남조분해 균류의 분리와 동정

최영길,민병례 한양대학교 자연과학연구소 2001 自然科學論文集 Vol.20 No.-

부영양화 수역의 4개 저수지로부터 곰팡이 12균주를 분리하였으며, 이들 분리 곰팡이들 중 남조류의 생장억제 능력이 우수한 HCLF-34와 HCLF-21을 왕송저수지로부터 분리하였다. 형태적인 특징에 따라 Penicillium sp.(HCLF-34)와 Acremonium sp.(HCLF-21)로 동정하였다. 두 개의 분리 곰팡이중 Penicillum sp.(HCLF-34)만이 Anabaena cylindrical lawn과 M.luteus agar p;ate에서 균체, 배양상등액 및 멸균된 배양상등액에서 plaque를 형성하였다. A.cylindrica와 Penicillium sp.(HCLF-34)의 혼합배양시 배양 3일 후 남조류의 분절 및 sheath의 손상이 관찰되었고, A.cylindrica와 Penicillium sp.(HCLF-34) 혼합배양시 배양 10분 후부터 남조류의 분절 및 sheath의 손상이 관찰되었다. The fungi that hydrolyze the blue green algae were isolated from Wangsong reservoir shown in eutrophication and identified as the genera Acremoniwm sp., and Penicillium sp. By morphological characters. The Penicillium. sp.generally showed less lytic activity than the Acremonium sp. on the Anabaena cylindrical agar lawn, but the cultured supermatant of Penicillium sp. Exhibited greater activity than those of the Acremonium sp. On Micrococcus luteus. In the mixed culture of bule green algae(Anabaena cylindrical) and the fungi(HCLF-34), the blue green algae was segmented and lysed in 3 days. In addition, the blue green algae(Anabaena cylindrical) was segmented and the width sheath of Anabaena cylindrical was appeared as lysed on the scanning electron microscopy, in mixed culture between the algae and fungi.

Cyanophage에 의한 남조류의 성장 억제효과

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

Cyanophage를 처리하였을 때 숙주세포의 성장율이 전반적으로 억제되었다. BG-11 배지를 이용하여 cyanophage : cyanobacteria의 비율을 1 : 1로 넣어주었을 때와 대수 증식기 중기의 숙주 배양액에 cyanophage를 처리하였을 때 가장 높은 남조류의 성장 억제효과를 가져왔다. 또한 대수 증식기 말기와 정지기에 cyanophage를 처리한 경우 숙주세포의 성장이 oscillation 현상을 보여 숙주와 virus간의 길항적 관계에 의한 성장조절의 가능성을 시사하였다. Cyanophage suppressed the growth rate of host cyanobacteria. The suppression effect was maximal when cyanophage was added with the ratio of cyanophage : cyanobacteria, 1 : 1 and on the midexponetial phase of host. Treatment of cyanophage on the stationary phase of host resulted in oscillation of host population and its cyanophage, which suggests the possibility of biological control by the antagonistic relationship of host-cyanophage system.

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.

유치원 안전관리에 대한 교사의 지각 연구

박용길 국립7개대학공동논문집간행위원회 2004 人文學論叢 Vol.4 No.-

The purpose of this study was to examine the real condition of safety management in kindergarten. This research was classified in the actual condition of safety management(safety inspection, emergency communication, institution equipments, indoor safely, outdoor safety, field learning) in kindergarten. Questionnaire is adopted to obtain data required for pertinent analysis. The subjects were 213 kindergarten teachers located in Choongnam and Daejeon province. SPSS 11.5 for Windows Program was utilized to calculate percentage, mean and standard deviation. Also, t-test, One-way ANOVA and Duncan's multiple range, were used. The main results of this study are as follows. There was significant difference in the actual condition of safety management in kindergarten according to demographic variables. Specially, there was difference of safety education according to teachers' position, age, and teaching experience. The importance of safety education must be emphasized. Coming up with the ways to teach and developing safety management program are needed urgently.

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.

자외선 살균효과에 미치는 옥수(玉水)와 옥분말(玉粉末)의 영향

최영길,민병례 한양대학교 자연과학연구소 2001 自然科學論文集 Vol.20 No.-

생체에 대한 옥수와 옥분말의 자외선 차단효과를 알아보기 위하여 동물의 피부 생태계와 연관있는 미생물들(Gram+, Gram-, 및 Eucaryotic yeast)을 대상으로 실험한 결과 다음과 같은 결과를 얻었다. 1.자외선 조사후 생존한 미생물 개체수의 측정결과로 분석해 볼 때 옥수첨가 액체배지에서 보다 옥분말 첨가 액체배지에서 자외선 차단효과가 높게 나타났다. 2.자외선 차단효과는 Escherichia coli, Saccharomyces cerevisiae, Bacillus subtilis의 배양체의 순으로 확인되었다. This experiment was designed to elucidate the effect of jade product (jade powder and jade water) on the sterilization by ultraviolet ray as the ecological model of animal skin. The microorganisms(Gram+, Gram-, and Eucaryote) which are closely related to animal skin were cultured in broth medium with jade powder and jade water, illuminated by ultraviolet ray to count viable number of microorganism, and evaluated the effect of jade product on the strength of ultraviolet ray. The results of this experiment are summarized as the followings. 1.The survival number of microbes illuminated by UV light were higher in the cultures of jade powder than those in the jade water, It could be evaluated that jade powder has an effect to interrupt the sterilization of UV light. 2.The sterilization by ultraviolet ray was decreased in order of the cultures as the Escherichia coli, Saccharomyces cerevisiae, and Bacillus subtilis.

