고정화 Photobacterium phosphoreum과 휘발성 독성 물질과의 반응성

이은수,전억한 경희대학교 생명자원과학연구원 1999 硏究論文集 Vol.20 No.-

휘발성 독성 물질 측정용 biosensor로서 P phosphoreum을 sodium alginate로 고정화하여 독성물질에 대한 반응성을 조사한 결과 각각의 독성물질마다 특이한 hnearity 관계식이 설정되는 농도 범위가 있었고 이를 이용하여 EC_50을 계산할 수 있었다 IC_50과 비교해보면 EC_50을 이용하는 것이 시간과 비용을 줄일 수 있고 낮은 농도의 독성 물질에 대해 더 민감한 반응을 나타내었다 특히, 휘발성이 강한 독성 물질 일수록 휘발성 방지를 위해서는 세포를 고정화하는 것이 더 효과적이었다 따라서 휘발성 독성물질에 대해서 free cell보다는 고정화한 세포가 더 민감하게 반응하였고 sodium alginate 고정화 세포보다는 strontium alginate로 고정화된 세포가 민감한 반응을 나타내었다 Immobilization method was investigated in order to maintain the stability of bioluminescence of Photobacterium for monitoring the volatile toxic substances. Sodium alginate was used as an immobilization matrix for P. phosphoreum. For the relationship between bioluminescene of immobilized P. phosphoreum and toxic substances, kinetic parameters of specific rate (μ), ratio of bioluminescence intensity (Io/I), and gamma (γ) value were estimated. The bioluminescence intensity was found proportional to the concentration of toxic substances and the free cells showed more sensitivity than immobilized cells when volatile substances were exposed to the cells. However, bioluminescence increased slightly with time because of volatility. Furthermore, P. phosphoreum immobilized on strontium alginate were rather favored in comparison with cells immobilized on sodium alginate for the response to substances used.

Inulinase와 투과성이 향상된 Zymomonas mobilis를 이용한 Jerusalem artichoke로 부터의 sorbitol 생산

김인철,전억한 경희대학교 부설 식량자원개발연구소 1991 硏究論文集 Vol.12 No.-

The use of Jerusalem artichoke containing p-1, 2-fructose oligomer in the production of sorbitol that is used as food additive and converted to L-sorbose have been studied. Coimmobilization method of inulinase and oxidoreductase was considered for the simultaneous hydrolyzation of inulin and conversion of glucose and fructose liberated from inulin to sorbitol. Both inulinase and oxidoreductase were immobilized in chitin(5%, w/v) and k-carrageenan(4%, w/v). The activity of oxidoreductase was specified by permeabilization of Zymomonas mobilis cell with 0.2% CTAB (cetyltrimethylammoniumbromide). The use of inulinase for hydrolyzation of inulin resulted in the achievement of each 36.65g/ℓ of glucose and 85.32g/ℓ of fructose which are valuable substrates for sorbitol production. Using these hydrolyzates, accumulation of 36.13g/ℓ of sorbitol occurred at 38 C and pH6.2 in the reactor. When permeabilized cell and inulinase were coimmobilized, sorbitol produced at 28.lg/ℓ. That is low compared with 36.13g/ℓ in separated reactor system.

