Gelatin 액화세균의 생장억제에 관한 연구

이기성,崔榮吉,趙賢淑 漢陽大學校 環境科學硏究所 1983 環境科學論文集 Vol.4 No.-

Gelatin 을 재료로 이용하는 식품의 폐기물에서 gelatin 을 액화시키는 능력이 특히 뛰어난 4종의 세균을 분리·동정하였으며, 배지내에 NaCI, KI·I₂, CuSO₄등의 억제제를 농도별로 처리했을때 나타나는 4균주의 생존도 및 액화능의 변화를 비교·검토하였고, 생존도에 미치는 pH의 영향도 조사하였다. 또한 MIC(Minimal Inhibitory Concentration) Test를 시행하여 항생제에 대한 4균주의 MIC도 조사하였다. 그 결과는 다음과 같다. 1. 4균주중 H strain은 Serratia liquefaciens로, Y₁strain은 Enterobacter속으로, Y₂와 Y₃strain 은 Pseudomonas 속으로 동정되었다. 2. 배지내에 억제제로 NaCl을 처리했을 경우, 7% 농도에서 생장이 100% 억제되었으며, KI·I₂처리시에는 20ppm 에서부터 생장이 억제되기 시작하여 100ppm에서는 4개의 균주 모두 100% 생장이 억제되었다. CuSO₄처리시에는 100ppm에서부터 억제효과를 나타내기 시작하여 300ppm에서는 거의 100% 억제 효과를 보였다. 생존도에 미치는 pH의 영향을 살펴본 결과, pH 5.0, 5.5, 6.0에서는 대조군과 동일한 생존도를 나타냈으나 pH 4.0, 4.5 에서는 100% 생장이 억제되었다. 분리 균주들의 액화능 비교에 있어서는 H strain 이 가장 뛰어난 액화능을 나타내었다. CuSO₄와 KI·I₂를 각각 3ppm, 10ppm의 저농도로 처리했을 경우 4균주의 액화능은 대체로 10∼20% 감소되었다. 3. 액화 능력이 가장 뛰어난 H strain 의 MIC 는 Penicillin G; 1∼5 ㎍/ml, Spectinomycin; 12.5∼25㎍/ml, Ampicillin; <0.05㎍/ml, Cefobid; <1㎍/ml, Claforan; 0.02∼0.3㎍/ml, Tetra-cycline; >10㎍/ml로 나타났다. Tetracycline을 제외한 5가지 항생제에 대하여 H strain이 Y strain들에 비해 민감한 것으로 나타났다. 그러나 Tetracycline의 경우에는 H strain이 Y strain들에 비하여 저항성을 갖는 것으로 나타났다. Four strains of gelatin-liquefying bacteria were isolated from the sewage of the food using gelatin as the material. Treated with various concentration of NaCl, KI-I₂and CuSO₄in the culture medium and with various gradient of pH, viability of the four strains was examined and compared with each other. And also, gelatin liquefying ability was investigated in the normal culture condition and in accordance with the concentration of the inhibitor. Morever, MIC(Minimal Inhibitory Concentration) test for these strains was also carried out. The results were as follows: 1. Of the four isolated strains, the H strain was identified as Serratia liquefaciens, Y₁as the genus Enterobacter, Y₂and Y₃strains as the genus Pseudomonas. 2. Treating the culture medium with NaCl as the growth inhibitor, 7% concentration of NaCl inhibited completely the viability of all the isolated strains. Adding KI-I₂solution to the medium at the level of 20ppm, viability began to decrease and the inhibitory effect was shown 100% at 100ppm. Treated with CuSO₄at the level of 10ppm, viability began to decrease, and at 300ppm growth inhibition showed almost 100%. There was no significant difference in viability according to the pH gradient at the range of 5.0-6.0. However, at pH 4.0 and 4.5, 100% inhibitory effect on the growth was shown. Considered with the ability of gelatin liquefaction, the H strain among the four strains had the most rapid and strong ability in gelatin liquefaction. (That is, about 23 times more effective in gelatin liquefaction than that of Y₁strain) 3. Considered the MIC test, the H strain was more sensitive to Penicillin G(1-5 ㎍/ml), Spectinomycin(12.5-25㎍/ml), Ampicillin(<0.05㎍/ml), Cefobid(<1㎍/ml) and Claforan (0.02-0.3㎍/ml) than to those of the strains. Compared with the other strains, the H strain was more insensitive up to the level of 10㎍/ml to the minimal inhibitory concentration of Tetracycline.

