• Title/Summary/Keyword: Bacterial Isolates

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Alcaligenes eutrophus 균주의 성장과 Poly-Beta-hydroxybutyrate 합성에 미치는 포도당과 암모늄농도의 영향

  • 이용우;유영제
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.607-612
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    • 1990
  • The biodegradation of Aroclor 1242 was investigated by the mixed cultivation of the natural bacterial isolates and a genetically engineered microorganism (GEM). The natural strain of MS-1003 degraded the Aroclor 1242 through the ortho-cleavage pathway, while the other strains through the meta-cleavage pathway. When the MS-1003 strain was additionally inoculated into the 1 day culture of the DJ-26 strain and then cultivated for 2 days, the Aroclor was degraded up to 86% and resulted in increase of the meta-cleavage product. But in the MS-1003 culture inoculated with the DJ-26, degradation of the Aroclor was limited to the level of each pure culture. By the mixed cultivation of the DJ-26 strain together with the DJ-12 or its GEM strain of DF-10, which degrades the Aroclor through the meta-cleavage pathway, degradation of the Aroclor as well as production of the meta-cleavage compound were lower than those of each pure culture. The degradation of Aroclor 1242 by the GEM strain was not improved over the parental strain. Therefore, a form of cometaboiism of Aroclor 1242 was found in the mixed culture of the DJ-26 and MS-1003 strains which degrade the Aroclor through the different metabolic pathway, but in the mixed culture of the DJ-26 and DJ-12 strains degrading Aroclor 1242 through the same pathway, a kind of competetion for the substrate was observed.

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Use of Bioluminescent Indicator Acinetobacter Bacterium for Screening and Characterization of Active Antimicrobial Agents

  • Haleem Abd-El;A.M. Desouky;Zaki Sahar A.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1706-1712
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    • 2006
  • Because of the need for new antimicrobial substances with novel mechanisms of action, we report here the use of an Acinetobacter reporter system for high-throughput screening of active antimicrobial agents. The bioreporter Acinetobacter strain DF4/PUTK2 carrying luciferase genes luxCDABE was chosen because of its ecological importance and it is widespread in nature. This bioreporter is genetically engineered to emit light constitutively that can be measured in real time by luminometry. Hence, this reporter system was employed to determine the bacteriostatic actions of spent-culture supernatants derived from twelve bacterial isolates. Out of the results, the strongest bioluminescence inhibitory effect of the supernatants was recorded with Bacillus cereus strain BAC (S5). Subsequently, ethyl acetate extracts of extracellular products of strain BAC (S5) were separated by a thin-layer chromatography (TLC). Based on the bioluminescence inhibitory assay, three fractions were found to have antimicrobial activity. One fraction (C) having the strongest antimicrobial activity was further purified using TLC and characterized by IR, $^1H$ NMR, mass spectrometry, SDS-PAGE, and amino acid composition analysis. The results predicted the presence of 2-pyrrolidone-S-carboxylic acid (PCA) and the octadeconic-acid-like fatty acid. Fraction C also demonstrated a broad inhibitory activity on several Gram-negative and Gram-positive bacteria. In conclusion, the Acinetobacter reporter system shows great potential to be a reliable, sensitive, and real-time indicator of the bacteriostatic actions of the antimicrobial agents.

Temperature Effects on Korean Entomopathogenic Nematodes, Steinernema glaseri and S. longicaudum, and their Symbiotic Bacteria

