• 제목/요약/키워드: bioluminescent bacteria

검색결과 47건 처리시간 0.033초

Detection of m-toluate in Soils using Bioluminescence Producing Recombinant Bacteria (유전자 재조합 발광균주를 이용한 토양 오염원 m-toluate 탐지)

  • Kong, In-Chul;Kim, Myung-Hee;Jung, Yun-Ho;Ko, Kyung-Seok;Kim, Jae-Gon;Shin, Sung-Chun
    • Journal of Korean Society of Environmental Engineers
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    • 제27권5호
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    • pp.507-512
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    • 2005
  • This research focuses on the development and application of a method for the detection of m-toluate in soils using a genetically engineered bioluminescent bacteria, Pseudomonas putida mt-2 KG1206. KG1206 produces light by direct (m-toluate and benzoate) and indirect (toluene analogs) inducers. For detection of m-toluate in soil system, 9.9 mL strain was amended with 0.1 mL soil ethanol extractant. A high correlation ($r^2>0.97$) was observed between bioluminescence and m-toluate concentration. The unknown concentrations of m-toluate in soil samples were pre-determined using a method developed based on bioluminescence activity of strain with extracted inducers. Values between by LC analysis and bioluminescence activity show moderate statistical results. These results demonstrate the feasibility of recombinant bioluminescent microorganism, engineered to generate a quantifiable bioluminescence signal in response to specific pollutants, may serve as combined sensing and reporting tools in environmental monitoring.

Probing of Potential Luminous Bacteria in Bay of Bengal and Its Enzyme Characterization

  • Balan, Senthil S.;Raffi, S.M.;Jayalakshmi, S.
    • Journal of Microbiology and Biotechnology
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    • 제23권6호
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    • pp.811-817
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    • 2013
  • The present study dealt with the isolation, identification and enzyme characterization of potential luminous bacteria from water, sediment, squid, and cuttle fish samples of the Karaikal coast, Bay of Bengal, India during the study period September 2007 - August 2008. Bioluminescent strains were screened in SWC agar and identified using biochemical tests. As Shewanella henadai was found to be the most common and abundant species with maximum light emission [69,702,240 photons per second (pps)], the optimum ranges of various physicochemical parameters that enhance the luciferase activity in Shewanella hanedai were worked out. The maximum luciferase activity was observed at the temperature of $25^{\circ}C$ (69,674,387 pps), pH of 8.0 (70,523,671 pps), salinity of 20 ppt (71,674,387 pps), incubation period of 16 h (69,895,714 pps), 4% peptone (70,895,152 pps) as nitrogen source, 0.9% glycerol (71,625,196 pps), and the ionic supplements of 0.3% $CaCO_3$ (73,991,591 pps), 0.3% $K_2HPO_4$ (73,919,915 pps), and 0.2% $MgSO_4$ (72,161,155 pps). Shewanella hanedai was cultured at optimum ranges for luciferase enzyme characterization. From the centrifuged supernatant, the proteins were precipitated with 60% ammonium sulfate, dialyzed, and purified using anion-exchange chromatography, and then luciferase was eluted with 500 mM phosphate of pH 7.0. The purified luciferase enzyme was subjected to SDS-PAGE and the molecular mass was determined as 78 kDa.

Coregulation of lux Genes and Riboflavin Genes in Bioluminescent Bacteria of Photobacterium phosphoreum

  • Sung, Nack-Do;Lee, ChanYong
    • Journal of Microbiology
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    • 제42권3호
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    • pp.194-199
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    • 2004
  • Investigation of the expression of the riboflavin (rib) genes, which are found immediately downstream of luxG in the lux operon in Photobacterium phosphoreum, provides more information relevant to the evolution of bioluminescence, as well as to the regulation of supply of flavin substrate for bacterial bioluminescence reactions. In order to answer the question of whether or not the transcriptions of lux and rib genes are integrated, a transcriptional termination assay was performed with P. phoxphoreum DNA, containing the possible stem-loop structures, located in the intergenic region of luxF and luxE ($\Omega$$\_$A/), of luxG and ribE ($\Omega$$\_$B/), and downstream of ribA ($\Omega$$\_$c/). The expression of the CAT (Chloram-phenicol Acetyl Transferase) reporter gene was remarkably decreased upon the insertion of the stem-loop structure ($\Omega$$\_$c/) into the strong lux promoter and the reporter gene. However, the insertion of the structure ($\Omega$$\_$B/) into the intergenic region of the lux and the rib genes caused no significant change in expression from the CAT gene. In addition, the single stranded DNA in the same region was protected by the P. phosphoreum mRNA from the Sl nuclease protection assay. These results suggest that lux genes and rib genes are part of the same operon in P. phosphoreum.