한양학원 폐수의 미생물학적 처리에 관한 연구

최용,최영길,이희준,안연준 漢陽大學校 環境科學硏究所 1994 環境科學論文集 Vol.15 No.-

한강수계인 안양천에서 유출되는 산업페수에서 카드뮴과 납에 내성을 지니며 이들 중금속을 축적하는 균주 HYM 2., HYM 40을 분리하여 동정한 효모인 Torulopsis inconspicua로 밝혀졌다. 납의 경우 180mg/g dried cell, 카드뮴의 경우 147.5mg/g dried cell까지 축적한는 것을 확인하였다. 전자현미경으로 관찰한 결과 이들 중금속은 주로 세포막에 축적되는 것으로 나타났다. 이 균주를 한양학원 폐수를 재료로 하여 pilot system에 적용한 결과 원폐수만을 적용했을 때보다는 활성슬러지를 첨가했을 때 (MLSS를 2000∼3000mg/ㅣ로 보정), 특히 분리된 균주를 적용시켰을 때 중금속의 제거 효율이 훨씬 더 증가하였다. 이는 환경오염원을 제어하는 균주를 직접 실공정에 적용할 수 있는 가의 가능성을 타진할 수 있었다. Two strains of yeasts, HYM 20 and HYM 40, which have resistance against heavy metals such as cadmium and lead and have ability to accumulate the heavy metal were isolated in the industrial sewage drained from Anyang stream. These strains of yeasts were identified as Torulopsis inconspicua. This yeast accumulated lead upto 180mg/g dried cell and cadmium 147.5mg/g dried cell. In the electron microscopic analysis, it was observed that the heavy metals were accumulated within the cell membrane. In the application of wastewater from Hanyang University to the A₂/0 system, increased accumulation was observed when the original sewage was added with active slvge by comparison with the sewage alone. particulaly, when added with sluge and isolated strains, higher increasemant of accumulation was boserved than in the former cases. These results showed the possibilities for the practical elimination of heavy metal pollutants.

