• KSBB Journal
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• 제14권1호
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• pp.91-95
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• 1999

P. phosphoreum의 고정화에 있어 중요한 것은 Matrix의 선택이며, Matrix로서 sodium algnate만을 사용하여 고정화하는 것보다는 strontnum chloride를 첨가하였을 때 세포의 bio-iummescence 유지도가 증가하였다. 저장 온도에 따른 bio-iummescence 유지도와 관련하여 -$70^{\circ}C,\;-20^{\circ}C,\;20^{\circ}C$에서 저장한 세포의 경우 저장 1일 후에 급격한 bioluminescence의 감소를 보였으나 $4^{\circ}C$ 에서 저장한 세포는 bioluminescence의 유지도가 15일 이상 이어졌다. 따라서 P. phosphoreum의 bioluminescence 안정성에 있어 가장 좋은 결과를 나타낸 것은 2.5%(W/W)sodium alginate와 0.3M(W/V)strontnum chloride 를 사용하여 고정화한 세포였으며 저장 온도는 $4^{\circ}C$였다.

• Journal of Microbiology and Biotechnology
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• 제7권4호
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• pp.242-249
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• 1997

Stability of bioluminescence was investigated with Photobacterium phosphoreum immobilized on the strontium alginate in order to develope continuous real time monitoring of pollutants. The stability of bioluminescence emission was improved by prolonged aging time. The aging time of ${\geq}40$ min and the cell concentration of ${\leq}0.6\;of\;OD_660$ were selected for the immobilization of P. phosphoreum to give linearity between cell concentrations and bioluminescence intensity. In sensitivity tests using phenol, it was found that this compound quenched bioluminescence proportional to the concentration without lowering of cell growth. The lower value for maximum quenching ($q_s$) and higher dissociation constant ($K_s$) were observed with strontium-alginate immobilized cells compared to free cells. The response of bioluminescence to toxicants was evaluated with the immobilized luminescent bacteria. The sensitivity of the immobilized cells was found to be good in response to toxicants, 4-nitrophenol, salicylate and cadmium, when evaluated with a specific rate of bioluminescence quenching.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2000년도 Proceedings of 2000 KSAM International Symposium and Spring Meeting
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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.

• 대한환경공학회지
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• 제30권1호
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• pp.79-84
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• 2008

본 연구에서는 유전자 재조합 발광균주, Pseudomonas putida mt-2 KG1206,의 발광 촉진조건을 석유계 탄화수소 오염 지하수에 적용 가능성에 대해 조사하였다. 일반적으로 순수 유도제 오염원에 대한 발광 강도는 m-methyl benzyl alchohol > m-toluate > toluene > m-xylene > benzoate > p-xylene > o-xylene의 순으로 나타났다. 일반적으로 지하수에 오염된 유도제 오염원의 농도에 따라 발광 정도가 관찰되었다. 그러나 일부 지하수 시료의 경우에는 낮은 농도에 높은 발광과 같은 현상을 관찰할 수 있었다. 이러한 현상은 환경시료의 수질 복잡성에 따른 영향일 것이다. 선행연구에서 발광촉진제로 알려진 SL (sodium lactate)와 KNO$_3$에 의한 발광영향을 비교하였다. 모두 발광을 촉진하였지만, KNO$_3$가 조금 더 촉진력이 강한 것으로 조사되었다. 질소화합물(20 g KNO$_3$/L) 첨가군 시료의 발광균주는 첨가하지 않은 대조군에 비해 뚜렷하게 발광이 높게 나타났다. 따라서 일반적으로 낮은 유도제 오염원 함유 시료에 대해서는 발광촉진 조건이 지하수 오염원 검출 및 모니터링을 용이하게 할 수 있을 것이다. 또한 비슷한 특성을 가진 다양한 발광균주를 더욱 효율적으로 오염지역의 기초평가에 이용할 수 있는 조건이 될 수 있을 것이다.

• Journal of Microbiology and Biotechnology
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• 제12권2호
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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.

