• Title/Summary/Keyword: Bacillus identification

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Isolation, Identification , and Biodegradability of Phosphamidon-Degrading Bacteria (Phosphamidon 분해세균의 분리동정 및 생분해능)

  • 강양미;송홍규;안태석;허성남
    • Korean Journal of Microbiology
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    • v.35 no.1
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    • pp.61-64
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    • 1999
  • Organophosphorus inseclicide phosphamidon-degrading bacteria were isolated from agricultural soils and identified using Biolog microtiter assay. All Gram-positive degrading bacterial strains belong to genus Bacillus and many Gram-negative bacteria were rare soil species. Among them fast growing strains on phosphamidon-containing minimal medium were sclected and their biodegrading capability wcre measured. YD-17 which was identified as Capnocytophaga gingivalis showed the highest biodegradation rate. It could incrcase the removal of phosphamidon up to 52%. During the biodegradation continuous increase of amount of cell protein was observed, which indicated that phosphamidon was utilized as a carbon source for phosphamidon-degrading bacteria.

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A Thermostable Protease Produced from Bacillus sp. JE 375 Isolated from Korean Soil (한국의 토양으로부터 내열성 단백질 분해효소를 생산하는 Bacillus sp. JE 375의 선별)

  • Kim, Ji-Eun;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.38 no.3
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    • pp.419-426
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    • 2006
  • A thermophilic microorganism, strain JE 375, which produces a thermostable protease, was isolated from soil and compost in Korea. This gram-positive, rod-shaped, catalase positive, motility positive, and hemolysis ${\beta}$ containing organism was implicated in glucose fermentation, mannitol fermentation, xylose oxidation, aerobic activity and spore formation. The color of the colony was yellowish white. The temperature range for growth at pH 6.5 was between 55 and $70^{\circ}C$, with an optimum growth temperature of $65^{\circ}C$. This result confirmed the strain JE 375 as a thermophilic microorganism. The enzyme was produced aerobically at $65^{\circ}C$ during 20 hr in a medium (pH 6.5) containing 1% trypton. 1% maltose, 0.5% yeast extract and 1% NaCl. The 16S rDNA of strain JE 375 had 97.6% sequence similarity with the 16S rDNA of Bacillus caldoxyloyticus. On the basis of biochemical and physiological properties and phylogenetic analysis, we named the isolated strain as Bacillus sp. JE 375. The thermostable protease from Bacillus sp. JE 375 had been partially purified and characterized. The molecular weight of the enzyme was deduced from SDS-PAGE and gel chromatography as 55 kDa and its optimal temperature was $60^{\circ}C$. The enzyme showed its highest activity at pH 7.5 and was stable from pH 7.0 to 8.0.

Isolation and Identification of the Antagonistic Microorganisms Against Streptococous spp. Causing Dental Caries in Korean Soy Sauce (한국재래간장으로부터 구강질환 방제균의 선발 및 동정)

  • 엄수정;이여진;김진락;이은탁;김상달
    • Journal of Life Science
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    • v.13 no.4
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    • pp.535-540
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    • 2003
  • The antagonistic microorganisms against Streptococcus sanguis, S. salivarius and S. mutans causing the dental caries of oral diseases were isolated from Korean traditional soy sauce. Twenty five strains were isolated by pairing culture, paper disc culture and dual culture methods. The isolate NG 06 strain was observed with various cultural and physiological test, and $Biolog^{(R)}$ Bacterial Identification System. The strain was identified as Bacillus racemilacticus. The isolate NG 16 strain was confirmed to Gram-positive, rods, endospore production, utilization of melibiose, casein hydrolysis and starch hydrolysis. Also the second strain NG 16 was identified as $\beta$. amyloliquefaciens.

Availability of MADLDI-TOF MS for Identification of Gram Positive Bacilli Isolated from Blood Culture