酵母細胞의 인산대사 및 유기물 합성에 미치는 무기인산 화합물의 영향

崔榮吉,이기성,李鍾三,趙善姬 漢陽大學校 環境科學硏究所 1983 環境科學論文集 Vol.4 No.-

효모의 개체군 생장에 미치는 인산 화합물(??) 및 ??, ?? 양이온의 영향을 측정한 결과, ??는 Knopp씨 배지수준의 1배, 10배, 50배 처리의 전구간에 걸쳐서 개체군 생장에 유의한 촉진효과를 볼 수 없었다. ?? 처리구에서는, ?? 의 1배 처리구에서 대조구에 비하여 뚜렷한 생장촉진효과를 볼 수 있었으나, 10배, 50배의 처리구에서는 오히려 생장이 감소함을 나타내었다. 또한 ?? 처리구는 전 구간에 걸쳐서 대조구에 비하여 뚜렷한 억제가 관찰되었다. 한편, 효모의 인산화합물 합성에 미치는 영향을 측정하여 본 결과, ??의 경우는 산 가용성 Total-P 가 전 구간에 걸쳐 배양 처리후 6일째에, 대조구에 비하여 증가하였으며, ??처리구는 전처리구에서 산가용성 Total-P가 감소하였으나 ortho-P만은 증가하였음을 보였다. ??의 처리구에서도 Total-P는 역시 대조구에 비하여 감소하고 ortho-P는 대조구와 비슷한 경향을 보였다. poly-P "A", "B", "C", type 의 소장 관계를 측정한 결과, poly-p-A 와 B는 인산농도 및 양이온의 농도가 높을수록 많은 생성량을 나타내었으며, 특히 ?? ×50 처리구에서 poly-p-"B" 의 최대 합성량을 볼 수 있었다. 또한 여러 배양조건하에서도, 여러가지 poly-P 전환과정은 모든 시험구에서 동일한 것으로 나타났다. 그리고 특히 ?? 이온에 의해 영향받는 poly-P의 유기물합성에 이용되는 전환경로와 poly-P 상호 전환경로는 다음의 관계가 성립하는 것으로 추론된다. ?? 한편, 효모세포의 단백질, 핵산, 인지질, 탄수화물 합성에 미치는, 이들 인산 화합물의 양적 동태를 고찰한 결과, 핵산의 경우, ?? 처리구는 모든 구간에서 핵산 합성에 유의한 효과를 찾을 수 없었으며, ??처리구는 1배농도의 처리구에서는, 핵산의 합성증가를, 그외의 처리구에서는 합성량이 낮았다. ?? 처리구에서는 전체적으로 현저한 유기물의 합성억제를 나타내었다. 단백질 합성의 경우는 ??×1처리구를 제외한 모든 처리구에서 대조구와 비교하여 비슷한 경향을 나타내거나 억제효과를 보여주었을 뿐,촉진효과를 관찰할 수 없었다. 인지질 합성시 ??는 촉진효과를 나타내었으나 ?? 와 ?? 는 유의한 효과를 볼 수 없었다. 효모의 탄수화물 합성에 있어 ?? 는 대체로 대조구의 합성량과 유사하며, ?? 의 경우와 ??의 경우는 전 처리구간에서 대조구에 비하여 배양초기에 억제효과를 나타내었으나, 배양후기에는 오히려 증가되었다. 한편 poly-P와 유기물 합성과의 상관관계에서, Polysaccharide,lipid-p, DNA 합성에는 Poly-P-B가, nucleotidic-labile-P 생성에는 poly-P-C 가 상호 연관된 것으로 추정되었다. Examined the effect on the growth of yeast population by treating medium with phosphate compounds such as ??, ?? and ?? ion as the the concentration of Knopp's level, tenfold concentration and of fiftyfold, respectively, it could be recognized as the following facts; in case of ??, it showed no significant effect on the growth of yeast population in all treatmemt. And in case of ?? treatment replaced K? of KH₂PO₄in knopp's medium it showed the stimulating effect in the culture as comparing with those of control, but on the contrary, it was revealed the inhibition effect in the culture of tenfold and fifty fold treatment. However, the addition of ?? compounds showed the clear evidence of inhibition effect in population growth. Measuring the effect on the synthesis of phosphate compounds of yeast cell by treating the above chemicals, the amount of acid soluble-total phosphate was increased in all ?? teatment on 6th day after treatment. On the contraty, in case of ?? treatment, the amount of acid soluble-total phosphate was decreased in all treatment comparing with that of control, but the amount of ortho-phosphste was increased in all cultures. These similar tendencies were also found in the cultures of As?? compound treatment. The amount of acid soluble-total phosphate was decresed comparing with that of control and ortho-phosphate was as of control level. The accumulation amount of each type of polyphosphate(A,B and C) seemed to have a relationship between each other. It seemed that the amounts of polyphosphate A and B in yeast cell are increased in accordance with high cation concentration and phosphate concentration in culture medium. Especially, the culture in fiftyfold concentration of ?? ion synthesized the highest amount of polyphosphate B. However, the trun-over patterns of each polyphosphate under various conditions showed similar tendencies with each other. But the patterns utilizing in synthesis of cellular organic matter are belived to be affected by ?? ion as the direction in the following figure. ?? The synthesis of cellular organic matter, such as protein, nucleic acid, phospholipid , and polysaccharide was also belived to have a relation to cellular nutritional conditions. Any treatment of ?? showed no significant effect in synthesis of nucleic acid . The treatment of ?? compounds was resulted in decreased of nucleic acid generally, but increase in the culture replaced ?? of KH₂PO₄of Knopp's medium by ??. However, all the treatment of ??) compounds to medium showed strong inbibition effect in synthesis of nucleic acid. The synthesis of cellular protein is promoted by treating ?? compounds as Knopp's level. but in case of tenfold and fiftyfold concentration, the amount of protein is equivalent to or below to that of control. Referring to synthesis of phospholipid, ?? treatment was effective, but ?? and ?? compounds showed inbibition effect. The amount of carbohydrate in yeast cell treated by ?? is similar to those of control, however, in the cell treated by ?? or ?? compounds, the amount was increased just after decreasing in early stages of incubation. The relationship between each polyphosphate and synthesis of cellular organic matter could be interpretated that polyphosphate B is utilized mainly in synthesis of polysaccharide, phospholipid and DNA, and that polyphosphate C is also related in formation of nucleotidic labile phosphate.

