• Title/Summary/Keyword: Bacillus sp. A1

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Purification and Characterization of β-Lactamase Secreted from Bacillus sp. J105 Strain having β-Lectam Antibiotics Resistance. ((β-lactam계 항생물질 저항성을 지닌 Bacillus sp. J105 균주로부터 분비되는 베타 락탐 분해효소의 정제 및 특성)

  • Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Min-Jeong;Lee, Young-Choon;Lee, Jai-Heon;Joo, Woo-Hong;Choi, Yung-Hyun;Lim, Hak-Seob;Kim, Jeong-In;Seo, Kwon-Il;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.6
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    • pp.845-851
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    • 2008
  • ${\beta}-Lactamase$, secreted from Bacillus sp. J105 strain was purified to a single band on SDS-PAGE by ammonium sulfate precipitation, ion exchange column chromatography and gel-filtration. The molecular weight of the purified enzyme was 31 kDa on SDS-PAGE and its isoelectric point was 7.35. Optimal pH and temperature for enzymatic reaction were 5 and $40^{\circ}C$, respectively. As a result of total amino acid composition analysis of the purified enzyme, Gly and Ala were occupied 14.1 and 13.3 mole %, respectively. Km and Vmax value of purified enzyme were 1.33 mM and 0.36 mM/ml using ampicillin as a substrate, respectively.

Screening of an Antagonistic Bacterium for Control of Red-pepper Anthracnose, Colletotrichum gloeosporioides (고추탄저병균 Colletotrichum gloeosporioides의 방제를 위한 길항 미생물의 분리 및 항진균 활성)

  • Park Sung-Min;Jung Hyuck-Jun;Yu Tae-Shick
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.420-426
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    • 2006
  • Bacillus sp. KMU-991 was isolated from Oslo city soils at Norway and shown a strong antifungal activity on red-pepper anthracnose, Colletotrichum gloeosporioides. Bacillus sp. KMU-991 produced a maximum level of antifungal substrate under aerobic incubation at $30^{\circ}C$, 180 rpm for 48 hours in TSB medium(initial pH 7.0) containing 1.0% mannitol and 1.0% ammonium chloride. Precipitate of culture broth by $30{\sim}60%$ ammonium sulfate precipitation exhibited strong antifungal activity against C. gloeosporioides KACC 40804. Butanol extract of cultured broth also shown fungal growth inhibitory activity against Fusarium oxysporum f. sp. radicus-lycopersici KACC 40537, Rhizoctonia solani AG-4 KACC 40142, Botrytis cinerea KACC 40573, Colletotrichum orbiculare KACC 40808, and Phytophthora cambivora KACC 40160 by agar diffusion method.

Thermostable $\alpha$-Amylase Production by Thermophilic Bacillus sp. TR-25 lsolated from Extreme Enviroment (극한환경에서 분리한 고온성 Bacillus sp. TR-25에 위한 내열성 $\alpha$-amylase의 생산)

  • 노석범;손홍주;이종근
    • Journal of Life Science
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    • v.7 no.1
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    • pp.30-38
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    • 1997
  • For screening thermostable $\alpha$-amylase from thermophiles, various samples from extreme environments such as hot spring and sewage near them, and compoat, wereexamined microbial growth in enrichment culture medium at 55$\circ$C on the assumption that enzymes from thermophiles are inevitable thermostable. One strain showing higher $\alpha$-amylase activity was pure cultured and designated as Bacillus sp. TR-25 from the results of morphological, cultural and physiological characteristics. The most important carbon sourses for the enzyme production were soluble starch, dextrin, potato starch and corn starch. Glucose and fructose had a catabolite repression on the enzyme production. The good nitrogen sources for the enzyme production were yeat extract, nutrient broth, tryptone, corn steep liquor and ammonium sulfate. The enzyme production was accelerated by addition of CaCl$_{2}$. $\cdot $ H$_{2}$O. The optimal medium composition for the enzyme production was soluble starch 2.0%, yeast extract 0.55, CaCl$_{2}$ $\cdot $ 2H$_{2}$O 0.015, Tween 80 0.001%, pH8.0, respectively. In jar fermenter culture, this strain shows a rapid growth and required cheaper carbon and nitrogen source. These properties are very useful to fermentation industry. The $\alpha$-amylase of this strain demonstrated a maximum activity at 80$\circ$C, pH 5.0, respectively. And calcium ion did not improve thermostability of the enzyme. At 10$0^{\circ}C$, this enzyme has 235 of relative activity. Transformation was carried out by thermophilic Bacillus sp. TR-25 genomic DNA. As a result, the transformant has increased thermostable $\alpha$-amylase activity.