Sediment Toxicity of Industrialized Coastal Areas of Korea Using Bioluminescent Marine Bacteria

  • Choi, Min-Kyu;Kim, Seong-Gil;Yoon, Sang-Pil;Jung, Rae-Hong;Moon, Hyo-Bang;Yu, Jun;Choi, Hee-Gu
    • Fisheries and Aquatic Sciences
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    • 제13권3호
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    • pp.244-253
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    • 2010
  • The quality of marine sediments from the industrialized coastal areas of Korea (Ulsan Bay, Masan Bay, and artificial Lake Shihwa) was investigated using a bacterial bioluminescence toxicity test. Sediment toxicity results were compared with the levels of chemical contamination (trace metals, organic wastewater markers, acid volatile sulfides, total organic carbon). Effective concentration 50% (EC50) of sediments ranged from 0.014 to 1.126 mg/mL, which is comparable to or lower than values in contaminated lakes, rivers, and marine sediments of other countries. Sediment reference index (SRI) ranged from 13 to 1044, based on the EC50 of the negative control sample. Mean average SRI values in Masan Bay and Lake Shihwa were approximately 8 and 9 times as high as that in Ulsan Bay, indicating higher sediment toxicity and greater contamination in the two former regions. Sediment toxicity were strongly associated with the concentrations of some chemicals, suggesting that this test may be useful for determining potential chemical contamination in sediments.

Physiological and Molecular Characterization of a Newly Identified Entomopathogenic Bacteria, Photorhabdus temperata M1021

  • Jang, Eun-Kyung;Ullah, Ihsan;Lim, Jong-Hui;Lee, In-Jung;Kim, Jong-Guk;Shin, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • 제22권12호
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    • pp.1605-1612
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    • 2012
  • The present study concerned the identification and characterization of a novel bacterial strain isolated from entomopathogenic nematodes collected from different regions in Korea. The bacterial isolate M1021 was Gramnegative, bioluminescent, and produced red colonies on MacConkey agar medium. A rod-shaped structure was confirmed by the electron micrograph. Fatty acid composition was analyzed by using the Sherlock MIDI system. The identification was further supported by 16S rDNA sequence analysis, which revealed 96-99% sequence homology with strains of Photorhabdus temperata. The location of the isolated strain of P. temperata in the phylogenetic tree was confirmed and it was named P. temperata M1021. P. temperata M1021 exhibited catalase, protease, and lipase activities when grown on appropriate media supplemented with respective substrates. The culture of P. temperata M1021 exhibited insecticidal activity against the larvae of Galleria mellonella and the activity was the highest after 3-4 days of cultivation with agitating at $28^{\circ}C$ under 220 rpm. Antibacterial activity was also observed against Salmonella Typhimurium KCTC 1926 and Micrococcus luteus KACC 10488.

MOLECULAR NUCLEAR IMAGING FOR TARGETING AND TRAFFICKING

  • Bom Hee-Seung;Min Jung-Jun;Jeong Hwan-Jeong
    • Nuclear Engineering and Technology
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    • 제38권5호
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    • pp.399-404
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    • 2006
  • Noninvasive molecular targeting in living subjects is highly demanded for better understanding of such diverse topics as the efficient delivery of drugs, genes, or radionuclides for the diagnosis or treatment of diseases. Progress in molecular biology, genetic engineering and polymer chemistry provides various tools to target molecules and cells in vivo. We used chitosan as a polymer, and $^{99m}Tc$ as a radionuclide. We developed $^{99m}Tc-galactosylated$ chitosan to target asialoglycoprotein receptors for nuclear imaging. We also developed $^{99m}Tc-HYNIC-chitosan-transferrin$ to target inflammatory cells, which was more effective than $^{67}Ga-citrate$ for imaging inflammatory lesions. For an effective delivery of molecules, a longer circulation time is needed. We found that around 10% PEGylation was most effective to prolong the circulation time of liposomes for nuclear imaging of $^{99m}Tc-HMPAO-labeled$ liposomes in rats. Using various characteristics of molecules, we can deliver drugs into targets more effectively. We found that $^{99m}Tc-labeled$ biodegradable pullulan-derivatives are retained in tumor tissue in response to extracellular ion-strength. For the trafficking of various cells or bacteria in an intact animal, we used optical imaging techniques or radiolabeled cells. We monitored tumor-targeting bacteria by bioluminescent imaging techniques, dentritic cells by radiolabeling and neuronal stem cells by sodium-iodide symporter reporter gene imaging. In summary, we introduced recent achievements of molecular nuclear imaging technologies in targeting receptors for hepatocyte or inflammatory cells and in trafficking bacterial, immune and stem cells using molecular nuclear imaging techniques.