極相林土壤과 土壞微生物의 分布

趙洪範,이기성,崔榮吉 漢陽大學校 環境科學硏究所 1985 環境科學論文集 Vol.6 No.-

경기도 광릉의 주엽산 일대의 삼림토양을 대상으로 5개의 정점을 설정하여 토양의 환경요인과 토양미생물의 군집규모를 조사하여 토양미생물의 토양생태계內의 기능을 토양의 환경요인과 상호 관련지어 해석하는 한편, 미생물의 개체군증식에 미치는 환경요인을 규명하는 미생물 생태학적인 연구를 실시하였다. 그리고 S/O value와 섬유소 분해 미생물 군집 크기 사이의 상관지수를 구하여, 토양생태계 內의 구조 및 기능 해석을 위한 새로운 biological index로서 S/O value가 적용 가능한 지를 조사하였다. 그 연구결과를 요약하면 다음과 같다. 1. 토양미생물의 군집 규모는 상층이 하층토양에 비하여 컸다. 또 토양미생물 군집의 규모를 세가지 유형으로 구분할 때, 일반세균>방선균류>일반균류의 순으로 군집의 크기가 확인되었으며 토양 內의 유기물 분해에 기여하는 작용도에 있어서도 그 순위가 동일하였다. 섬유소 분해능을 지닌 미생물 군집의 비율은 일반세균의 경우 75.8%, 방선균류는 81.2%, 일반균류는 54.5% 정도가 되어 매우 높았다. 한편 세균류와 균류의 개체군 규모는 3:1의 비를 나타내었다. 2. 실험대상지역의 토양 pH는 5.2∼5.7범위였으며, 토양 온도 및 무기이온의 함량은 일반적으로 상층이 높은 값을 나타내었으나 함수량은 하층이 높은 값을 보였다. 환경요인중 개체군 증식에 기여하는 영향이 가장 큰 parameter는 pH로 추정되었는데, 함수량, ??이온의 함량도 어느정도 영향력을 미치는 것으로 추정되었다. 3. 환경요인의 분석에서 수용성 당류, 아미노산의 함량은 상층이 하층에 비해 높았으나 총 유기물함량은 수직적으로나 정점별로도 유의한 차이는 없었다. 4. S/O value는 개체군 규모가 클수록 높았다. S/O value와 섬유소 분해 미생물 군집의 크기 사이의 상관지수는, S/O value와 총 섬유소 분해 미생물 사이에서 0.769, 일반 섬유소 분해 세균과의 경우 0.700, 방선균류 0.788, 일반균류는 0.576으로 나타나, 상당히 유의한 상관관계가 있었다. 5. 토양의 textile을 예견할 수 있는 균사체 발달 정도는 천이과정 중의 정점에서, 극상단계에 거의 도달한 정점에 비하여 양호하였다. 6. 실험대상지역의 토양생태계는 생물군집 중 미생물 군집의 규모 수준에서 고찰해 볼 때 극상단계에 도달한 것으로 판단된 정점도 실질적으로 아직 천이과정 중인 생태계로 판명되었다. The authors surveyed around Mt. Choo Yub in Kwang Reung on April 14th in 1983 and examined both environmental factors and microbial population size including the cellulolytic microbes in soil. In order to elucidate the relationship between soil environmental factors and the function of soil microbes in the soil, and to examine the environmental factors affecting to the microbial population size, this research was carried out. And calculated the correlation coefficients between S/O value and population size of cellulolytic microbes, S/O value was examined to be applied as new biological index for analysis of structure and function in soil ecosystem. The results of this experiment are summarized as follows. (1) The population size of soil microorgasms in surface layer was larger than that of lower layer. Subdivided the microbial community in the soil into three groups such as general bacteria, actinomycetes and general fungi, the population sizes and the activity in degradation of organic substances were large in order of general bacteria, actinomycetes and general fungi, respectively. Composition of cellulolytic microbes in subdivided microbial group was high as much as 75% in bacteria, 81.2% in actinomycetes and as 54.5% in fungi. The ratio of bacteria to fungi was 3:1. (2) PH value of the experimental soil was generally the range of 5.2∼5.7, the temperature of surface layer was higher about 5℃than that of lower layer. The amounts of inorganic components such as ??, ??, ??, ??, ?? were more accumulated in surface soil than those of lower layer. It is postulated that the parameter affecting strongly to the population size is pH among the various environmental factors. In addition, the moisture content and the amount of ?? were also related with the population size of soil microbes, to some extent. (3) In the analysis of organic substances such as free amino acid, soluble sugar and total organic matter, the amount of free amino acid and soluble sugar in surface layer were higher than those of lower later, but the amount of total organic matter has no significance in the sense of amounts between surface layer and lower layer. (4) The S/O value(amount of soluble sugar/amount of total organic matter) in soil was high in accordance with increase of the population size. And the correlation coefficients between S/O ratio and population size of cellulolytic microbes were shown such as 0.769, 0.700, 0.788, 0.576 respectively, in total cellulolytic population size, general bacteria, actinomycetes and general fungi. That is to say, it is significantly correlated between S/O ratio and population size of cellulolytic microbes. It was considered that S/O value can be adapted as a useful biological index for interpretation of the function and structure in soil ecosystem. (5) Mycelial extension to predict the textile of soil is more active in successional ecosystem than in the almost matured climax ecosystem. (6) The ecosystem of experimental area referring to the level of microbial population size experimental area is actually postulated successional ecosystem.

