Aureobasidium pullulans IMS-822가 생산하는 ${\beta}-(1{\to}3)(1{\to}6)-Glucan$의 특성 분석

이석준,안극현,박찬선,윤병대,김민수,Lee, Seog-June,Ahn, Keug-Hyun,Park, Chan-Sun,Yoon, Byung-Dae,Kim, Min-Soo 한국미생물학회 2009 미생물학회지 Vol.45 No.1

국내 토양시료로부터 점액성 고분자물질을 생산하는 효모 균주를 분리 및 동정하여 Aureobasidium pullulans IMS-822라 명명하였다. A. pullulans IMS-822가 생산하는 외분비성 폴리머를 정제 및 동결 건조하여 구조분석을 수행하였다. HPLC를 이용한 구성당 성분분석에서 포도당 단일성분으로 구성되어 있음을 확인하였다. FT-IR을 이용한 구조분석에서는 890 $cm^{-1}$ 부근에서의 ${\beta}-configuration$을 확인하였으며, 3300 $cm^{-1}$ 부근으로 shift 된 완만한 -OH stretching은 폴리머를 구성하고 있는 분자 사이에 강한 수소결합이 작용하고 있는 것으로 추측되었다. $^{13}C-NMR$ 분석결과는 A. pullulans IMS-822가 생산하는 외분비형 다당류인 (1,3)-linked ${\beta}-D-glucosyl$ unit의 6개의 탄소, ${\delta}$ 105.05 ppm (C1), 75.82 ppm (C2), 86.62 ppm (C3), 70.61 ppm (C4), 78.13 ppm (C5), 63.22 ppm (C6) 및 ${\beta}-(1,3)D-glucosyl$ 잔기에 분기사슬 되어 있는 ${\delta}$ 72.11 ppm의 C-6의 signal을 확인할 수 있었다. A. pullulans IMS-822가 생산하는 ${\beta}-(1{\to}3)(1{\to}6)-glucan$의 알칼리 수용액(0.5%, w/v)에 24.0 ${\mu}M$의 Congo red 첨가하여 30분간 반응시킨 후 $400{\sim}700$ nm의 범위에서 최대흡수파장의 변화를 조사한 결과 NaOH의 농도가 증가함에 따라 최대흡수파장은 장파장으로 이동하여 498 nm에서 515 nm까지 이동하였다. 최대흡수파장은 0.4 M NaOH 농도에서 점차로 감소하여 506 nm 부근에서 안정화되는 것으로 나타남으로써 순차적 구조를 가지는 것으로 확인되었다. The exo-polysaccharide producing microorganism, Aureobasidium pullulans IMS-822, was isolated and identified from soil. The viscosity-average molecular weight of exo-polysaccharide was calculated as $8.9{\times}10^5$ by Mark-Houwink equation. The sugar component of exo-polysaccharide was determined as glucose by HPLC analysis. The IR spectra indicated that the exo-polysaccharide has an absorption peak at 890 $cm^{-1}$ for the ${\beta}-configuration$ of D-glucan. The $^{13}C$ NMR signal at ${\delta}$ 86.62 ppm arose from the substituted C-3 of glucose. The signal at ${\delta}$ 72.11 ppm was assigned to C-6 of branched ${\beta}-(1{\to}3)-D-glucosyl$ residues. Viscosity and Congo red reaction indicated that {\beta}-(1{\to}3)(1{\to}6)-glucan$ produced by A. pullulans IMS-822 has a highly ordered hydrogen-bond dependent conformation in aqueous solution, which collapses in strong alkaline solution.

