• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.706-709
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• 2002

Bacterial bioluminescence has been known to be a highly valuable reporter system for its potential application as an effective and simple environmental monitoring method for toxic compounds. In this short report, we constructed a streptomycetes-Escherichia coli shuttle vector-containing bioluminescence system and evaluated its potential application for toxic compounds monitoring. The luxAB biolurninescence genes from Vibrio harveyi were cloned into a streptornycetes-E. coli shuttle vector (named pESK004) and functionally expressed in Streptomyces lividans. The recombinant S. lividans containing pESK004 exhibited an optimal biolurninescence at the optical density ($OD_{600\;nm}$) of 0.4-0.5 and aldehyde concentration of 0.005%. When the recombinant bioluminescence streptomycetes was exposed to a toxic compound such as heavy metals, chlorinated phenols, or pesticides, the bioluminescence was decreased proportionally to the concentration of toxic compound in the assay mixture. The $EC_{50}$ (effective concentration to decrease 50% of the bioluminescence prior to exposure) values in the recombinant biolurninescence streptomycetes for mercury, 2,4-dichlorophenol, and malathion were measured at 2.2 ppm, 144.0 ppm, and 82.4 ppm, respectively. The degree of sensitivity and specificity pattern toward these toxic compounds characterized in this recombinant bioluminescence streptomycetes were unique when compared with previously reported bacterial bioluminescence systems, and this revealed that a recombinant bioluminescence streptomycetes might provide an alternative or complementary system for potential environmental monitoring.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.94-99
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• 2000

Bioluminescence is being used as a prevailing reporter of gene expression in microorganisms and mammalian cells. Bacterial bioluminescence draws special attention from environmental biotechnologists since it has many advantageous characteristics, such as no requirement of extra substractes, highly sensitive, and on-line measurability. Using bacterial bioluminescence as a reporter of toxicity has replaced the classical toxicity monitoring technology of using fish or daphnia with a cutting-edge technology. Fusion of bacterial stress promoters, which control the transcription of stress genes corresponding to heat-shock, DNA-, or oxidative-damaging stress, to the bacterial lux operon has resulted in the development of novel toxicity biosensors with a short measurement time, enhanced sensitivity, and ease and convenient usage. Therefore, these recombinant bioluminescent bacteria are expected to induce bacterial bioluminescence when the cells are exposed to stressful conditions, including toxic chemicals. We have used these recombinant bioluminescent bacteria in order to develop toxicity biosensors in a continuous, portable, or in-situ measurement from for air, water, and soil environments. All the data obtained from these toxicity biosensors for these environments were found to be repeatable and reproducible, and the minimum detection level of toxicity was found to be ppb (part per billion) levels for specific chemicals.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.349-353
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• 2002

Two recombinant bacteria containing luxAB showed an increased tolerance to stresses associated with lyophilization, when the cells were freeze-dried in the presence of trehalose. In the case of a recombinant, UV2, only $2.5\%$ of the original bioluminescence and $2.7\%$ of the cell viability were restored after 4 h of freeze-drying without trehalose, which implies that the cells were heavily damaged during the dehydration. To improve these losses, trehalose was added before freeze-drying using different modes. Trehalose increased the bioluminescence and the viability of freeze-dried UV2 under all conditions tested, and it was also observed that the addition of trehalose to the cultures (final concentration of 0.08 M) for 15 min before the freeze-drying resulted in the restoration of $45\%$ of the original bioluminescence and $50\%$ of the cell viability. Trehalose also showed a similar efficacy with the other luminescent recombinant, YH9. Therefore, it was tentatively concluded that trehalose played a role as a protective agent in the freeze-drying of bacterial cells.

• KSBB Journal
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• v.14 no.4
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• pp.403-407
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• 1999

Various materials including sodium alginate, k-carragreenan, collagen and polyacrylamide were studied in order to maintain stability of bioluminescence of P. phosphoreum for the purpose of continuos monitoring of toxic subtances. Collagen and polycryamide were shown to be inadequate for immobilization of p. phosphoreum since the bioluminescence decreased when cells were mixed with such materials. In case of k-carrageenan, the bioluminescence was stable when compared with collagen and polyacryamide. However, the k-carrageenan was not suitable for immobilization of p. phosphoreum as cells could not be mixed with the material properly in temperature at which gel formation already occurred. P . phosphoreum must be treated at low temperature below that of gel formation since these are psychrophilic luminescent bacterial. When cells were immobilized on sodium alginate, the bioluminescence was stably maintained for 20 minutes.

