• The Korean Journal of Mycology
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• v.48 no.4
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• pp.381-388
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• 2020

Bioluminescence refers to the production and emission of light in living organisms. This phenomenon arises from luciferase-catalyzed oxidation reaction of luciferin. Bioluminescence is widely observed in marine vertebrates and invertebrates, as well as in some microorganisms and fungi. To date, approximately 80 species of fungi have been reported to be luminous. One such example is Omphalotus japonicus, which is a luminous fungus found in Korea. In this study, we examined the bioluminescence of Omphalotus japonicus mycelia. Light emission was detected at the edges of mycelia grown on solid agar medium. Notably, the intensity of bioluminescence was found to be significantly enhanced following wound induction. The increase in light intensity peaked at 3 h after mechanical damage. We also investigated the effects of extreme temperatures on bioluminescence. Unlike mechanical damage, high and low temperatures repressed the light emission from mycelia. Further investigations are required to reveal the physiological and ecological properties of fungal bioluminescent responses to environmental stresses.

• Journal of Korean Society of Environmental Engineers
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• v.31 no.2
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• pp.147-152
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• 2009

In this study, the applicability of alginate-immobilized Pseudomonas putida mt-2 KG1206 on the environments, contaminated with toluene analogs was conducted. Genetically engineered strain KG1206 produces light by direct (m-toluate, benzoate) and indirect (toluene, xylenes) inducers. The protocol for the alginate-immobilization was determined in terms of the cell to alginate ratio, solution, proper number of alginate beads, and other conditions. Maximum bioluminescences of five chemicals by immobilized strain were generally observed in following orders: m-toluate > p-xylene > toluene > o-xylene > m-xylene. In relationship between bioluminescence activity and inducer reduction, initial m-toluate (5 mM) in solution was removed approximately 48% of initial at 5 h exposure, showing continuous decrease of inducer chemical in solution. These results of study with alginate-immobilized beads would be useful, especially, for biomonitoring of contaminated environments with specific compounds, such as petroleum hydrocarbon compounds including toluene analogs.

• Journal of Korean Society of Environmental Engineers
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• v.34 no.2
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• pp.136-142
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• 2012

Various factors affecting on the bioassay were investigated. Experiments with a low mixture ratio (cell to toxicant solution) of 0.5 : 9.5 (v/v) produced observable bioluminescence intensity for assay. Both sodium lactate and potassium nitrate stimulate bioluminescence activity; 2.6~4.0 times of control. Distilled water and MSM, which gave non significant effects on the bioluminescence activity, were determined as proper diluent or extract solutions. A wide range of toxic responses of metals and organics were observed. In general, organics were much less sensitive than metals. Samples collected from eleven sites showed the bioluminescence activity ranging from 29 to 111% of the control. Significant correlation between toxicity and total metal contents was not observed, but the toxicity of two groups, sorted based on the contaminated arsenic concentration in soils, was 44% and 20%, showing considerable differences.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.5
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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.

• Environmental Analysis Health and Toxicology
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• v.19 no.3
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• pp.295-305
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• 2004

본 연구에서는 유전자 재조합 발광 박테리아를 이용하여 농약에 대한 박테리아의 스트레스 반응과 세포 독성을 분석하였다. 15종류의 농약에 대하여 유전자 손상, 생물막 손상, 산화적 손상 및 단백질 손상을 측정할 수 있는 발광 박테리아와 독성 유무로 인한 세포 독성을 측정할 수 있는 발광 박테리아, 5종을 이용하여 스트레스 반응을 분류하고 세포 독성 정토를 분석하였다. 그 결과, 농약의 화학적 구조가 박테리아의 스트레스 반응에 영향을 미치며, 산화과정이 진행 됨에 따라 독성의 작용 기작이 변하는 것을 확인 할 수 있었다. 이와 같은, 유전자 재조합 발광 박테리아를 이용한 생물체내의 독성 메커니즘에 대한 분석은 생태계 유해물질들에 의한 독성을 분석하고 예상하기 위해 적용될 수 있을 것이다.

