• Title/Summary/Keyword: Bioluminescence

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Chemical Contamination and Toxicity of Sediments from the Gunsan Coast, Korea

  • Lee, Wan-Seok;Choi, Minkyu;Hwang, Dong-Woon;Lee, In-Seok;Kim, Sook Yang
    • Fisheries and Aquatic Sciences
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    • v.15 no.3
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    • pp.241-250
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    • 2012
  • Polycyclic aromatic hydrocarbons (PAHs), butyltins (BTs), nonylphenol (NP), and fecal sterols concentrations in sediments were investigated from Gunsan coast of Korea to evaluate organic pollution from anthropogenic activities. Sediment toxicity was also examined by bacterial bioluminescence toxicity test (Vibrio fischeri). The concentrations of 16 PAHs in sediments ranged from 67.9 to 425 ng/g dry wt; BTs ranged from 2.79 to 14.1 ng Sn/g dry wt; NP ranged from 20.7 to 2171 ng/g dry wt; and coprostanol, a fecal sterol, ranged from 7.60 to 245 ng/g dry wt. Effective concentration 50% ($EC_{50}$) of sediments ranged from 0.38 to 23.8 mg/mL. Most of the chemicals were present at levels lower than or comparable to the previously reported values from Korea. However, NP levels in the present study were in the high range of levels reported from the Korean coast, and 40% of the measured samples exceeded screening and ecotoxicological values of NP suggested by the Netherlands and Canada. This suggests that an ongoing source of NP is a serious concern in the Gunsan coast. High levels of contaminants were found in the proximity of potential sources, such as the outfall of a wastewater treatment plant for NP, an anthracite-fired power plant for PAHs, and ports for BTs, fecal sterols, and sediment toxicity. This indicates that Gunsan coast has various potential sources of marine sediment contaminants.

Bioassessment and Comparison of Toxicity of Arsenics based on the Results of Various Bioassays (다양한 생물 검정법에 근거한 비소의 위해성 평가 비교)

  • Kong, In-Chul;Kwon, Hyo-Jung;Ko, Kyung-Seok
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.8
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    • pp.795-801
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    • 2010
  • The acute toxicity of arsenic compounds was assessed and compared using following four bioassays; bioluminescence activity of the recombinant strain RB1436, germination of four different seeds, ${\alpha}$-glucosidase activity produced by Bacillus lichemiformis, acute genetic revertant mutation using mutant strain Salmonella typhimurium. Different sensitivities were observed among tested bioassays, but generally the toxicity by arsenite was greater than that of arsenate. Among tested four seeds, sensitivities of Lactucus and Raphanus were greater than others, and these two seed types were appeared as proper type for bioassay. High revertant mutation ratio (5.1) was observed with 1 mg/L arsenite, indicating high mutagenicity. The sensitivity of ${\alpha}$-glucosidase activity on arsenic compounds was much lower than other methods. The evaluation of interactive toxic effects using various bioassays may comprise a useful tool for the bioassessment of environmental pollutants.

Development of a Novel ATP Bioluminescence Assay Based on Engineered Probiotic Saccharomyces boulardii Expressing Firefly Luciferase

  • Ji Sun Park;Young-Woo Kim;Hyungdong Kim;Sun-Ki Kim;Kyeongsoon Park
    • Journal of Microbiology and Biotechnology
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    • v.33 no.11
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    • pp.1506-1512
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    • 2023
  • Quantitative analysis of adenosine triphosphate (ATP) has been widely used as a diagnostic tool in the food and medical industries. Particularly, the pathogenesis of a few diseases including inflammatory bowel disease (IBD) is closely related to high ATP concentrations. A bioluminescent D-luciferin/luciferase system, which includes a luciferase (FLuc) from the firefly Photinus pyralis as a key component, is the most commonly used method for the detection and quantification of ATP. Here, instead of isolating FLuc produced in recombinant Escherichia coli, we aimed to develop a whole-cell biocatalyst system that does not require extraction and purification of FLuc. To this end, the gene coding for FLuc was introduced into the genome of probiotic Saccharomyces boulardii using the CRISPR/Cas9-based genome editing system. The linear relationship (r2 = 0.9561) between ATP levels and bioluminescence generated from the engineered S. boulardii expressing FLuc was observed in vitro. To explore the feasibility of using the engineered S. boulardii expressing FLuc as a whole-cell biosensor to detect inflammation biomarker (i.e., ATP) in the gut, a colitis mouse model was established using dextran sodium sulfate as a colitogenic compound. Our findings demonstrated that the whole-cell biosensor can detect elevated ATP levels during gut inflammation in mice. Therefore, the simple and powerful method developed herein could be applied for non-invasive IBD diagnosis.

Correlation Analysis of Inspection Results and ATP Bioluminescence Assay for Verification of Hygiene Status at 5 Star Hotels in Korea (국내 주요 5성급 호텔의 위생실태 조사와 ATP 결과의 상관분석 평가 연구)