  • Hang Dao Thi;Choo, Ho-Yul;Lee, Dong-Woon;Lee, Sang-Myeong;Kaya Harry K.;Park, Chung-Gyoo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.420-427
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    • 2007
  • We investigated the temperature effects on the virulence, development, reproduction, and otility of two Korean isolates of entomopathogenic nematodes, Steinernema glaseri Dongrae strain and S. longicaudum Nonsan strain. In addition, we studied the growth and virulence of their respective symbiotic bacterium, Xenorhabdus poinarii for S. glaseri and Xenorhabdus sp. for S. longicaudum, in an insect host at different temperatures. Insects infected with the nematode-bacterium complex or the symbiotic bacterium was placed at $13^{\circ}C,\;18^{\circ}C,\;24^{\circ}C,\;30^{\circ}C,\;or\;35^{\circ}C$ in the dark and the various parameters were monitored. Both nematode species caused mortality at all temperatures tested, with higher mortalities occurring at temperatures between $24^{\circ}C\;and\;30^{\circ}C$. However, S. longicaudum was better adapted to cold temperatures and caused higher mortality at $18^{\circ}C$ than S. glaseri. Both nematode species developed to adult at all temperatures, but no progeny production occurred at $13^{\circ}C\;or\;35^{\circ}C$. For S. glaseri, nematode progeny production was best at inocula levels above 20 infective juveniles/host at $24^{\circ}C\;and\;30^{\circ}C$, but for S. longicaudum, progeny production was generally better at $24^{\circ}C$. Steinernema glaseri showed the greatest motility at $30^{\circ}C$, whereas S. longicaudum showed good motility at $24^{\circ}C\;and\;30^{\circ}C$. Both bacterial species grew at all tested temperatures, but Xenorhabdus sp. was more virulent at low temperatures $(13^{\circ}C\;and\;18^{\circ}C)$ than X. poinarii.

The New LM-PCR/Shifter Method for the Genotyping of Microorganisms Based on the Use of a Class IIS Restriction Enzyme and Ligation-Mediated PCR

  • Krawczyk, Beata;Leibner-Ciszak, Justyna;Stojowska, Karolina;Kur, Jozef
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1336-1344
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    • 2011
  • This study details and examines a novel ligation-mediated polymerase chain reaction (LM-PCR) method. Named the LM-PCR/Shifter, it relies on the use of a Class IIS restriction enzyme giving restriction fragments with different 4-base, 5' overhangs, this being the Shifter, and the ligation of appropriate oligonucleotide adapters. A sequence of 4-base, 5' overhangs of the adapter and a 4-base sequence of the 3' end of the primer(s) determine a subset of the genomic restriction fragments, which are amplified by PCR. The method permits the differentiation of bacterial species strains on the basis of the different DNA band patterns obtained after electrophoresis in polyacrylamide gels stained with ethidium bromide and visualized in UV light. The usefulness of the LM-PCR/Shifter method for genotyping is analyzed by a comparison with the restriction endonuclease analysis of chromosomal DNA by the pulsed-field gel electrophoresis (REA-PFGE) and PCR melting profile (PCR MP) methods for isolates of clinical origin. The clustering of the LM-PCR/Shifter fingerprinting data matched those of the REA-PFGE and PCR MP methods. We found that the LM-PCR/Shifter is rapid, and offers good discriminatory power and excellent reproducibility, making it a method that may be effectively applied in epidemiological studies.

Characterization of a Phenazine and Hexanoyl Homoserine Lactone Producing Pseudomonas aurantiaca Strain PB-St2, Isolated from Sugarcane Stem

  • Mehnaz, Samina;Baig, Deeba Noreen;Jamil, Farrukh;Weselowski, Brian;Lazarovits, George
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1688-1694
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    • 2009
  • A novel strain of fluorescent pseudomonad (PB-St2) was isolated from surface-sterilized stems of sugarcane grown in Pakistan. The bacterium was identified as Pseudomonas aurantiaca on the basis of 16S rRNA gene sequence analysis and results from physiological and biochemical characteristics carried out with API50 CH and QTS 24 bacterial identification kits. Assays using substrate-specific media for enzymes revealed lipase and protease activities but cellulase, chitinase, or pectinase were not detected. The bacterium was unable to solubilize phosphate or produce indole acetic acid. However, it did produce HCN, siderophores, and homoserine lactones. In dual culture assays on agar, the bacterium showed antifungal activity against an important pathogen of sugarcane in Pakistan, namely Colletotrichum falcatum, as well as for pathogenic isolates of Fusarium oxysporium and F. lateritium but not against F. solani. The antifungal metabolites were identified using thin-layer chromatography, UV spectra, and MALDI-TOFF spectra and shown to be phenazine-1-carboxylic acid (PCA), 2-hydroxyphenazine (2-OH-PHZ), and N-hexanoyl homoserine lactone (HHL) (assessed using only TLC data). The capacity of this bacterium to produce HCN and 2-OH-PHZ, as well as to inhibit the growth of C. falcatum, has not been previously reported.