식품 및 환경 , 기타 축산폐수의 고도처리 및 지질생산을 위한 Botryococcus braunii 의 대량배양

이석준(Seog June Lee),김성빈(Seong Bin Kim),김희식(Hee Sik Kim),권기석(Gi Seok Kwon),윤병대(Byung Dae Yoon),오희목(Hee Mock Oh) 한국미생물생명공학회 1999 한국미생물·생명공학회지 Vol.27 No.2

N/A

제초제 paraquat의 토양중 흡ㆍ탈착 특성

이석준(Seog June Lee),김병하(Byung-Ha Kim),김장억(Jang-Eok Kim) 한국농약과학회 1998 농약과학회지 Vol.2 No.1

규산질다공체와 미생물응집제의 녹조제어 효과

박명환(Myung Hwan Park),이석준(Seog June Lee),윤병대(Byung Dae Yoon),오희목(Hee Mock Oh) 한국환경생물학회 2001 환경생물 : 환경생물학회지 Vol.19 No.2

N/A The effects of CellCaSi and bioflocculant on the control of algal bloom were investigated in enclosures in a small eutrophic pond. The bioflocculant produced by a bacterial strain S-2 was finally selected to remove Microcystis aeruginosa which was a dominant species of algal bloom in the pond. Enclosure experiment showed that phosphorus concentration decreased dramatically from 131㎍l^-1(Control) to 1∼14㎍l^-1 in three CellCaSi-enriched enclosures. Chlorophyll-α concentration also decreased from 215㎍l^-1(Control) to 59㎍l^-1 by the addition of CellCaSi(1gl^1), bioflocculant(2ml l^-1), calcium chloride(1gl^1) and ferric chloride(2㎎ Fe l^-1) in Enclosure 4. From the results of the mouse acute toxicity test of the S-2 biofloccculant and the goldfish survival test in enclosures, it seems that both the S-2 bioflocculant and the CellCaSi do not show any severe toxicity in water system. Consequently, it was concluded that the bioflocculant and the CellCaSi could be used to control algal bloom in eutrophic waters by removing phosphorus and chlorophyll-α.

대청호 성층파괴시 cyanobacteria 의 수직분포 및 세포내 인의 조성

이희목(Hee Mock Oh),이석준(Seog June Lee),박정호(Jung Ho Park),맹주선(JueSon Maeng) 한국하천호수학회 1997 생태와 환경 Vol.31 No.1

N/A

Algal bioassay 에 의한 조류생장 제한영양염류 결정

오희목(Hee Mock Oh),이석준(Seog June Lee),김성빈(Seong Bin Kim),박미경(Mi Kyung Park),윤병대(Byung Dae Yoon),김도한(Do Han Kim) 한국하천호수학회 1998 생태와 환경 Vol.31 No.2

Determination of Limiting Nutrient for Algal Growth by Algal Bioassay. Oh, Hee-Mock, Seog June Lee, Seong-Bin Kim, Mi- Kyung Park, Byung-Dae Yoon and Do-Han Kim (Environmental Microbiology Research Unit, Korea Research Institute of Bioscience and Biotechnology, Taejon 305-333, Korea and Water Resources Research Institute, Korea Water Resources Corporation, Taejon 305-390, Korea) Water quaiity and algal growth potential (AGP) using Microcystis aeruginosa were investigated from April to September 1997 in Daechung Reservoir. Secchi disk transparency was 1.1 m in August. Chlorophyll-a concentrations were higher than 10㎍/l at all samples. Total phosphorus concentration was in a range of 0.020∼0.027mg/l which is at the approximate value of 0.03 mg/l, the general critical level of eutrophication. These results indicate that Daechung Reservoir is in an eutrophic state except some temporary times and restricted sites. Phosphorus uptake rate of M. aeruginosa was higher in the water from the epilimnion than in the hypolimnion and also higher under light condition than under dark condition. Therefore, phosphorus uptake rate appears to depend on cellular phosphorous content and light condition, a meaning of endergonic process. Phosphorus was the most frequently limiting nutrient: phosphorus was limiting algal gowth in April and May, nitrogen, phosphorus and trace elements were the limiting factors for algae in August, and trace elements in September. If a fast growing alga, M, aeruginosa, is used as a test organism for assaying AGP, in vivo fluorescence instead of dry weight can be measured for algal biomass and incubation period can be reduced from traditional 14 days to 7 days.

