• Title/Summary/Keyword: Gram-positive bacterium

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Isolation and Characterization of a Nematicidal Bacillus thuringiensis strain 108 (항선충성 Bacillus thuringiensis 108균주의 분리와 특성)

  • Lee, Jae-Hun;Ryu, Eun-Ju;Kim, Kwang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.250-254
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    • 2007
  • Bacillus thringiensis strain 108 was isolated from soil and had nematicidal activity against second stage juvenile of plant root-knot nematode, Meloidogyne incognita. The strain 108, a rod shape, spore forming and Gram positive bacterium, produced lecithinase, catalase, and ${\delta}$-endotoxin. The strain 108 belongs to H serotype 3, Bacillus thuringiensis var. kurstaki. A nematicidal substance of the strain 108 was partially purified on Sephadex G-25 gel filtration, activated carbon adsorption, silica gel adsorption, and Sephadex G-10 gel filtration. $LC_{90}$ of the partially purified substance against M incognita was $1.2\;{\mu}g/ml$. The nematicidal substance was stable by heat treatment at $100^{\circ}C$ for 1hr, but was perfectly lost nematicidal activity after autoclave ($110^{\circ}C$, 30 min).

Involvement of Lipopolysaccharide of Bradyrhizobium japonicum in Metal Binding

  • Oh, Eun-Taex;Yun, Hyun-Shik;Heo, Tae-Ryeon;Koh, Sung-Cheol;Oh, Kye-Heon;So, Jae-Seong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.296-300
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    • 2002
  • Bacterial cell surface components are the major factors responsible for pathogenesis and bioremediation. In particular, the surface of a Gram-negative bacterium cell has a variety of components compared to that of a Gram-positive cell. In our previous study, we isolated an isogenic mutant of Bradyrhizobium japonicum, which exhibited altered cell surface characteristics, including an increased hydrophobicity. Polyacrylamide gel electrophoretic analysis of the lipopolysaccharide (LPS) in the mutant demonstrated that the O-polysaccharide part was completely absent. Meanwhile, a gel permeation chromatographic analysis of the exopolysaccharide (EPS) in the mutant demonstrated that it was unaltered. Since LPSs are known to have several anion groups that interact with various cation groups and metal ions, the mutant provided an opportunity to examine the direct role of LPS in metal binding by B. japonicum. Using atomic absorption spectrophotometry, it was clearly demonstrated that LPS was involved in metal binding. The binding capacity of the LPS mutant to various metal ions $(Cd^{2+},\;Cu^{2+},\;Pb^{2+},\;and\;Zn^{2+})$ was 50-70% lower than that of the wild-type strain. Also, through an EPS analysis and desorption experiment, it was found that EPS and centrifugal force had no effect on the metal binding. Accordingly, it would appear that LPS molecules on B. japonicum effect the properties, which precipitate more distinctly metal-rich mineral phase.

Chemosystematic and Phenotypic Characterization of Gram-positive Bacteria from Coastal Seawater, Korea (한국 근해에서 분리한 그람양성 세균의 화학 분류학적 및 표현형적 특성)

  • 전정훈;박진숙
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.167-172
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    • 2000
  • Twenty-five halotolerant gram-positlve bacteria were isolated from the coastal seawater 01 Cheju Island and Incheon J&yakdo Chemosystematic and phenotypic characteristics were used to iuvestigate the taxonomic position of these bacteria. According to their chemosystematic characteristics, the twenty-tive isolates were divided into 4 groups. Group 1 bacteria possesed 40.1 to 49.9 inol% m DNA G+C content, menaquinone-7 as a major quinone, and meso-Alpm as a diamino acid of peptidoglycan. Group 1 tam were identified as Bacilluspumilis, Bacillus lichenifbrrizis, Bacillus megaterium, Bncill~rsubtilis. Group 2 bacteria possessed 63.9 to 66.4 mol% and MK-8. They were all in the genus Arth~obaclm-. Group 3 bacteria possessed 31.0 to 37.6 mol% and MK-7. They were identified as Staphylococcus haeniol.viicvs, Siaph~~lococc~is sapropl~j~ticns, and Siaphylococcus intermediru.. Group 4 bacterium possessed 72.0mol% and MK-8 and was identified as ~Lficrococcus ltdtezm. Bacillus species accounted for 68% of h e total isolates.

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NADP+-Dependent Dehydrogenase SCO3486 and Cycloisomerase SCO3480: Key Enzymes for 3,6-Anhydro-ʟ-Galactose Catabolism in Streptomyces coelicolor A3(2)