  • Choi, Jin-Un;Kim, Sang-Ha;Hwang, Su-Jeong;Yu, Young-Bin;Kim, Sunghyun;Kim, Young-Kwon
    • Biomedical Science Letters
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    • v.24 no.2
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    • pp.108-115
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    • 2018
  • In the present study, results of the identification of Gram-positive bacilli (GPB) were analyzed by using the MALDI-TOF MS technique to score each 2-year blood culture at a university hospital. In addition, 16S rRNA sequence analyses and MALDI-TOF MS results are compared to targeting strains that had been isolated two or more times within the same patient, to evaluate the usefulness of MALDI-TOF MS in GPB identification. According to the cut-off (${\geq}1.7$) criteria, there were 410 (57.5%) reliable strains and 303 (42.5%) non-identified strains among the GPB identification results of 713 strains, using a microflex MALDI Biotyper (Bruker Daltonik GmbH, Bremen, Germany). The isolation appeared most often in the following order: Corynebacterium striatum, Bacillus cereus, Bacillus subtilis, Paenibacillus urinalis, and Listeria monocytogenes. Nearly three-fourths, 66 out of 89 (74.2%) of the strains for Corynebacterium striatum; 44 out of 60 (73.3%) strains for Bacillus cereus; and all (25 out of 25, 100%) Listeria monocytogenes strains were identified by their high scores of 2.0 or higher. Most (293 strains out of 303) non-identified strains were strains isolated only once and not significant as infectious bacilli. A total of 43 out of 50 (86.0%) strains matched and were able to be identified based on the 16 rRNA sequencing comparison results of strains that were isolated twice or more within the same patient and significant as infection bacilli. Non-matching among 5 out of 7 strains was not identified, even with MALDI-TOF MS. In conclusion, GPB can be identified in blood cultures using MALDI-TOF MS. This can be done accurately with ease, rapidly, and at a low cost. It is also thought to be helpful in GPB diagnosis and treatment.

Identification and Molecular Characterization of Three Isoforms of Iturin Produced by Endophytic Bacillus sp. CY22 (식물 내생균 Bacillus sp. CY22가 생성하는 iturin isoform의 분리 및 특성)

  • Cho, Soo-Jeong;Yun-Han-Dae
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.1005-1012
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    • 2005
  • Endophytic Bacillus sp. CY22 was previously isolated from the interior of balloon flower root and showed strong antifungal activity against phytopathogenic fungi such as Rhizoctonia solnni, Fusarium oxysporum, and Phythium ultimum. Many Bacillus strains produce antifungal compound such as iturin, fengycin, and mycosubtilin. We isolated and identified antifungal compound from cell supernatant of the endophytic strain. By the MALDI-TOF mass result, the antifungal compound was similar to the known antifungal lipopeptide iturin. It was found that the purified iturin had three isoforms with protonated masses of m/z 1,043.39, 1,057.42, and 1,071.42 and different structures in combination with $Na^{+}$ ion using MALDI-TOF MS. The ita22 gene, which transacylase gene is associated with production of antifungal iturin, had an open reading frame (ORF) of 1,200 bp encoding 400 amino acids. Results of deduced amino acids sequence homology search, Ita22 was homologous with FenF (BAB69697) of Bacillus subtilis 168.

Characterization of Bacillus licheniformis KJ-9 Isolated from Soil (토양으로부터 분리한 Bacillus licheniformis KJ 9의 특성)

  • Seo, Dong-Cheol;Ko, Jeong-Ae;Gal, Sang-Won;Lee, Sang-Won
    • Journal of Life Science
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    • v.20 no.3
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    • pp.403-410
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    • 2010
  • In order to produce high-quality fermenting composts, a microorganism was isolated from the natural world. The bacterium has not only in high enzyme activities but also had good antimicrobial activities against phytopathogenic microorganisms. Its cultivating characteristics were then investigated. Bacterium KJ-9, which contains high CMCase, protease and chitinase activities and excellent antimicrobial activities against phytopathogenic microorganisms, was separated from leaf mold and identified as Bacillus licheniformis by two methods: Bergey's Manual of Systematic Bacteriology and API 50 CHL Carbohydrate Test Kit (Bio Merieux, France) using an ATB (Automated Identification) computer system (Bio Merieux, France). Optimal medium for cultivation of B. licheniformis was 2% soluble starch as a carbon source, 0.5% yeast extract as a nitrogen source and 0.05% $MgSO_4{\cdot}7H_2O$. Optimal growth conditions of pH, temperature and shake speed were pH 7.0, $50^{\circ}C$ and 180 rpm, respectively. Culture broth of B. licheniformis KJ-9 cultured for 36~60 hr was effective in fungicidal activities against plant pathogens including Botrytis cinerea, Corynespora cassicola, Fusarium oxysporum, and Rhizoctonia solani.