  • Kim, Bo-Ram;Lee, Jung-A;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.36 no.1
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    • pp.42-50
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    • 2021
  • Along with the rapid growth of the food service industry, food safety requirements and hygiene are increasing in importance in restaurants and hotels. Accordingly, there is a need for quick and practical monitoring techniques to determine hygiene status in the field. In this study, we investigated 5 domestic 5-star hotels specifically, personal hygiene (hands of workers), cooking utensils (knife, cutting board, food storage container, slicing machine blade, ice-maker scoop) and other facilities (refrigerator handle, sink). In addition, we examined the hygiene management status of customer contact points (tongs for buffet, etc.) to derive the correlation between the ATP values as a, a verification method. As a result of our five-hotel survey, we found that cooking utensils and personal hygiene were relatively sanitary compared to other inspection items (cookware 92.2%, personal hygiene 91.4%, facilities and equipment 76.19%, customer contact items 88.6%). According to our ATP-based mothod, kitchen utensils (51 ± 45 RLU/25㎠) were relatively clean compared to other with facilities and equipment (167 ± 123 RLU/25㎠). In the present study, we also evaluated the usefulness of the ATP bioluminescence method for monitoring surface hygiene at hotel restaurants. After correlation analysis of surveillance of hygienic status points and ATP assay, most results showed negative and high correlation (-0.64--0.89). Our ATP assay (92 ± 67 RLU/25㎠) of each item after cleaning showed signigicantly reduced results compared to the ATP assay (1020 ± 1254 RLU/25㎠) for normal status, thereby indicating its suitability as a tool to verify the validity of cleaning. By our results, ATP bioluminescence could be used as an effective tool for visual numerical evaluation of invisible contaminants.

The Lux Genes and Riboflavin Genes in Bioluminescent System of Photobacterium leiognathi Are under Common Regulation

  • Sung, Nack-Do;Lee, Chan-Yong
    • Journal of Photoscience
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    • v.11 no.1
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    • pp.41-45
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    • 2004
  • The key riboflavin synthesis genes are located immediately downstream of luxG in the lux operon from Photobacterium leiognathi. It is of interest that a site capable of forming a rho-independent terminator does not appear to be present between luxG and ribE in our previous data. These results raise the question of whether the transcription of lux and rib genes is integrated or not. In order to answer the question, in vivo transcriptional assay and Southern blot were examined. These studies demonstrate that neither transcriptional terminator nor promoter site is present in the intergenic region between of lux and rib genes as well as that the riboflavin genes are single copy in a chromosome of Photobacterium leiognathi.

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Bioluminescent Assay of ${\alpha}$-Oxidase from Cucumis sativus using Bacterial Luciferase-Coupled Reaction

  • Cho, Ki-Woong
    • BMB Reports
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    • v.33 no.4
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    • pp.353-357
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    • 2000
  • A new assay method of ${\alpha}-Oxidase$ (fatty acid : oxygen dioxygenase, 1-decarboxylating) was developed using a bioluminescence reaction system of marine luminous bacterium, Photobacterium phosphoreum. ${\alpha}$-Oxidase was isolated from a cucumber (Cucumis sativus). Pentadecanoic acid was used as a substrate, and the product, tetradecanal, was analyzed with a bacterial luciferase-coupled reaction. Initial light intensity was directly related to the concentration of tetradecanal in the range of 1 nM to 10 ${\mu}M$. Optimal pH and temperature were 7.5 and $25^{\circ}C$, respectively. Optimal pentadecanoic acid concentration in a standard assay of ${\alpha}$-oxidase was 0.1 mM. The Km value of pentedecanoic acid was $85{\mu}M$. This method is straightforward, rapid, convenient, and easy. Its needs no treatment or extraction of reaction mixture.

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Evaluation of Environmental Toxicities for Priority Water Pollutants in a Small Watershed by Bioassays - Comparision between Lettuce Seed Germination Test and Microtox Bioassay - (생물학적 검정법을 이용한 소규모 수계내 수질 오염물질의 환경독성 평가 -상추씨 발아시험과 Microtox 시험 비교-)

  • 이지나;황인영
    • Environmental Analysis Health and Toxicology
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    • v.14 no.4
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    • pp.135-144
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    • 1999
  • Environmental toxicities of priority water pollutants were evaluated by two selected bioassays, Lettuce seed germination/elongation test and Microtox acute toxicity test. Toxic chemicals (heavy metals, polycyclic aromatic hydrocarbons, and phenolic compounds) inhibited the germination rate and root elongation of Lettuce seed, as well as the bioluminescence of Microtox bacteria. When test biota were exposed to target chemicals, the sensitivity of Lettuce bioassay was relatively lower than that of Microtox bioassay. However, Lettuce bioassay may be a good candidate for prescreening the environmental toxicities of priority water pollutants, since the testing method with Lettuce seed was relatively easier and more economic than with Microtox bacteria. Toxicity tests were conducted to compare the validity and sensitivity of both bioassays for sediment from a small stream passed through urban area as well as leachate from a municipal solid waste landfill. From experimental results, we found that Lettuce test and Microtox test are compensated each other as a battery of bioassay for evaluating the environmental toxicities of field samples obtained from a small stream contaminated by pollutants.

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Construction of Bioluminescent Escherichia coli from lux Operon and Heat Shock Promoter for the Detection of Toxic Substances (lux Operon과 Heat Shock Promoter 유전자 재조합을 통한 독성물질 탐지용 대장균의 개발)

  • 유승오;이은관;김현숙;정계훈;전억한
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.278-285
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    • 1999
  • In order to use heat shock promoter for the detection of toxic substances, dnaK promoter was amplified from E. coli genomic DNA by using a polymerase chain reaction(PCR) followed by sequencing and sub-cloning into the multi-cloning site of the plasmid, pUCD615. The pUCD615 is a broad-host-range vector containing promoterless lux operon originated from V.fischeri. The recombinant plasmid was transfered to E. coli DH5$\alpha$ through electroporation. The recombinant E. coli showed several patterns of bioluminescent responses to ethanol stress. The bioluminescent E. coli also showed responses to other toxic substances including FeK3(CN)6, CdCl2, p-nitrophenol and HgCl2. The increases of RLU(Relative Light Unit) were observed at 100ppm of FeK3(CN)6, 10ppm and 100ppm and 100ppm of CdCl2, 1ppm of 10ppm of p-nitrophenol and at 1ppm of HgCl2.

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