Pseudomonas aeruginosa 의 고온 충격 반응

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

최적의 성장 온도가 37℃인 Pseudomonas aeruginosa를 고온 충격(heat shock)을 주기 위하여 45℃와 48℃에서 각각 배양하였고, 이때 생성된 단백질들의 조성을 sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)를 이용하여 37℃에서 합성된 단백질들의 조성과 비교하였다. 실험을 수행한 45℃와 48℃에서 자란 균주가 37℃에서 자란 균주보다 적어도 3종류의 단백질(60kD, 68kD, 그리고 70kD)들을 더 많이 생성하였고 이들 단백질들은 고온에 의하여 생성된 잘못된 기능을 가진 단백질들을 세포내에서 제거하는 역할을 하고 있다. 같은 조건에 전사단계를 억제한 rifampicin 을 처리한 결과, 이들 단백질의 생성은 증가하지 않았다. 이상의 결과로 부터 45℃와 48℃에서 합성이 증가된 60kD, 68kD, 그리고 70kD의 단백질은 P. aeruginosa에서 고온 충격에 의해 전사단계에서 합성이 조절되는 고온 충격 단백질들임을 확인하였다. P. aeruginosa grows optimally at 37℃. When this strain has transferred to 45℃ or 48℃, the growth of the cell has been affected by the high temperature. Especially, some proteins(60kD, 68kD, and 70kD) were observed that those proteins were overexpressed to eliminate the malfunctional proteins, which were produced by high temperature in the cell. When the cell were grown at 45℃ or 48℃ with rifampicin, which is the transcription inhibitor, those effects were not observed. From these results, proteins produced from the culture conditions of 45℃ or 48℃ are clarified to be heat shock proteins which are regulated by the transcriptional level.

유독 중금속 오염물질 처리를 위한 미생물균주의 최근 이용 및 개발

방상원,최영길,한명수 한국환경생물학회 2001 환경생물 : 환경생물학회지 Vol.19 No.2

There are several ways to remove and treat toxic heavy metals in the environment: chemical, physical and biological ways. The biological treatment utilizes the natural reactions of microorganisms living in the environments. These reactions include biosorption and bioaccumulation, oxidation and reduction, methylation and demethylation, metal-organic complexation and insoluble complex formation. The biological reactions provide a crucial key technology in the remediation of heavy metal-contaminated soils and waters. According to recent reports, various kinds of heavy metal species were removed by microorganisms and the new approaches and removal conditions to remediate the metals were also tried and reported elsewhere. This was mostly carried out by microorganisms such as fungi, bacteria and alga. In addition, a recent development of molecular biology shed light on the enhancing the microorganism's natural remediation capability as well as improving the current biological treatment.

Gamma선 조사에 의한 식품미생물의 살균효과

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

한국산 건강식품 가공원료인 알로에, 화분, 효모, 효소 가공분말의 초기 미생물오염도는 각각 8.2× 10²cell/g, 8.7 ×??cell/g, 8.8 ×10⁴cell/g, 6.9× 10⁴cell/g으로 측정되었다. 각 시료군별 감마선 조사를 1∼9kGy로 적용한 결과 초기 미생물의 오염도가 가장 낮은 알로에 분말의 경우 3.5kGy에서 완전 살균이 나타났으며 초기 오염도가 가장 높은 화분 가공분말은 7.2kGy정도에서 완전 살균이 가능하였다. 알로에, 화분, 효모 및 효소 가공분말의 선량 적용에 따른 미생물 생존 곡선에서 ??값은 각각 1.30, 1.20,1.27, 1.20으로 산출되었으며 불활성계수(n)가 6일때 7∼8kGy의 치사선량(??)값을 얻었다. The sample used in this experiment was worked material of health foods manufactured in Korea such as aloe, pollens, yeast prepns. and enzyme prepns. The initial microbial population of above food was 8.2 ×10²cell/g-aloe powder, 8.7 ×??cell/g -pollen, 8.8× 10⁴cell/g-yeast prepns, 6.9 ×10⁴cell/g-enzyme prepns. When samples were irradiated as a serial dose from 1kGy to 9kGy using gamma ray-??, aloe powder showed complete sterilization under 3.5kGy dose. In contrast, complete radio-sterilization in pollens is possible around te 7.2kGy dose of ??-gamma irradiation. ?? value(the dose required to eradicate 90% of the population)presented in microbial suvival curve according to serial irradiation dose was calculated 1.30(aloe), 1.20(pollens), 1.27(yeasts prepns.) and 1.20 (Enzymes prepns), respectively. As a inactivation factor(n) was 6, Lethal Dose(??)is 7∼8kGy dose of gamma irradiation.

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.