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Isolation and Identification of Microorganism with Potent Fibrinolytic Activity from Korean Traditional Deonjang (재래식 된장으로부터 혈전용해활성을 나타내는 세균의 분리 및 동정)

  • Hyun Kwang-Wook;Lee Jong-Soo;Ham Jung-Hee;Choi Shin-Yang
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.24-28
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    • 2005
  • A bacterial strain D­1 found to have potent fibrinolytic activity was isolated from Korean traditional Doenjang. It was identified as Bacillus sp. based on its 16S rRNA sequence analysis, morphological and physiological characteristics.

Screening of Antifungal Bacillus spp. against Alternaria Blight Pathogen (Alternaria panax) and Anthracnose Pathogen (Colletotrichum gloeosporioides) of Ginseng (인삼 점무늬병균(Alternaria panax)과 탄저병균(Colletotrichum gloeosporioides)에 대한 길항미생물 Bacillus spp. 선발)

  • Lee, Hye-Jin;Park, Kee-Choon;Lee, Seung-Ho;Bang, Kyong-Hwan;Park, Hong-Woo;Hyun, Dong-Yun;Kang, Seung-Weon;Cha, Seon-Woo;Chung, Ill-Min
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.5
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    • pp.339-344
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    • 2012
  • This study was carried out to research microorganisms having the antifungal activity against ginseng Alternaria blight pathogen Alternaria panax and ginseng anthracnose pathogen Colletotrichum gloeosporioides. Eleven Bacillus strains. were isolated from Korean traditional soybean paste and Kimchi. Among the 11 isolates, DJ5, KC1, KC2 and KC4 showing antagonistic activity on the mycelial growth of A. panax and C. gloeosporioides in pairing culture were finally selected as the antagonistic microorganisms. Based on 16s rRNA sequence and phylogenetic tree analysis, they were identified as Bacillus spp.. The selected microorganisms were investigated antagonistic activity by measured leaf-segment colonization in pot test. When Bacillus sp. were injected after A. panax treatment, KC1, KC2 and KC4 showed similar effect to chemical pesticides treated control. To measure preventive effect of Bacillus sp, antagonistic microorganisms were injected and C. gloeosporioides was treated in pot. When measuring the effectiveness for the prevention of Anthracnose, All Bacillus spp. showed approximately 83~90 % degree of superior preventive effect. In general, The four Bacillus spp. isolated from Korean traditional fermented foods showed therapeutic effect of Alternaria blight and preventive effect of Anthracnose.

The Effective Isolation of a Mosquitocidal Bacteria, Bacillus thuringiensis Subsp. israelensis (모기 살충성 세균 B. thuringiensis subsp. israelensis의 효과적인 분리 방법)

  • 김광현;이광배;신두만
    • Journal of environmental and Sanitary engineering
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    • v.13 no.2
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    • pp.34-39
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    • 1998
  • For more convenient and rapidly isolation of Bacillus thuringiensis subsp. israelensis(Bti), 1) heat treatment spore forming bacteria, 2) growth in enrichment culture media for Bacillus sp. and 3) selection of bacteria producing a lecithinase for Bacillus thuringiensis subsp. israelensis, were performed. Spore forming bacteria were counted 4.8 $\times $ 10$^{8}$cells/g soil on NAPGCY media, 9.2 $\times $ 10$^{7}$cells/g soil on NA media, and 3.6 $\times $ 10$^{8}$cells/g soil on NAAC media, respectively. Bacteria producing only a lecithinase were reached at 25.2% on medium contained egg york, bacteria only producing a delta-endotoxin were reached at 23.2% by phase contrast microscope, and bacteria producing a lecithinase & a delta-endotoxin simultaneously were reached at 13.7%. Bacillus thuringiensis which producing a lecithinase and a delta-endotoxin simultaneously among bacteria producing a lecithinase, were reached at 56.5%; A half of Bacillus thuringiensis was produced a delta-endotoxin, but not produced a lecithinase. Among 8 isolates of Bacillus thuringiensis, two strain of Bti which has a mosquito-cidal toxin, were detected by PCR using a specific primer of $\delta $-endotoxin gene from Bti.

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Optimization of Laccase Production from Bacillus sp. PK4 through Statistical Design of Experiments

  • Rajeswari, Murugesan;Bhuvaneswari, Vembu
    • Microbiology and Biotechnology Letters
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    • v.45 no.4
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    • pp.330-342
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    • 2017
  • Statistical design of experiments was employed to optimize the media composition for the production of laccase from Bacillus sp. PK4. In order to find the key ingredients for the best yield of enzyme production from the selected eleven variables viz yeast extract, glucose, zinc sulphate, copper sulphate, potassium chloride, magnesium sulphate, calcium chloride, ferrous sulphate, sodium chloride, potassium dihydrogen phosphate ($KH_2PO_4$) and dipotassium hydrogen phosphate ($K_2HPO_4$), Plackett-Burman design was applied. The $MgSO_4$, $FeSO_4$, and $CuSO_4$ showed positive estimate, and their concentration optimized further. The steepest ascent method and Box-Behnken method revealed that 1.5 mM $MgSO_4$, 0.33 g/l $FeSO_4$ and 1.41 mM $CuSO_4$ were optimal for the laccase production by Bacillus sp. PK4. This optimization strategy leads to enhancement of laccase production from 2.13 U/ml to 40.79 U/ml. Agro-wastes residues replace the carbon source glucose in the optimized media namely sugarcane bagasse, wheat bran, rice husk, and groundnut shell, among these groundnut shells (117 U/ml) was found to enhance the laccase production significantly. The laccase produced by Bacillus sp. PK4 was found to have the potential to degrade persistent organic pollutant benzo[a]pyrene.