• Journal of Ginseng Research
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• v.25 no.3
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• pp.127-129
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• 2001

Bioluminescence technique was applied to ginseng powders. ATP bioluminescence can be used as a rapid method that can be implemented for microbiological monitoring of contaminated ginseng powders. The RLU (relative light units) of ATP was proportion to bacterial CFU (colony forming units) when in high contaminated ginseng powders ($\geq$ about 1.0$\times$10$^4$CFU/g). However, when in low contaminated ginseng powders ($\times$10$^4$CFU/g), the RLU was not proportion to CFU, respectively.

• Korean Journal of Food Science and Technology
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• v.31 no.5
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• pp.1345-1348
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• 1999

The utility of a bioluminescence adenosine triphosphate(ATP) assay method for estimating bacterial levels in mackerel(Scomber japonicus) was investigated. Mackerel was stored at $1^{\circ}C$ throughout 10 days and its RLU(relative light unit) and APC(aerobic plate count) was determined. The ATP bioluminescence assay was validated during the storage of 32 samples, resulting in an agreement between the ATP assay and standard plate count methods of over 90% credibility. Therefore, ATP bioluminescence assay was considered as a rapid and near real-time means in estimating the microbial load on mackerel skin.

• Journal of Food Hygiene and Safety
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• v.15 no.3
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• pp.252-255
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• 2000

This study was conducted to investigate the application of ATP bioluminescence to measure the degree of microbial contamination from raw meat, meat processing and milk processing lines. Samples collected from slaughter house, meat and milk processing plants were tested for estimation of bacterial number by using ATP bioluminescence and conventional method. The former result was transffered to R-mATP value(log RLU/ml), and the latter transffered to CFU(log/ml). Correlation coefficient(r) between aerobic counts(CFU, log/ml) and R-mATP(log RLU/ml) value was 0.93(n=408). R-mATP of aerobic counts from beef, pork, chicken was 0.93(n=220), and that was 0.93(n=187) between meat processing and dairy processing plants. In addition, Correlation coefficient(r) between aerobic counts and R-mATP was 0.87(n=252) under 1$\times$10${^5}$/ml of bacterial count and 0.74(n=152) over 10${^5}$ respectively.

• Proceedings of the Microbiological Society of Korea Conference
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• 1991.04a
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• pp.209-212
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• 1991

• Environmental Engineering Research
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• v.16 no.1
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• pp.41-45
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• 2011

The aim of this investigation was to demonstrate a rapid bioluminescence bioassay for comparison of the toxicity of whole solids and the aqueous extracts of various environmental solid samples. With regard to the toxicities, those for the soil extracts were mostly found to be lower than those of whole soils, which may have been caused by un-extracted pollutants or dilution during the extraction process. Solid samples from dam-reservoir sediments and municipal refuses were also tested. The toxicities of the solid extracts (0-34%; refuses and sediments) were much lower than those of the whole solids (13-91%). The bioluminescence inhibition test indicated that the harmful effects of the contaminated solids samples were greater than those of the solid extracts.

• Journal of Microbiology
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• v.38 no.2
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• pp.74-79
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• 2000

For the demonstration of a novel riboflavin kinase assay method based on the bacterial bioluminescence, partially purified riboflavin kinase was prepared from bovine liver through ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography. Using bacterial luciferase from Photobacterium phosphoreum and the dithionite reduction method, and easy, safe, and fast assay method was established. The optimal temperature, pH, Km values form riboflavin and ATP of boving liver riboflavin kinase determined with this luminescence method were 35$^{\circ}C$, pH 7, 15.3${\mu}$M and 8.3.${\mu}$M, respectively. The detection limit of FMN produced by riboflavin kinase was in the range of 200 pM to 4${\mu}$M which is comparable to the HPLC-fluorescence detection method, while the detection time for each assay was less than 15 sec compared to the HPLC method which requeires at least 10 min for completion.