Photobacterium phosphoreum을 고정화하기 위한 최적 Carboxymethylcellulose(CMC) 담체의 농도결정

이용제,전억한 경희대학교 생명자원과학연구원 1999 硏究論文集 Vol.20 No.-

CMC 담체는 점도가 큰 물질로 luminometer tube내의 사료에 유동을 최소화함으로써 산소의 공급을 일정하게 하여 bioluminecsence의 intensity를 일정하게 측정 할 수 있고, P phosphoreum을 고정하여 30분 후에 bioluminescence의 안정성을 가지므로 바로 측정할 수 있는 장점을 가지고 있다 독성에 민감하고 bioluminescence에 영향을 주지 않는 CMC의 농도(w/v)의 결정은 중요하며 1%(w/v) CMC 담체에 고정했을 때 bioluminescence의 유지가 가장 안정했으며, Cr 화합물에 대한 민감하게 반응하는 CMC 담체의 EC_50값ㅇ르 г값을 이용하여 산출했을 때 1% CMC 담체에 고정하였을 때 적은 독성 농도에서 민감한 bioluminescence의 감소를 볼 수 있었다. 그러므로 독성모니터용 P phosphoreun의 고정화 담체로 CMC의 1%(w/v)가 최적 농도이다. Bioluminescence of Photobacterium phosphoreum has been used for the detection of pollutants in the environment. Immobilization method was used to maintain the stability of bioluminescence of P. phosphoreum. Carboxymethylcellulose was investigated for immobilization of P. phosphoreum as a matrix without disturbance in bioluminescence emission. Maintenance of bioluminescence was determined from the P. phosphoreum immobilized on the various concentrations of carboxymethylcellulose. Relatively high bioluminescence intensity was observed with cells immobilized on 1%(w/v) carboxymethylcellulose. The effect of carboxymethylcellulose concentrations on the sensitivity of Cr-compounds including Na_(2)CrO_(4), K_(2)CrO_(4), CrO_(3), CrK(SO_(4))_(2) and CrCl_(3) to the bioluminescence intensity was investigated. The calculated EC_(50) showed that the linear relationship between such substances and bioluminescence.

Penicillium janthinellum의 사멸균체를 이용한 염료의 제거

이제혁,전억한 경희대학교 생명자원과학연구원 1997 硏究論文集 Vol.18 No.-

사멸균체를 사용한 염료의 흡착율은 배지환경의 pH 감소에 따라 증가하여 pH 2의 경우 염료 Appolocion H-E7B의 흡착ㄹ은 pH6에서 보다 약 60배의 초기 흡착율이 증가하였고, 온도는 높을수록 염료의 흡착율이 증가하여 실험 조건중 가장 높은 온도인 40℃에서 흡착ㄹ이 가장 양호하였다. 종래의 염색폐수 처리 방법으로 처리가 어려운 reactive염료의 흡착율이 우수하였고, 또한 혼합 염료도 흡착율이 양호하여 성상이 불균질한 염색 폐수의 처리에 적용 가능성이 긍정적이었다. 사멸균체에 detergent를 첨가할 경우 염료의 흡착이 저해되는데, 이것은 detergent가 균체의 염료의 결합 부위와 경쟁적으로 결합하여 흡착능을 저하시키는 것으로 사료되며, 각종 금속 ion의 존재시에도 사멸균체에 의한 염료의 흡착에는 거의 영향을 미치지 않아 각종 이온들이 복합적으로 함유된 염색폐수의 처리에도 Pen. janthinellum의 사멸균체가 적합한 것으로 사료된다. It is difficult to remove dyes which are major pollutants in the texile industries. The biosorption was carried out with dead biomass which were prepared by drying and grinding process of harvested microbial cells. The maximum biosorption was obtained with dead cells at pH 2.0 and 40 C. The initial biosorption rates were found to be 0.06, 0.086, 0.079(mg/gㆍmin) for each Apollocion Red H-E7B, Apollofix Red SF-3B, and Apollocion Red H-E3B, respectively. The effects of detergents and ions on the initial biosorption rates were also investigated to gain higher efficiency of dye removal since wastewater contains some ions and detergents. However, detergents added to the modified cells reduced dye biosorption rate and ions used also had no influence on the dye biosorption rates.