카드뮴내성군주의 카드뮴 적응기에 나타나는 생리생화학적 변화

김영호,김광훈,이기성,최영길 漢陽大學校 環境科學硏究所 1991 環境科學論文集 Vol.12 No.-

산업폐수에서 분리한 Bacillus sp.를 카드뮴 첨가배지에서 배양하면 2mM 농도까지 내성을 가지며, 세포내 무기인산 중합체의 축적이 증가되는 바, 이들 무기인산의 대사에 관여하는 제반 세포내 생화학적인 변화양상을 조사하고자 하였다. 카드뮴 첨가 배양에 따른 효소 활성도는 poly-Pase 와 ACPase는 상당히 증가되었지만 ALPase 는 그 활성이 카드뮴의 영향으로 감소되었다. 카드뮴 첨가 배양에 따른 세포의 인지질의 변화 양상을 살펴 보면 phosphatidyl glycerol 에 대한 phosphatidyl choline의 상대적 비율이 lyso-phosphatidyl choline의 상대적 비율보다 훨씬 많게 관찰되어 phosphatidyl choline 이 lysophosphatidyl choline으로 분해되는 경로가 카드뮴에 의해 억제되는 것으로 사료된다. 한편, nucleotide pool의 변화 양상을 살펴 보면 카드뮴의 영향으로 adenylate pool을 증가하며 카드뮴 비처리 복합 배지에서 배양한 세포에서는 cytidylate pool이 현저한 증가를 보인 반면 starvation(카드뮴을 첨가하지 않은 당과 인산 결핍 배양)시킨 세포에서는 guanylate pool이 증가됨을 관찰할 수 있었다. Upon cadmium added medium, in the course of culturing Bacillus sp. which was isolated from sewage of Daejon industrial plants and which is cadmium resistant up to the concentration of 2mM, we observed that inorganic polyphosphate was highly accumulated. In order to elucidate regulatory mode of this phenomenon, we tried to check biochemical changes which were correlated with phosphate metabolism ; such as phosphatase activity, phospholipid pattern and nucleotide pool. It found that, depending upon culture with added cadium, enzyme activity was markedly increased in poly-Pase, and in ACPase also, but the effect of cadmium resulted in a decrease in ALPase activity. The variation of cellular content of phospholipids with cadmium addition was showed that relative ratio of phosphatidyl choline for phosphatidyl glycerol was major to that of lysophosphatidyl choline for phosphatidyl glycerol, implicating degradation pathway of phosphatidyl choline into lysophosphatidyl choline was repressed by cadmium. On the order hand, with respect to the change of nucleotide pool, it should be noted that the effect of cadmium induced a increase in cytidylate pool, which considerly increased in Cd-free complex medium, when compared with the starvation of a culture of cells with sugar-phosphate free medium without added cadmium.