Isolation and Identification of Cellulase-producing Microorganism, and Determination of Optimal Culture Condition (토양으로부터 Cellulose 분해효소를 생산하는 미생물의 분리, 동정 및 최적배양조건의 결정)

  • Hahm, Byoung-Kwon;Kim, Yoon-Keun;Yu, Ju-Hyun;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.29 no.5
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    • pp.1028-1032
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    • 1997
  • The strain No. 33, which produces cellulose-degrading enzyme, was isolated from soil. Yellow halo was identified when the culture supernatant of the strain was loaded onto agar plate containing 2.0% CMC using paper disc method. From scanning electron microscopic observation, the morphology of the stain was rod-shaped. For identification, several biochemical characteristics were tested, and this strain was identified to Bacillus sp. So, we named this strain as Bacillus sp. No. 33. The maximal growth was observed when the stain was cultured in the medium containing 1.0% glucose, 3.0% yeast extract, 0.5% $KH_2PO_4$, 0.02% $MgSO_4{\cdot}7H_2O$, pH 7.0 at $30^{\circ}C$ for 39 hours with shaking. The maximal enzyme production was accomplished using the medium containing 4.0% CMC, 2.0% yeast extract, 0.5% $KH_2PO_4$, 0.04% $MgSO_4{\cdot}7H_2O$, pH 7.0 at $30^{\circ}C$ for 42 hours with shaking.

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Identification of Food-Poisoning Bacteria (Bacillus cereus) and the Bacterial Toxin Genes for Application to Forensic Microbiology : A Case Report from National Forensic Service (법미생물 검사를 위한 식중독 세균(Bacillus cereus)의 동정 및 독소 유전자 검사법: 국립과학수사연구원 사례보고)

  • Cho, Yoonjung;Lee, Min Ho;Kim, Hyo Sook;Eom, Kiyoon;Kim, Min-Hee;Kim, Jong-Bae;Lee, Dong Sub
    • Journal of Science Criminal Investigation
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    • v.11 no.3
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    • pp.210-217
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    • 2017
  • In the forensic microbiology laboratories, microorganism analyses from food are requested. There have been several cases of Bacillus cereus isolated from the samples requested to the National Forensic Service. B. cereus is an important pathogenic bacterium which can cause food-borne outbreaks. Therefore, we isolated B. cereus from anchovy aekjeot recently requested for microbial examination and identified using MSId based on the 16S rDNA sequence and real-time PCR method. We also conducted PCR for detection of diarrheal toxin genes and an emetic toxin gene and found the presence of nheABC, bceT and entFM diarrheal toxin genes in the B. cereus isolate. There are several clinically important food-poisoning bacteria that should be noted during inspection. In particular, B. cereus can cause food poisoning even when cooked foods are ingested, because B. cereus forms endo-spore which confers strong environmental resistance and heat resistance to the bacteria, and the bacterial emetic toxin also has heat resistance. Here we highlight the importance to distinguish clinically important bacteria such as B. cereus from food specimens, and we expect this study will provide procedures for identification of B. cereus and detection of the bacterial toxin genes for future cases in the forensic microbiology laboratories.

Isolation and Charaterization of the Fibrinolytic Enzyme Producing Bacterium isolated from Naturally Fermented Chungkookjang (청국장에서 분리한 혈전용해효소 생산세균의 분리 및 동정)

  • Sohn Byung-Hee;Oh Kye-Heon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.7 no.3
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    • pp.476-482
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    • 2006
  • The aim of this work was to perform the screening and identification of the bacterium, MK-15 having the activity of fibrinolytic enzyme for the commercial use. Initially, strain MK-15 was enriched and isolated from naturally fermented soybean. Morphological and various physiological characteristics of the strain MK-15 was examined. The activity of fibrinolytic enzyme derived from supernatants of test culture MK-15 was performed by fibrin plate method for solid fibrinolytic activity. As the result, the fibrinolytic activity of MK-15 grown on the soybean media was about 2.7 times greater than that of plasmin used as stardard. 16S rRNA analyses revealed that strain MK-15 was 99.9% similar to Bacillus subtilis species cluster, and the bacterium was designated as Bacillus sp. MK-15. Strain MK-15 was registered in GenBank as [DQ163021].

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Structural Identification of $Siderophore_{AH18}$ from Bacillus subtilis AH18, a Biocontrol agent of Phytophthora Blight Disease in Red-pepper (Bacillus subtilis AH18의 고추역병 방제능과 $Siderophore_{AH18}$의 구조분석)

  • Woo, Sang-Min;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.326-335
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    • 2008
  • The siderophore ($siderophore_{AH18}$) of Bacillus subtilis AR18 was determined to be one of catechol type and purified by using Amberlite XAD-2, Sephadex LR-20 chromatography, and reversed-phase RPLC. The $Siderophore_{AH18}$ was identified bacillibactin with its structure by GC-MS, $^1H$-NMR, and $^{13}C$-NMR. $Siderophore_{AH18}$ (bacillibactin) had been confirmed its molecular weight of 883 and chemical structure of $(2,3-dihydroxybenzoate-glycine-threonine)_3$. Purified $siderophore_{AH18}$ showed strong biocontrol ability towards the spore of Phytophthora capsici on PDA and able to effectively suppress (55%) P. capsici causing red-pepper blight in the pot in vivo test.