  • Tsevelkhorloo, Maral;Kim, Sang Hoon;Kang, Dae-Kyung;Lee, Chang-Ro;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.756-763
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    • 2021
  • Agarose is a linear polysaccharide composed of ᴅ-galactose and 3,6-anhydro-ʟ-galactose (AHG). It is a major component of the red algal cell wall and is gaining attention as an abundant marine biomass. However, the inability to ferment AHG is considered an obstacle in the large-scale use of agarose and could be addressed by understanding AHG catabolism in agarolytic microorganisms. Since AHG catabolism was uniquely confirmed in Vibrio sp. EJY3, a gram-negative marine bacterial species, we investigated AHG metabolism in Streptomyces coelicolor A3(2), an agarolytic gram-positive soil bacterium. Based on genomic data, the SCO3486 protein (492 amino acids) and the SCO3480 protein (361 amino acids) of S. coelicolor A3(2) showed identity with H2IFE7.1 (40% identity) encoding AHG dehydrogenase and H2IFX0.1 (42% identity) encoding 3,6-anhydro-ʟ-galactonate cycloisomerase, respectively, which are involved in the initial catabolism of AHG in Vibrio sp. EJY3. Thin layer chromatography and mass spectrometry of the bioconversion products catalyzed by recombinant SCO3486 and SCO3480 proteins, revealed that SCO3486 is an AHG dehydrogenase that oxidizes AHG to 3,6-anhydro-ʟ-galactonate, and SCO3480 is a 3,6-anhydro-ʟ-galactonate cycloisomerase that converts 3,6-anhydro-ʟ-galactonate to 2-keto-3-deoxygalactonate. SCO3486 showed maximum activity at pH 6.0 at 50℃, increased activity in the presence of iron ions, and activity against various aldehyde substrates, which is quite distinct from AHG-specific H2IFE7.1 in Vibrio sp. EJY3. Therefore, the catabolic pathway of AHG seems to be similar in most agar-degrading microorganisms, but the enzymes involved appear to be very diverse.

Impact of UV-C Irradiation on Bacterial Disinfection in a Drinking Water Purification System

  • Hyun-Joong Kim;Hee-Won Yoon;Min-A Lee;Young-Hoon Kim;Chang Joo Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.1
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    • pp.106-113
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    • 2023
  • The supply of microbiological risk-free water is essential to keep food safety and public hygiene. And removal, inactivation, and destruction of microorganisms in drinking water are key for ensuring safety in the food industry. Ultraviolet-C (UV-C) irradiation is an attractive method for efficient disinfection of water without generating toxicity and adversely affecting human health. In this study, the disinfection efficiencies of UV-C irradiation on Shigella flexneri (Gram negative) and Listeria monocytogenes (Gram positive) at various concentrations in drinking water were evaluated using a water purifier. Their morphological and physiological characteristics after UV-C irradiation were observed using fluorescence microscopy and flow cytometry combined with live/dead staining. UV-C irradiation (254 nm wavelength, irradiation dose: 40 mJ/cm2) at a water flow velocity of 3.4 L/min showed disinfection ability on both bacteria up to 108 CFU/4 L. And flow cytometric analysis showed different physiological shift between S. flexneri and L. monocytogenes after UV-C irradiation, but no significant shift of morphology in both bacteria. In addition, each bacterium revealed different characteristics with time-course observation after UV-C irradiation: L. monocytogenes dramatically changed its physiological feature and seemed to reach maximum damage at 4 h and then recovered, whereas S. flexneri seemed to gradually die over time. This study revealed that UV-C irradiation of water purifiers is effective in disinfecting microbial contaminants in drinking water and provides basic information on bacterial features/responses after UV-C irradiation.

Bacillus anthracis Spores Influence ATP Synthase Activity in Murine Macrophages

  • Seo, Gwi-Moon;Jung, Kyoung-Hwa;Kim, Seong-Joo;Kim, Ji-Cheon;Yoon, Jang-Won;Oh, Kwang-Keun;Lee, Jung-Ho;Chai, Young-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.778-783
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    • 2008
  • Anthrax is an infectious disease caused by toxigenic strains of the Gram-positive bacterium Bacillus anthracis. To identify the mitochondrial proteins that are expressed differently in murine macrophages infected with spores of B. anthracis Sterne, proteomic and MALDI-TOF/MS analyses of uninfected and infected macrophages were conducted. As a result, 13 mitochondrial proteins with different expression patterns were discovered in the infected murine macrophages, and some were identified as ATP5b, NIAP-5, ras-related GTP binding protein B isoform CRAa, along with several unnamed proteins. Among these proteins, ATP5b is related to energy production and cytoskeletal rearrangement, whereas NIAP-5 causes apoptosis of host cells due to binding with caspase-9. Therefore, this paper focused on ATP5b, which was found to be down regulated following infection. The downregulated ATP5b also reduced ATP production in the murine macrophages infected with B. anthracis spores. Consequently, this study represents the first mitochondrial proteome analysis of infected macrophages.