규산질다공체와 무기첨가물의 수중 인 제거 효과

박명환(Myung Hwan Park),한명수(Myung Soo Han),이석준(Seog June Lee),안치용(Chi Yong Ahn),윤병대(Byung Dae Yoon),오희목(Hee Mock Oh) 한국하천호수학회 2002 생태와 환경 Vol.35 No.3

The CellCaSi, a porous silicate material, was tested for the removal of phosphorus(Pas phosphate) n water. The effect of the CellCaSi was investigated on the basis of both particle size(under 1, 2 and 4㎜) and added amount(0, 1, 2.5, 5, and 10g/ℓ) of the CellCaSi. The removal efficiency of phosphorus was highest with a particle size of under 1㎜ and also increased with an increasing amount of the CellCaSi. The pH change showed little effect on the phosphorus removal of the CellCaSi. The calcium ion was eluted from the CellCaSi into the water, while the aluminium and iron were not. The eluted calcium ion was combined with dissolved phosphorus and then precipitated. The highest removal efficiency of phosphorus was obtained by the combined addition of the CellCaSi, calcium chloride, and ferric chloride. That is, the phosphorus concentrations of 0.10 and 1.0 ㎎/ℓdecreased to 0.03 and 0.47 ㎎/ℓ by the addition of the CellCaSi(1 g/ℓ), calcium ion(30 ㎎/ℓ), and ferric ion(1 ㎎/ℓ) at day 8 after treatment. The water qualities at the end of the experiment were as follows:pH was 8.1 and conductivity was 318 ??S/㎝ for raw and potable waters).

축산폐수의 3차처리를 위한 미세조류의 선별 및 처리조건의 최적화

김성빈,이석준,김치경,권기석,윤병대,오희목 한국산업미생물학회 1998 한국미생물·생명공학회지 Vol.26 No.1

축산폐수는 과량의 질소와 인을 함유하고 있어서 호소의 부영양화로 수질을 악화시키기 때문에, 본 연구에서는 2차처리된 돈폐수를 대상으로 조류를 이용하여 폐수 내 질소와 인을 효율적으로 제거하고자 조류의 선별 및 배양의 최적화 실험을 수행하였다. Chlorella vulgaris(UTEX 265). Chlorella sp. GE 21, Botryococcus braunii(UTEX 572), Botryococcus sp. GE 24, Scenedesmus quadricauda, Phormidium sp. GE 2, 그리고 Spirulina maxima(UTEX 2342)의 7종을 대상으로 조사한 결과 C. vulgaris(UTEX 265)가 질소와 인의 제거 뿐만 아니라 미세조류의 생산량도 우수하였다. 다음 실험에 사용될 미세조류로 선별된 C. vulgaris는 수온 35℃에서 생장이 가장 좋았으며, 약 알칼리인 초기 pH 8.0에서 생장이 우수하였다. 미세조류의 생산 및 질소와 인의 제거에 있어 광도의 영향을 조사한 결과 40μE/㎡/s 이하에서는 조류의 생장이 억제 받았으나, 그 이상의 광도에서는 비슷한 결과를 얻었다. 시료로 사용된 돈폐수의 질소와 인 농도는 각기 58.7 ㎎/ℓ와 14.7 ㎎/ℓ로 조사되었으며, 대상 폐수를 75%(돈폐수:지하수=3:1)로 희석하였을 때 질소와 인의 제거효율과 미세조류의 생산이 우수하였고, 희석하지 않은 경우 lag time은 길어졌으나 점차로 질소와 인이 제거되었다. 본 실험에서 얻어진 최적 조건하에서 25%의 돈폐수를 처리한 결과 4일 후 폐수중의 총질소는 95.3%가 총인은 96.0%가 제거되었다. The feasibility of algae as means of removing nitrogen and phosphorus from secondary treated swine wastewater was studied. Among the tested 7 species of Chlorella vulgaris (UTEX 265), Chlorella sp. GE 21, Botryococcus braunii (UTEX 572), Botryococcus sp. GE 24, Scenedesmus quadricauda, Phormidium sp. GE 2, and Spirulina maxima (UTEX 2342), C. vulgaris was selected for its fast growth and abilities to remove nitrogen and phosphorus and to produce algal biomass from swine wastewater. C. vulgaris grew well at 35℃, and the optimum initial pH for growth was 8.0. In the effect of light intensity, the growth of C. vulgaris was limited under a light intensity of less than 40μE/㎡/s. The secondary treated swine wastewater contained 58.7 ㎎/ℓof total nitrogen and 14.7 ㎎/ℓof total phosphorus, and was diluted to 75, 50, and 25% with groundwater to be treated. Nitrogen and phosphorus were removed by C. vulgaris in all diluted swine wastewaters among which the most effective removal was in 75% swine wastewater (swine wastewater:groundwater=3:1). There was a tendency of linear increase in nitrogen and phosphorus removal time with increasing concentration of swine wastewater. Under the optimized culture condition, total nitrogen and total phosphorus were effectively removed to 95.3% and 96.0%, respectively, in 25% swine wastewater after 4-day incubation.