• Analytical Science and Technology
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• v.23 no.1
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• pp.60-67
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• 2010

A sensitive competitive heterogeneous bioluminescence immunoassay for serotonin was developed using photoprotein, aequorin as a label for the first time with the optimal assay conditions; especially, serotoninavidin conjugate was prepared by Mannich reaction and the synthetic process of serotonin-avidin conjugate was optimized by controlling the initial molar ratios of serotonin, formaldehyde and avidin (1:12,000:25). The developed bioluminescence immunoassay for serotonin showed good sensitivity (LOD of 0.68 ng/mL) with wide area of dynamic range ($5.0{\times}10^{-10}\;M\sim5.0{\times}10^{-7}\;M$). (cf. the range for serotonin in human blood serum is $151{\pm}45\;ng$/mL). In addition, cross-reactivity studies demonstrated that 5-methoxytryptamine showed some cross-reactivity (28.0%), whereas 3-methylindole, melatonin and 5-hydroxylindole-3-acetic acid showed no crossreactivity, and good recoveries were obtained in serum. Thus, this developed method provides a good tool to monitor serotonin in serum.

• Proceedings of the Korea Society of Environmental Biology Conference
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• 2004.12a
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• pp.29-34
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• 2004

• Journal of Korean Society of Environmental Engineers
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• v.30 no.1
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• pp.79-84
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• 2008

This paper describes the application of bioluminescence stimulating agents on a genetically engineered microorganism, Pseudomonas putida mt-2 KG1206, to monitor toluene analogs using in groundwater samples from petroleum hydrocarbon contaminated sites. The maximum bioluminescent response with pure chemicals followed in the order: m-methyl benzyl alchohol > m-toluate > toluene > m-xylene > benzoate > p-xylene > o-xylene. Generally, the bioluminescence production of strain mixed with groundwater samples was dependent on the contaminated total inducer concentrations. However, few samples showed opposite results, where these phenomena may be caused by the complexicity of environmental samples. Two chemicals, SL(sodium lactate) and KNO$_3$, were tested to determine a better bioluminescence stimulant. Both chemicals stimulate the bioluminescence activity of strain KG1206, however, a slightly high bioluminescence was observed with nitrogen chemical. This selected stimulant was then tested on samples collected from contaminated groundwater samples. The bioluminescence activity of all samples mixed with the strain was stimulated with KNO$_3$ amendment. This suggests that the low bioluminescence activity exhibited by the environmental groundwater samples can be stimulated by amending the culture with a proper agent, such as nitrogen compound. These findings would be useful, especially, when strain was used to monitor the groundwater samples contaminated with low inducer contaminants. Overall, the results of this study found the ability of bioluminescence producing bacteria to biosensor a specific group of environmental contaminants, and suggest the potential for more efficient preliminary application of this engineered strain in a field-ready bioassay.

• KSBB Journal
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• v.17 no.6
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• pp.571-576
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• 2002

Recombinant E. coli DH5 ${\alpha}$/pSB311 was made by cloning the genes encoding bacterial luciferase and aldehyde substrate proteins from Photohabdus luminescense, to complement defects of Lumistox, which is normally used in bioassays to monitor toxic substances in water environmental systems. The conditions for stable light production by the recombinant strains were investigated with respect to cell growth stage, cell number, and buffer conditions. The optimum growth stage was a middle-exponential stage with an OD$_{660nm}$ value of 0.6-0.7. ADout 10$^{6}$-10$^{7}$ cells per test tube was optimum for stable light emission. The effect of buffer was not significant if an optimum viable cell number was maintained. The bioluminescence of the recombinant E. coli harboring the lux operon of Photohabdus luminescense was not affected by temperature, while the bioluminescence of Lumistox was temperature sensitive. The recombinant E. coli was more sensitive to heavy metals (Cd, Cu, Hg, Zn) than Lumistox, because it does not require high concentrations of NaCl in the buffer.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.117-120
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• 2000

For detecting toxicity of endocrine disruptors (EDs), rapid, sensitive, and simple methods are needed. Therefore, in this study, a new method in which the different toxic effect of EDs can be monitored using 4 different recombinant bacteria was designed and evaluated. It was found that the recombinant bacteria could monitor the toxic effect, not estrogenic effect, due to EDCs through the measurement of bioluminescence and cell growth rate, which were shown to depend upon a form of cellular toxicity, such as DNA damage, protein damage, oxidative damage, and membrane damage. In addition, it was found that the damage done by EDCs can be divided into several groups based upon the toxic mechanisms of the EDCs