고정화 및 저장 온도에 따른 Photobacterium phosphoreum의 Bioluminescence 안정성의 변화

김현숙,정계훈,전억한 경희대학교 생명자원과학연구원 1998 硏究論文集 Vol.19 No.-

P phosphoreum의 고정화에 있어 중요한 것은 matrix의 선택이며, matrix로서 soudium alginate만을 사용하여 고정화 하는 것보다는 strontium chloride를 첨가하여 gel의 견고성을 높여주었을 때 세포의 bioluminescence 유지도가 증가하였다. 저장온도에 따른 세포의 bioluminescence 유지도와 활성도와 관련하여 -70℃, -20℃, 20℃에서 저장한 세포의 경우 저장 1일 후에 급격한 bioluminescence의 감소를 보였으며 낮은 세포 활성도를 보인 반면 4℃의 경우 bioluminescence의 유지도가 15일 이상 이어졌으며 높은 활성도를 나타내었다. 따라서 P phosphoreum의 bioluminescence 안정성에 있어 가장 좋은 결과를 나타낸 것은 2.5%(W/W) sodium alginated와 0.3M(W/V) strontium chloride로 고정화하여 4℃에 저장한 세포였다. The objective of this work was to unprove biolummescence stability of Photobacterium phosphoreum when stored at different temperature in view of developing contmuous on-line monitonng system for pollutants m environment. A long-term experiment was performed to determine how immobilization affects the mamtenance and stability of biolummescence from luminescent bacteria at appropnate temperature. The unmobhzed cells of P. phosphoreum were compared with free cells m terms of mamtenance of biolummescence at room temperature. It was found that the biolummescence of cells immobilized on strontium alginate showed higher biolummescence mtensity than both free and mixed cells with only algmate as a matrix. The effect of temperature on the biolummescence stability was investigated with free and immobilized cells stored at 20℃, 4℃, -20℃ and -70℃ for 20 days. Both free and immobilized cells stored at 4℃ emtted a stable biolummescence whde the biolummescence markedly decreased with those stored at 20℃, -20℃ and -70℃.

독성 물질 측정을 위한 Photobacterium phosphoreum의 고정화에 관한 연구

이은수,이홍주,전억한 경희대학교 생명자원과학연구원 1998 硏究論文集 Vol.19 No.-

독성 물질 측정용 P phosphoreum의 고정화 물질 선택을위하여 고정화 방법을 크게 4가지로 나누어서 그 방법에 따라 각각 고정화 물질 한가지씩을 선택하여 P phosphoreum의 bioluminescence 안정성을 알아보았다. Polyacrylamide나 collagen에서는 bioluminescence가 유지를 못하고 20분 안에 급격히 떨어졌으나, alginate와 k-carrageenan은 온도를 높여야 gel이 형성되는 성질을 갖고 있기 때문에 저온성 발광 미생물인 P phosphoreum에 적합한 matrix가 되지 못한다. 따라서 P phosphoreum의 biluminescence를 안정되게 유지하면서 고정화가 용이한 polymer로는 alginate가 적합한 고정화 물질로 선정되었고 독성물질에 대한 P phosphoreum의 민감성을 bioluminescence의 변화로 살펴본 결과 각각의 독성 물질마다 특이한 linearity한 관계식이 설정되었고 EC_(50)를 쉽게 구할 수 있었다. The bacterial bioluminescence has been studied to detect toxic materials m foods and environment. A monitonng system can be developed with a fact that the bioluminescence is mhibited sensitively when the bioluminescent bacteria are exposed to toxic substances. However, a stability of bioluminescence must be improved for the purpose of developing continuous monitoring system. The mmobilization of Photobacterium phosphoreum, therefore, was investigated m view of long term of bioluminescence stability. Various matrixes, sodium algmate, k-carrageenan, collagen and polyacrylamide were studied for proper immobilization matnx of P. phosphoreum, in order to improve stability of bioluminescence. Collagen and polyacrylamide were shown to be madequate matenals for immobilization of P. phosphoreum since the bioluminescence decreased when cells were mixed with such matnx due to toxic effect of free radicals of matnx. In case of k-carrageenan, the biolumexence was stable compared with collagen and polyacrylamide. However, the k-carrageenan was not also a suitable matnx because cells were not able to mix with the matnx properly m low temperature at which gel is formed and, especially, cells of P. phosphoreum are psychrophilic luminescent bacteria. With a sodium algmate, the bioluminescence was stably maintained for 20 minutes and, in addition to that, a process of immobilization was simple for bioluminescent bacteria. Therefore, by using a sodium algmate as an mobduahon matrix, the relationship between biolummescence of immobilized P. phosphoreum and each toxic substance was shown linearity and could calculate EC_(50).