도로변에 인접한 경작지 토양 및 작물체중의 연오염(鉛汚染) : 제1보 경작지 토양중의 연오염(鉛汚染) 및 화학적 형태 Ⅰ. Pollution of Pb in paddy field soil and its chemical forms

김장억,이석준 한국환경농학회 1991 한국환경농학회지 Vol.10 No.1

自動車 排氣가수에 包含된 鉛化合物의 排出에 의하여 汚染이 되었을 것으로 豫想되는 大邱直轄市를 中心으로 慶尙北道 內의 高速道路 및 主要國道邊에 隣接한 18個 地點을 選定하여 道路로부터 距離에 따른 畓土壤에서 鉛汚染度 및 土壤中 鉛化合物을 KNO₃浸出性(置換態), Na₄P₂O_7 浸出性(有機態), EDTA 浸出性(炭酸態) NHO₃ 浸出性(黃酸態), 非浸出性(不溶態)로 形態別 分布를 調査한 結果는 다음과 같다. 1. 畓土壤中 平均 鉛含量은 淸道에서 30.0 ppm으로 가장 높았고 高靈에서 14.8 ppm으로 가장 낮았으며 平均 21.9 ppm 으로 나타났으며, 交通量과 關聯하여 土壤中 鉛含量은 差異를 發見할 수 없었으나 道路의 開通年度에 따라 약간의 差異를 나타났다. 2. 道路로부터의 距離에 따른 鉛含量은 10m 以內에서 22.2 ppm, 10∼30m에서 22.1 ppm, 30∼50m에서 22.2 ppm, 50m 以上 地域에서 21.3 ppm으로 道路로부터 距離에 따른 畓土壤中의 鉛汚染度는 크게 差異 나지 않았다. 3. 土壤中 鉛化合物을 形態別로 分別 抽出한 結果 여러 가지 土壤特性에 따라 差異가 深하나 18個 地點에서 平均한 鉛化合物의 化學的 形態別 分布比는 有機態 33.6%, 炭酸態 29.8%, 黃酸態 21.5%, 置換態 8.6% 및 不溶態 6.7%의 順으로 나타났다. 4. 土壤中 有機物含量이 增加할수록 有機態 鉛의 分布比는 增加하는 傾向을 나타낸 반면 置換態 鉛의 分布比는 減少하는 傾向을 나타내었다. 5. CEC가 增加할수록 有機態의 鉛의 分布比는 增加하는 傾向을 나타낸 반면 置換態 鉛의 分布比는 減少하는 傾向을 나타내었으며, CEC에 依한 土壤中 鉛化合物의 化學的 形態別 分布比는 有機物에 依한 分布比와 비슷한 傾向을 나타내었다. The object of this stduy was to investigate the pollution of Pb in paddy field soil with different distance from roadside and to find out the relationship between the ratio of chemical fractions of total Pb and soil characteristics. Lead from automobiles is exhausted as particulates composed primarily of halide compounds (PbBrCl, PbBr₂, PbCl₂). The samples of soil were collected directly from the paddy fields with different distance from the roadside of highway and expressway which are located in Kyungpook province. A sequential extraction procedure was used to fractionate Pb in paddy field soil into the disignated forms of water soluable, exchangeable, organically bounded, carbonate, sulfide, and residual Pb. Results obtained are summerized as follows. 1. The content of Pb in paddy field soil was the highest in Chungdo, 30.0 ppm, the lowest in Koryung, 14.8 ppm, and the total average content was 21.9 ppm. The effect of traffic volume was not clear, but a slight difference according to the order of opened year of roads was showed. 2. The effect of distance from roadside was not clear. The content of Pb in paddy field soil with different distance from roadside was 22.2 ppm within l0m, 22.1 ppm in 10∼30m, 22.2 ppm in 30∼50m. and 21.3 ppm beyond 50m. 3. The distribution of Pb fractions in soil showed a wide difference depending on soil properties. The average content of exchangeable. organically bounded, carbonate, sulfide, and residual Pb was 8.6%, 33.6%, 29.8%, 21.5%, and 6.7%, of total Pb in the soil, respectively. 4. The content of organically bounded Pb in soil showed highly positive correlation with organic matter and CEC, while the content of exchangeable Pb was highly negative correlation. 5. With higher soil organic matter and CEC, organically bounded Pb fraction tend to be higher but exchangeable Pb fraction tend to be lower. Other forms of Pb showed no difference with soil organic matter contend and CEC. The distribution of Pb fraction related to CEC showed similiar tendency with that of organic matter content.