Cr(VI) Resistance and Removal by Indigenous Bacteria Isolated from Chromium-Contaminated Soil

  • Long, Dongyan;Tang, Xianjin;Cai, Kuan;Chen, Guangcun;Shen, Chaofeng;Shi, Jiyan;Chen, Linggui;Chen, Yingxu
    • Journal of Microbiology and Biotechnology
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    • v.23 no.8
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    • pp.1123-1132
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    • 2013
  • The removal of toxic Cr(VI) by microorganisms is a promising approach for Cr(VI) pollution remediation. In the present study, four indigenous bacteria, named LY1, LY2, LY6, and LY7, were isolated from Cr(VI)-contaminated soil. Among the four Cr(VI)-resistant isolates, strain LY6 displayed the highest Cr(VI)-removing ability, with 100 mg/l Cr(VI) being completely removed within 144 h. It could effectively remove Cr(VI) over a wide pH range from 5.5 to 9.5, with the optimal pH of 8.5. The amount of Cr(VI) removed increased with initial Cr(VI) concentration. Data from the time-course analysis of Cr(VI) removal by strain LY6 followed first-order kinetics. Based on the 16S rRNA gene sequence, strain LY6 was identified as Pseudochrobactrum asaccharolyticum, a species that had never been reported for Cr(VI) removal before. Transmission electron microscopy and energy dispersive X-ray spectroscopy analysis further confirmed that strain LY6 could accumulate chromium within the cell while conducting Cr(VI) removal. The results suggested that the indigenous bacterial strain LY6 would be a new candidate for potential application in Cr(VI) pollution bioremediation.

Statistical Analysis of Antimicrobial Susceptibility Tested on Various Clinical Isolates of Bacteria (임상검체에서 분리된 세균의 항생제 감수성에 관한 통계적 고찰)

  • Bae, Eun-Kyung;Jeon, Chang-Ho;Hong, Seok-Il;Kim, Chung-Sook
    • Journal of Yeungnam Medical Science
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    • v.3 no.1
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    • pp.185-192
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    • 1986
  • Antimicrobial susceptibility of the bacterial strains isolated from clinical specimens during the period from June, 1983 to June, 1986 in Yeungnam Medical Center was studied and the following results were obtained. 1. Staphylococcus aureus was highly susceptible to cephalothin and its susceptibility to methicillin was gradually reduced. 2. Streptococcus strains except enterococcus were generally susceptible to penicillin, while most enterococci were suscesceptible to only ampicillin. 3. Gram-negative rods including Escherichia coli were highly susceptible to amikacin and tobramycin. 4. Serratia were generally less susceptible to the amtimicrobials tested than other Enterobacteriaceae. Among them, Serratia marcescens showed the highest susceptibility to amikacin and chloramphenicol. 5. Pseudomonas aeruginosa revealed the highest susceptibility to amikacin and tobramycin and moderate susceptibility to carbenicillin and gentamycin. 6. Acinetobacter calcoaceticus revealed low susceptibility to most antimicrobials tested, showing only 30% susceptibility to amikacin, tobramycin and gentamycin in 1986.