An Antibiotic against Bacillus amyloliquefaciens from Streptomyces sp. KM-48 (Streptomyces 속 KM-48 균주가 생성하는 Bacillus amyloliquefaciens 에 대한 항생물질)

  • Lee, Sang-Man;Song, Bang-Ho
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.39-45
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    • 1983
  • One of Streptomyces sp. was screened as an antibiotic producing strain against Gram positive bacteria specifically from about one hundred Actinomycetes collections. The isolated KM-48 strain, indexed in our collection, was secreted an anbitiotic in nutrient broth culture. The active principle was extracted with acetone from the lyophilized solid of cultured filtrate. The nonadsorbed elute in aluminium oxide was isolated as a purified homogeneity which revealed a sing1e spot under ultra-violet beam on avicel thin layer chromatography. The isolated antibiotic was sensitive specifically against the Gram positive bacteria and fungi. About fifty percent inhibition of the control growth on Bacillus amyloliquefaciens was 18$\mu\textrm{g}$/$m\ell$. The antibiotic was lavile against thermotreatment and stabled in acidic condition remarkably. The KM-48 was identified as a similar strain to Streptomyces tsusimaensis on its morphological and cultural characteristics.

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Advanced Wastewater Treatment using Sequencing Batch Reactor on ship's sewage (연속 회분식 공정(SBR)을 이용한 선박 오·폐수의 고도처리)

  • Park Sang-Ho;Kim In-Soo
    • Journal of Navigation and Port Research
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    • v.29 no.5 s.101
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    • pp.475-480
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    • 2005
  • Lab scale experimental study was carried out for SBR process, to investigate the effects of influent ship sewage organic compound removal and Bacillus sp. state on design parameters. This process was able to remove nitrogen and phosphorus as well as organic matter efficiently. More than $92.0\%$ of chemical oxygen demand(COD) were removed. In addition, about $84.0\%$ of total nitrogen (T-N) was reduced. The total phosphorus(T-P) reduction averaged $93\%$. The performance load of SBR process was shown to be $0.095kg{\cdot}TOC/m3{\cdot}day$. The pH was decreased from 8.1 to 7.0 within 30 min and increased to 7.3 at the end of anoxic stage, and these phenomena were explained. The sludge produced in the SBR process is characterized by low generation rate (about $0.36kg{\cdot}MLSS/kg{\cdot}TOC$) and excellent settleability. The number of Bacillus sp. in the SBR was $24.2\%$, indicating that Bacillus sp. was a predominant species in the reactor.

Characterization of antimicrobial proteins produced by Bacillus sp. N32 (Bacillus sp. N32 균주가 생산하는 항균 단백질 특성)

  • Lee, Mi-Hye;Park, In-Cheol;Yeo, Yun-Soo;Kim, Soo-Jin;Yoon, Sang-Hong;Lee, Suk-Chan;Chung, Tae-Young;Koo, Bon-Sung
    • The Korean Journal of Pesticide Science
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    • v.10 no.1
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    • pp.56-65
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    • 2006
  • An antagonistic bacterial isolate, that inhibits the growth of plant pathogens, was selected and identified from 5,000 isolates screened from the rhizosphere of various crop plants. An isolate Bacillus sp. N32, tested against Colletotrichum gloeosporioides causing anthracnose disease in hot pepper, produced both a heat resistant antifungal protein and a heat sensitive antifungal protein. The heat resistant protein was partially purified by Ammonium sulfate fractionation and gel filtration chromatography. The bioautography showed that the proteins possessed high antifungal activity. The biosynthetic gene cluster responsible for the heat resistant antifungal protein was cloned from cosmid library using DNA probe obtained from PCR product with the primers targeting the conserved nucleotide sequence of the synthetic genes reported earlier, Most of the clones obtained showed higher homology to fengycin antibiotic synthetic gene family reported earlier. On the other hand, the heat sensitive protein was isolated from SDS-PAGE and electroblotting to determine the N-terminal amino acid sequences. The heat sensitive antifungal protein gene was cloned from the ${\lambda}-ZAP$ libraries using a DNA probe based on the N-terminal amino acid sequences of the heat sensitive protein. We are contemplating to clone and sequence the whole gene cluster encoding the heat sensitive protein for further analysis.