미세조류에 의한 지구온난화기체의 고정화

오희복,김정석,이석준 한국환경생물학회 1998 환경생물 : 환경생물학회지 Vol.16 No.4

전세계적으로 문제시되고 있는 과도하게 방출되는 CO_2에 의한 지구온난화의 문제를 생물학적으로 해결하기 위한 방법으로서 미세조류의 대량배양은 대기중 CO_2의 농도를 감소시키고, 배지로서 축산폐수를 이용할 경우에 질소와 인과 같은 영양염류의 농도를 현저히 감소시켜 당면한 호소 부영양화문제의 해결에도 일익을 담당하는 등 지구환경보존의 측면에서도 유망한 기술분야로 판단된다. 또한 배양된 조류의 생물량은 유용물질로 개발하여 이용할 수 있다. 식품이나 사료로서 미세조류의 영양적 가치는 많은 연구를 통하여 입증되었으며, 조류추출물의 상업적 가치도 인정되고 있다. 지금까지 알려진 수많은 조류종 중에서 오직 일부를 대상으로 경제적 가치 여부가 조사되었을 따름이다. 현재 조류로부터 물질생산은 작물로부터의 생산에 비하여 경제성을 비교하기는 어렵지만 조류배양의 비용이 점차 낮아지고 있으며, 이것을 수요로 하는 시장이 점차 확대되고 있다. 조류배양의 가장 큰 장점은 작물생산에 적합하지 않은 염분농도가 높거나 강한 알카리 등의 극한 환경에서도 성장 가능하다는 점이다. 또한 근래에 급속히 발전한 미생물 탐색, 미생물 배양, 유전공학적 조류주 개량, 유용물질 탐색 등 생명공학기술의 기반·핵심기술을 이용한다면 앞으로 미세조류를 이용한 천연색소의 생산, 유용 의약품 원료 생산, 건강식품 생산 등의 분야에도 직결되어 생물산업 분야의 큰 발전이 기대된다. 끝으로, CO_2의 대량배출처에서 미세조류의 대량배양에 의하여 CO_2 저감을 실현함으로서 환경분야의 범 지구적 관심사인 지구온난화 방지에 기여함과 동시에 배양된 미세조류로부터 고부가 생물자원을 생산함으로서 CO_2의 생물학적 고정화 기술의 경쟁력 강화 및 생물산업의 발전을 촉진할 수 있다고 판단된다. Carbon dioxide (CO_2) is viewed as the most potential agent among global warming gases. Photosynthetic CO_2 fixation with microalgae has emerged as a promising technology to help reduce emissions from fossil fuel-fired plants. Microalgae are of particular interest because of their rapid growth rates and tolerance to varying environmental conditions. Many researches have been focused on the followings: 1) the screening of microalgae growing under high temperature, high CO_2 and acidic condition, 2) the design of new reactor systems which are capable of fixing CO_2 effectively including light collecting system, and 3) the development of microalgal strains by using genetic engineering technology. The utilization of the biomass of microalgae as useful substances is also considered very important to be a cost-effective method of reducing the amount of CO_2 from fossil fuel-fired plants. In near future, it seems that the biological CO_2 fixation with microalgae will be applied and practiced in the industrial field and then resulted in both the reduction of CO_2 emission and the production of valuable substances.