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Calcium Carbonate Precipitation by Bacillus and Sporosarcina Strains Isolated from Concrete and Analysis of the Bacterial Community of Concrete

  • Kim, Hyun Jung;Eom, Hyo Jung;Park, Chulwoo;Jung, Jaejoon;Shin, Bora;Kim, Wook;Chung, Namhyun;Choi, In-Geol;Park, Woojun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.540-548
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    • 2016
  • Microbially induced calcium carbonate precipitation (CCP) is a long-standing but re-emerging environmental engineering process for production of self-healing concrete, bioremediation, and long-term storage of CO2. CCP-capable bacteria, two Bacillus strains (JH3 and JH7) and one Sporosarcina strain (HYO08), were isolated from two samples of concrete and characterized phylogenetically. Calcium carbonate crystals precipitated by the three strains were morphologically distinct according to field emission scanning electron microscopy. Energy dispersive X-ray spectrometry mapping confirmed biomineralization via extracellular calcium carbonate production. The three strains differed in their physiological characteristics: growth at alkali pH and high NaCl concentrations, and urease activity. Sporosarcina sp. HYO08 and Bacillus sp. JH7 were more alkali- and halotolerant, respectively. Analysis of the community from the same concrete samples using barcoded pyrosequencing revealed that the relative abundance of Bacillus and Sporosarcina species was low, which indicated low culturability of other dominant bacteria. This study suggests that calcium carbonate crystals with different properties can be produced by various CCP-capable strains, and other novel isolates await discovery.

Nitrogen Fixation Screening and Plant Growth Assessment for Urban Greening (도시 녹화를 위한 질소고정 균 선별 및 식물 생장 평가)

  • Jeong, Sun Hwan;Lee, Sang Seob
    • Microbiology and Biotechnology Letters
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    • v.46 no.2
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    • pp.154-161
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    • 2018
  • Currently, urban greening projects and research are attracting attention as a way to mitigate urban heat island phenomenon. In this study, nitrogen fixative bacteria were isolated and their effects on plant growth were confirmed. First, enrichment was performed in a nitrogen-free medium to isolate the nitrogen-fixing bacteria, and the colony showing high growth in a medium with limited nitrogen source was isolated and purified. Separated bacterial isolates were reduced by more than 90% acetylene by ARA and indirectly confirmed the activity of nitrogenase by ethylene production. Cedecea sp. MK7 and Enterobacter sp. Y8 with confirmed reproducibility were selected as nitrogen fixative bacteria. Nitrogen fixing bacteria were applied to the growth of perennial rye grass, and it was found that the dry weight increased to 34.80 mg (186.60%) compared with the control with 18.65 mg dry weight. After plant growth, microbial community analysis of soil applied by bacteria showed similarity to the control group. Therefore, in this study, it is expected that the efficiency will be increased if plant growth is promoted by using nitrogen fixing bacteria in urban greenery system.

Yellow-colored mats in Jeju Island lava tubes

  • Kim, Jong-Shik;Kim, Dae-Shin;Lee, Keun Chul;Kim, Yong-Hwan;Ahn, Ung-San;Lee, Mi-Kyung;Lee, Jung-Sook
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.4
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    • pp.1338-1348
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    • 2019
  • The Geomunoreum Lava Tube System, declared as a UNESCO World Heritage Site, has a unique natural ecosystem. The information available about this ecosystem, which contains lava caves with secondary carbonate speleothems, is sparse. Hence, extensive research is warranted for establishing a conservation standard. We commenced microbial research on the system and have been studying the microorganisms coating the lava tube wall to acquire fundamental information for understanding the lava cave ecology of Jeju Island. Samples were collected from yellow-colored walls in six caves that are part of the system-the Bengdwi, Utsanjeon, Bukoreum, Manjang, Gimnyeong, and Yongcheon caves. This study focused on yellow walls as it is the most easily distinguished color. According to previous studies, the color of cave walls is attributed to microorganisms or their components. To determine whether the yellow mats from the Jeju lava tube walls are caused by microorganisms, we examined samples at the microscopic scale, by staining mats and analyzing bacterial isolates from glitter particles. As a result, we found that the yellow walls of lava tubes are comprised of microbial mats.