Phytoplankton as Standard Test Species for Marine Ecotoxicological Evaluation (해양생태독성평가를 위한 표준시험생물로서의 식물플랑크톤에 관한 연구)

  • Park Gyung-Soo;Lee Sang-Hee;Lee Seung-Min
    • Journal of Environmental Science International
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    • 제14권12호
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    • pp.1129-1139
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    • 2005
  • A series of experiments was conducted to identify the potential of five phytoplankton species as standard test species for marine ecotoxicological tests. The candidate phytoplankton species are Skeletonema costatum, Heterosigma akashiwo, Prorocentrum micans, Isochrysis galbana, and Tetraselmis suecica. Salinity tolerance and sensitivity on potassium dichromate as a reference material were identified. Toxicity of eleven ocean dumped sewage sludges and four red tide expellent extracts were estimated by the inhibition of population growth rates (PGR) of marine diatom S. costatum, While most species revealed relatively weak tolerance on salinity, T. suecica demonstrated the highest salinity tolerance ranged from $5\~35$ psu and the others $15\~35$ psu. H. akashiwo revealed the highest sensitivity as 72h $IC_{50}$=0.76mg/L and T. suecica the lowest as 72h $IC_{50}$=8.89mg/L on potassium dichromate. Sludge extracts from industrial waste, domestic sewage and livestock farm waste sludge showed high toxicity as 72h $IC_{50}$<$2\%$ and lowest toxicity from filtration bed sludge as 72h $IC_{50}$=$30.50\%$ NOEC (No Observed Effective Concentration) of sludge extract ranged from <$0.4\%$ to $1.6\%$ and this indicated high phytotoxicity of ocean dumped sewage sludge. The test sensitivity of phytoplankton PGR inhibition was much higher than those of marine rotifer Brachionus plicatilis mortality test and bioluminescent inhibition test by marine bacteria Vibrio fischeri, and comparable with the sea urchin (Strongylocentrotus intermedius) fertilization test. As a result the phytotoxicity test using phytoplankton PGR inhibition ($IC_{50}$) must be a useful tool for marine phyto-toxicological evaluation of ocean dumped materials.

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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    • 제16권11호
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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.

Expression of the Genes Involved in the Synthesis of Riboflavin from Photobacterium species of Bioluminescent Marine Bacteria (해양 발광 박테리아 Photobacterium Species의 Riboflavin 생합성에 관여하는 유전자들의 발현)

  • 이찬용
    • Korean Journal of Microbiology
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    • 제36권1호
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    • pp.1-7
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    • 2000
  • The genes involved in riboflavin synthesis (ribI, II, III, and IV) were found immediately downstream of luxG in the lux operon from Photobacterium species. The single stranded DNA containing the intergenic region of lux genes and rib genes from Photobacterium phosphoreum was fully protected by P. phosphoreum mRNA from the S1 nuclease mapping assay suggesting that a transcriptional terminator was not present in the region. In addition, the levels of riboflavin synthase activity in P. phosphoreum was increased during the development of bacterial bioluminescence in the same fashion as the luciferase and fatty acid reductase activities. Insertion of the Photobacterium leiognathi DNA extending from luxB to ribII, between a strong lux promoter and a reporter gene (chloramphenicol acetyltransferase, CAT) and transferred by conjugation into P. leiognathi, did not affect expression of reporter gene. Moreover the CAT gene was not expressed in an analogous construct missing the lux promoter indicating that a promoter was not present in this region. Based on the data here, it can be concluded that the lux genes and rib genes in Photobacterium species are under common regulation.

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An Adaptedness Assessment for Beach Nourishment Utilization of Dredged Materials in Hwasun-Port (화순항내 준설토사의 양빈재 유효활용을 위한 적합성 평가)

  • Choi, Jin-Hwan;Lee, Sang-Houck
    • Journal of the Korea Academia-Industrial cooperation Society
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    • 제16권4호
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    • pp.2940-2944
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    • 2015
  • Dredged materials for effective use in Hwasun-port pollution analysis, Ni(nickel) in excess of the standard showed that the whole point. However, according to the revised standards effective utilization exceeds the reference value, even if no anthropogenic sources Pearl ratio less than 10% of the average weight of the ecotoxicological tests have passed, and if so it can be used to beach nourishment. Therefore, the average weight ratio of pearl and ecotoxicological tests were conducted by marine environment official test method. Mud to 5.37% of the average weight ratio lower than the reference value of 10% that was investigated. Ecotoxicity test using bioluminescent bacteria relative luminescence inhibition in the 0 to 9.13% with no toxicity was determined using benthic amphipods in the control and the test sample relative to the average survival rate is 0% with no toxicity was determined. Therefore, the revised effective use meets all the criteria that can be used to beach nourishment, so that the suitability was assessed.