similar to the genes from non-Bacillus bacteria, unlike the tag genes from B. s. subtilis. The results and previous studies suggest that the tar genes and the tag genes are not inherited after subspecies speciation. The phylogenetic tree based on whole genome sequences showed that each subspecies clearly formed a monophyletic group, while the tree based on tar genes showed that monophyletic groups were formed according to the cell wall type rather than the subspecies. These findings indicate that the tar genes and the presence of ribitol as a cell-wall constituent were not the distinguishing difference between the subspecies of B. subtilis and that the description of subspecies B. s. spizizenii should be updated.