• Journal of Microbiology and Biotechnology
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• 제7권4호
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• pp.250-257
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• 1997

A new sensing system based on the immobilization of luminescent bacteria, photobacterium phosphoreum, was proposed for continuous real-time monitoring of pollutants. The response curves demonstrate that Photobacterium phosphoreum immobilized on the strontium alginate were very sensitive to seven reference chemicals used. The significant inhibitory concentrations for bioluminescence emission were 5 ppm for Pb$(NO_3)_2$), $NiCl_2$, $CdCl_2$, 50 ppm for $NaASO_2$, 0.1 ppm for $HgCl_2$, 0.5 ppm for pentachlorophenol and less than 5 ppm for SDS, respectively. The alginate mixed-cells (AMC) retained their luminescence during experimental period (29 days) under storage condition of $-80^{\circ}C$. The variables affecting performance of continuous flow through monitoring (CFTM) was optimized in order to ensure stability and efficiency. The flow through cell with strontium-alginate immobilized luminescent bacteria was tested with salicylate and 4-nitrophenol. A rapid response of luminescence was recorded by time drive mode in bioluminescence spectrometer after exposure to both toxicants.

• 한국지하수토양환경학회지:지하수토양환경
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• 제18권4호
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• pp.32-39
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• 2013

This research focused on the effects of heavy metals on the biomonitoring activity of genetically engineered bioluminescent bacteria, Pseudomonas putida mt-2 KG1206. KG1206 was exposed to single or binary mixtures of different heavy metals as well as soils contaminated with heavy metals. In case of single exposure with different inducer pollutant, the toxicity order was as followings : As(III) > Cd, As(V) >> Cu, Cr(VI). The toxic effects of the binary mixtures was compared to the expected effect based on a simple theory of probabilities. The interactive effects were mostly additive, while in few cases antgonistic and synergistic mode of action was observed for some concentration combinations. No considerable correlation was found between the total metal contents in soils and the bioluminescence activity of each sample. However, statistically significant differences (p = 0.0102) were observed between two groups, classified based on arsenite contamination. These results demonstrate the usage of recombinant bioluminescent microorganism in biomonitoring and the complex interactive effects of metals.

• 한국식품과학회지
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• 제18권2호
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• pp.88-92
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• 1986

Bioluminescence방법에 의하여 세균내에 존재하고 있는 ATP를 추출하여 그양을 정량분석하므로서 제육표면에 생존하고 있는 세균수를 신속하게 측정하는데 응용될 수 있는 가능성을 조사하기 위하여 본 실험을 실시하였으며, 그 결과는 1. Luciferin-luciferase에 작용하는 ATP농도가 $10^{-10}M$부터 $10^{-6}M$까지 증가함에 따라 발생된 빛의 양도 비례적으로 증가했다. 2 . 계육에서 분리동정된 부패균인 E. coli, H, alvei, P. putida, A. hydrophila에서 추출된 ATP 농도와 세균수와의 관계는 거의 일직선으로 나타난다. 3. E. coli에 있는 ATP양은 생존중 거의 변화하지 않는 것으로 나타났다. 4 계육의 저장기간동안 증식된 세균수에 따라 ATP 농도도 비례 적으로 증가되며 이때 상판계수(r)는 0.95로 나타났다. 5. 계육에 오염된 세균수와 ATP 농도와의 관계에서 볼 때 ATP농도가 $10^{-7}M$부근에서부터 부패가 시작된다고 사료된다. 6. 개발된 방법에 의하여 계육표면에 존재하는 세균의 ATP 농도를 측정하여 세균수를 간접적으로 신속히 ( 1시간내) 조사할 수 있다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.212-215
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• 2000

토양오염의 독성을 탐지하기 위해 재조합 발광 박테리아의 고정화를 이용하여 바이오 센서를 제작하였으며 이를 이용하여 대표적 토양오염물질인 PAHs의 독성을 측정할 수 있었다. 또한, 이 바이오 센서를 이용하여 토양오염처리 전후의 처리 효율을 신속히 탐지할 수 있음을 확인하였다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.390-393
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• 2000

재조합 발광미생물인 UV2와 YH9을 보존제로서 trehalose를 혼합한 현탁액을 바로 동결건조한 것과 이 현탁액을 $30^{\circ}C$ incubator에서 15분 동안 진탕배양한 후 동결건조한 것과 trehalose가 첨가된 배지에 약 $6{\sim}7$시간 정도 배양한 배양액을 동결건조한 결과, trehalose를 첨가한 후 15분 동안 진탕배양했을 경우가 UV2는 0.08M의 이상에서 약 45.0%, 50.0%, YH9은 0.16M에서 48.7%, 54.1%로 회복율을 가장 높이는 최적의 조건으로 조사되었다.