염료제거능 곰팡이균주의 동정 및 염료 제거 특성에 관한 연구

이제혁,황규대,전억한 경희대학교 환경연구소 1995 環境硏究 論文集 Vol.6 No.-

F-2 isolates cultured on the agar plate containing the dyestuff has formed the clear zone around the strain. The morphology of F-2 was observed with microscope and identified as Penicillium janthinellum(subgenes Furcatum var. Divaricatum). The effects of pH on the absorbace of dyestuff almost not shown except above pH 12 and so the decrease of dye concentration during the incubation of Penjanthinellum is caused by the ability of Penjanthinellum that can remove the dyestuff. The pH and dry cell weight in the incubation of Penjanthinellum have changed after 18 hours and the decrease of the dye concentration in the medium has occured simultaneously. The rates of dye removal were good under the conditions of almost pH ranges except above pH 10 and 0.01g/100㎖ of dye was almost removed in over 3.0g/ℓ of starch concentration. In the case of l%(v/v) inoculum of Penjanthinellum in the 100㎖medium, 0.01g/100㎖ of dyestuff was almost removed in the 20 hours.

곰팡이균의 액내배양법에 의한 염료 분해

이제혁,황규대,전억한 慶熙大學校 1994 論文集 Vol.23 No.-

Dyes released into the environment from the textile industries are considered to be serious to cause polution problem. 12 strains of the dye-degradable fungi were isolated from the soil and strain showing high activity for biodegradation of dyes used for investigation was selected from 12 strains and coded as F-2. F-2 strain was grown in the medium containing dye as a submerged culture at pH 7∼8 and 30℃ and a dye investigated was degraded at high level(80%) within 48hrs. Comparatively mild agitation speed of impeller(150rpm) was used for the cultivation of fungi F-2.

PEG 분해균주의 분리와 PEG film의 상용성에 관한 연구

이제혁,정성제,이준열,전억한 경희대학교 부설 식량자원개발연구소 1993 硏究論文集 Vol.14 No.-

PEG를 sole carbon과 energy source로서 이용하는 미생물을 자연계에서 분리하였고, PEG의 분자량이 높아질수록 그 분해 미생물의 수가 감소하는 것을 확인하였다. 또한, liquid culture로서 PEG농도를 감소시키는 미생물을 선별하였고, 분해율은 PEG 8000이 약 18.8%였으며 PEG 10000은 약 25.4%인 것으로 조사되었다. PEG film의 제조를 위해 EMAA 및 EAA와의 상용성을 적외선 분광(IR) 스펙트럼을 사용하여 조사한 결과, EMAA와 EAA의 카르보닐기와 PEG의 에테르기와의 강한 수소결합이 형성으로 blend film제조시 상용성이 있는 것으로 확인되었다. Several strains capable of degrading PEGs(Polyethylene Glycols)were isolated and investigated for their biodgradation ability of PEGs. Microorganisms screened for the biodegradation studies were those grown on the PEG used as a sole carbon and energy source. It was known that the number of microorganisms decreased when grown on the high molecular weight of PEG(e.g. 20,000). A liquid culture was carried out with such microorgaisms and resulted in the decrease in PEG concentration meaning that PEG was degraded in the reactor. The biodegradability was found to be about 18.8% for PEG-8000 and 25.4% for PEG-10000, respectively. For the manufacture of biodegradable PEG film, EMAA/PEG and EAA/PEG blending ability was investigated with IR spectrum and showed that it was possible to produce blending film.

자연분해성 Polymer blend와 이 blend의 생분해에 관한 연구

이제혁,이준열,전억한 경희대학교 환경연구소 1993 環境硏究 論文集 Vol.4 No.-

Several strains capable, of degrading PEGs(Polyethylene Glycols) were isolated and studied for their biodegradation ability of PEGs. We have isolated the strains that used the PEG-compounds as a sole carbon and energy source. We have screened the strains that reduced the concentration of PEGs in the liquid culture. The biodegradability was about 18.8% in PEG-8000 and 25.4% in PEG-10000. In order to produce PEG-film, EMAA/PEG and EAA/PEG blending ability was tested with 1R spectrum. Therefore, the blend was possible to be produced. EMAA/PEG and EAA/PEG blend film were produced and tested on the biodegradation ability. As a result that was tested with ASTM method, it was found that some of the blend films have the biodegradability. And we have tested identification of cells used in this experiment. Cells were identified as Pseudomonas species and Flavobacterium species.