Photosynthetic artificial organelles sustain and control ATP-dependent reactions in a protocellular system

Lee, Keel Yong,Park, Sung-Jin,Lee, Keon Ah,Kim, Se-Hwan,Kim, Heeyeon,Meroz, Yasmine,Mahadevan, L,Jung, Kwang-Hwan,Ahn, Tae Kyu,Parker, Kevin Kit,Shin, Kwanwoo Nature Publishing Group, a division of Macmillan P 2018 Nature biotechnology Vol.36 No.6

<P>Inside cells, complex metabolic reactions are distributed across the modular compartments of organelles(1,2). Reactions in organelles have been recapitulated in vitro by reconstituting functional protein machineries into membrane systems(3-5). However, maintaining and controlling these reactions is challenging. Here we designed, built, and tested a switchable, light-harvesting organelle that provides both a sustainable energy source and a means of directing intravesicular reactions. An ATP (ATP) synthase and two photoconverters (plant-derived photosystem II and bacteriaderived proteorhodopsin) enable ATP synthesis. Independent optical activation of the two photoconverters allows dynamic control of ATP synthesis: red light facilitates and green light impedes ATP synthesis. We encapsulated the photosynthetic organelles in a giant vesicle to form a protocellular system and demonstrated optical control of two ATP-dependent reactions, carbon fixation and actin polymerization, with the latter altering outer vesicle morphology. Switchable photosynthetic organelles may enable the development of biomimetic vesicle systems with regulatory networks that exhibit homeostasis and complex cellular behaviors.</P>

Biohybrid Swimmers Developed from Human Stem Cell-Derived Cardiomyocytes

Keel Yong LEE,Sung-Jin PARK,David G. MATTHEWS,Sean L. KIM,Carlos Antonio MARQUEZ,John F. ZIMMERMAN,Herdeline Ann M. ARDOnA,Andre G. KLEBER,George V. LAUDER,Kevin Kit PARKER 한국생물공학회 2022 한국생물공학회 학술대회 Vol.2022 No.9

Anti-hyperglycemic and Anti-hyperlipidemic Activities of Acanthopanax Senticosus Herbal Acupunture in C57BL/6J ob/ob Mice

Lee Sang-keel,Kim Yong-suk,Kang Sung-keel 대한침구의학회 2006 대한침구의학회지 Vol.23 No.2

수계 종속영양세균의 소장에 미치는 Azo 계 염료의 영향 및 염료 분해미생물의 분리

이용진,조홍범,최영길,이호용 ( Yong Jin Lee,Hong Bum Cho,Yong Keel Choi,Ho Yong Lee ) 한국하천호수학회 1993 생태와 환경 Vol.26 No.2

Effects of Orange II, a kind of azo dye, on heterotrophic bacterial population in Han river were not significantly toxic. Survival rate of the heterotrophic bacteria in each site was shown 22.85-51.31% at 5000 mg/l of Orange II. LD_50 of Orange II are 1000 mg/l or more in each site. Heterotrophic bacteria capable of degrading Orange II as a sole carbon source were not isolated in Han river but isoated Torulopsis sp., a kind of yeast, in wastewater of textile industry. The degradation rate of this isolate was over 90% after 66 hours culture in PAS medium containing 10 mg/l of Orange II and 2% of glucose. Its optimum temperature and pH are 37˚C, pH 7, respectively.

Saccharomyces uvarum 의 Catabolic Repression 시기에 유도되는 Ribosomal Ribonuclease 에 대한 연구

이기성,최영길,윤성녀 한국균학회 1986 韓國菌學會誌 Vol.14 No.3

In order to study subcellular locality and characteristics of ribonuclease in Saccharomyces uvarum, subcelllar fractions 45,000 x g pellet fraction, post ribosomal fraction and ribosome fraction were extracted during late log, stationary phase and sugar starvation conditions. Ribonuclease activity was significantly increased in ribosomal fraction under stationary and sugar starvation conditions. Ribosomal ribonuclease was extracted by EDTA plus streptomycin sulfate sad ammonium sulfate precipitation. The amount of ribosome in stationary and sugar starvation condition was decreased three to six fold as compared to that in the early log phase. The end products of ribosomal ribonuclease were detected by thin layer chromatography. It is postulated that the increase of ribosomal ribonuclease activity under sugar starvation results from 5'-rRNase, while the increase of rRNase activity under stationary phase results from 3'-rRNase.