Inhibition of Bacillus cereus Growth and Toxin Production by Bacillus amyloliquefaciens RD7-7 in Fermented Soybean Products

  • Eom, Jeong Seon;Choi, Hye Sun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.44-55
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    • 2016
  • Bacillus cereus is a gram-positive, rod-shaped, spore-forming bacterium that has been isolated from contaminated fermented soybean food products and from the environment. B. cereus produces diarrheal and emetic toxins and has caused many outbreaks of foodborne diseases. In this study, we investigated whether B. amyloliquefaciens RD7-7, isolated from rice doenjang (Korean fermented soybean paste), a traditional Korean fermented soybean food, shows antimicrobial activity against B. cereus and regulates its toxin gene expression. B. amyloliquefaciens RD7-7 exhibited strong antibacterial activity against B. cereus and inhibited the expression of B. cereus toxin-related genes (groEL, nheA, nheC, and entFM). We also found that addition of water extracts of soybean and buckwheat soksungjang (Korean fermented soybean paste made in a short time) fermented with B. amyloliquefaciens RD7-7 significantly reduced the growth and toxin expression of B. cereus. These results indicate that B. amyloliquefaciens RD7-7 could be used to control B. cereus growth and toxin production in the fermented soybean food industry. Our findings also provide a basis for the development of candidate biological control agents against B. cereus to improve the safety of fermented soybean food products.

The Physicochemical Stabilities and Antimicrobial Activities of Pigment Extracts from Zooshikella sp. 17TA (Zooshikella sp. 17TA 색소 추출물의 물리화학적 안정성과 항균활성)

  • Park, Jae-Myeong;Park, Jin-Sook
    • Journal of Marine Bioscience and Biotechnology
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    • v.11 no.2
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    • pp.89-93
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    • 2019
  • In this study, the stability of the extracted natural pigments against light, temperature, pH, metal ions, and antimicrobial activity were evaluated in marine bacteria Zooshikella sp. 17TA. The pigment of the strain used in the study was red with maximum absorption at a wavelength of 541 nm. The stability of the pigment was evaluated by measuring the absorbance while preserving for 15 days and examining the retention rate. After 15 days of irradiation, the pigment of this bacterium showed 98% retention in the dark and 91% retention in the temperature range of -20℃ ~ 30℃. When the pH was in the range 4-7, the retention was about 80%, and the retention rate was higher than 85% for all kinds of metal ions except for CuCl2, ZnCl2, and KCl. The bacterial pigments showed high stability under the given irradiated pH, temperature, and metal ion conditions and had shown activity against gram-positive strains. These results suggest that this highly conserved microbial pigment can be applied to the food industry.

Purification and Properties of Cyclodextrin Glucanotrnsferase Synthesizing $2-O-{\alpha}-D-Glucopyranosyl{\;}_{L}-Ascorbic$ Acid from Paenibacillus sp. JB-13

  • Bae, Kyung-Mi;Kim, Sung-Koo;Kong, In-Soo;Jun, Hong-Ki
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.242-250
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    • 2001
  • A Gram-positive bacterium (strain JB-13) that was isolated from soil as a producer of cyclodextrin glucanotransferase (CGTase) [EC 2.4.1.19] was identified as Panibacillus sp. JB-13. This CGTase could catalyze the transglucosylation reaction from soluble starch to L-ascorbic acid (AA). A main product formed by this enzyme with ${\alpha}-glucosidase$ was identified as $2-O-{\alpha}-D-glucopyranosyl{\;}_{L}-ascorbic$ acid (AA-2G) by the HPLC profile and the elemental analysis. CGTase was purified to homogeneity using ammonium sulfate fractionation, ion-exchange chromatography on DEAE-Seohadex A-50, and gel chromatography on Sephacryl S-200HR. The molecular weight was determined to be 66,000 by both gel chromatography and SDS-PAGE. The isoelectric point of the purified enzyme was 5.3. The optimum pH and temperature was PH 7.0 and $45^{\circ}C$ respectively. The enzyme was stable in the range of pH 6-9 and at temperatures of $75{\circ}C$ or less in the presence of 15 mM ${CaCl_2}.\;{Hg^2+},\;{Mn^+2},{Ag^+},\;and\;{Cu^2+}$ all strongly inhibited the enzyme's activity.

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Biodegradation of Phenanthrene by Psychrotrophic Bacteria from Lake Baikal

  • AHN TAE-SEOK;LEE GEON-HYOUNG;SONG HONG-GYU
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1135-1139
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    • 2005
  • Psychrotrophic phenanthrene-degrading bacteria were identified in the sediment samples collected from Lake Baikal, Russia. Among 70 phenanthrene-degrading isolates, the seven that had the highest phenanthrene-degradation rates were identified by 16S rDNA sequencing. Isolate P25, identified as the Gram-positive rod-shaped organism Rhodococcus erythropolis, had the highest growth and degradation rate at $15^{\circ}C$. It could remove $26.0\%$ of 100 mg $1^{-1}$ phenanthrene in 20 days at $15^{\circ}C$, and degradation was less at $5^{\circ}C\;and\;25^{\circ}C$. The addition of surfactants to enhance degradation was tested. Brij 30 and Triton X-100 inhibited degradation at all surfactant concentrations tested, but Tween 80 stimulated phenanthrene degradation, especially at low concentrations. When $20{\times}$ CMC (critical micelle concentration) of Tween 80 was added, $38.0\%$ of 100 mg $1^{-1}$ phenanthrene was degraded in 12 days at $15^{\circ}C$. This psychrotrophic phenanthrene-degrading bacterium is a candidate for use in bioremediation of polycyclic hydrocarbon contamination in low temperature environments.