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구름버섯(Coriolus versicolor IFO 30388)에 의한 Poly R-478 염료의 탈색
윤경하,Yoon, Kyung-Ha 한국미생물학회 1994 미생물학회지 Vol.32 No.3
구름버섯에 의한 Poly R-478염료의 탈색에 미치는 탄소원과 질소원의 영향을 조사하였다. 2.0% glucose, 0.04% ammonium tartrate, 2% $KH_2PO_4$, 0.5% $MgSO_4{\cdot}7H_2O$, 0.1% $CaCl_2{\cdot}2H_2O$, 0.002% thiamine-HCI, 10 mM 2,2-dimethylsuccinate(sodium)와 0.02% Poly R-478 염료 등으로 조성된 질소제한배지에서 균주를 $28^{\circ}C$에서 10일간 정치배양 했을때 염료의 탈색율은 87.2%로 나타났다. 염료의 탈색은 배지내에 질소원이 존재할 때에 일어났으며 탈색율은 배지내에 질소원이 고갈된 직후에 급격히 증가하였다. Effects of nitrogen and carbon sources on the decolorization rate of poly R-478 dye by a white rot basidiomycete Coriorus versicolor IFO 30388 were examined. The fungus exhibited 87.2% of decolorization rate when it was cultured in the state of stationary in a nitrogen-limited medium (pH 4.5) which contained 2.0% glucose, 0.04% ammonium tartrate, 0.02% poly R-478 dye, 2% $KH_2PO_4$, 0.5% $MgSO_4{\cdot}7H_2O$, 0.1% $CaCl_2{\cdot}2H_2O$, 0.002% thiamine-HCl and 10 mM 2,2 dimethylsuccinate (sodium) at $28^{\circ}C$ for 10 days. Decolorization of the dye occurred in the presence of nitrogen source in the medium and decolorization rate increased rapidly after depletion of $NH_4^+$ from the medium.
폐광지역에서 분리한 Benzoate 분해세균 Pseudomonas sp. NEQ-1에서 정제된 Catechol 1,2-Dioxygenase의 특성
주정수,윤경하,Joo Jung-Soo,Yoon Kyung-Ha 한국미생물학회 2004 미생물학회지 Vol.40 No.4
Quinoline (2,3-benzopyridine)을 유일한 탄소원과 질소원, 그리고 에너지원으로 이용하는Pseudomonas sp. NEQ-1을 실험 균주로 사용하였으며, 균주로부터 catechol 1,2-dioxygenase (C1,2O)를 유도하기 위하여 탄소원으로 benzoate를사 용하였다. C1,2O의 효소학적 특징을 조사하기 위하여 benzoate에서 배양한 Pseudomonas sp. NFQ-1을 초음파 분쇄기로 파쇄하고, ammonium sulfate침전과 gel permeation chromatography및 Source 15Q의 과정을 실시하여 C1,2O를 분리 및 정제하였다. 정제된 C1,2O의 특이활성(specific activity)은 14.21 unit/mg으로 나타났으며, SDS-PAGE에 의해 조사된 C1,2O의 분자량은 약 33 kDa이었다. Cl,2O는 catechol과 4-methylcatechol 및 3-methylcatechol에 대해서 효소활성을 나타내는 것으로 확인되었다. C1,2O의 Km은 38.54 ${\mu}M$로 측정되었고, Vmax는 $25.10\;{\mu}mol{\cdot}min^{-1}{\cdot}mg^{-1}$으로 나타났다. C1,2O는 $30^{\circ}C$와 pH 8.5에서 최적활성을 나타내는 것으로 조사되었으며, $Ag^+,\;Hg^+,\;Ca^{2+}$,그리고 $Cu^{2+}$는 C1,2O의 활성을 억제하였다. 분석되어진 N-말단 아미노산 서열은 ^1TVKISQSASIQKFFEEA^{17}$이었으며, Pseudomonas aeruginosa PA01과 $82\%$로 가장 높은 유사성을 보였고 Pseudomonas arvilla C-1와는 $71\%,$ Pseudomonas putida KT2440과는 $59\%,$ 그리고 Pseudomonas sp. CA10과는 $53\%$의 상동성이 각각 존재하는 것으로 확인하였다. Our previous research has demonstrated that the bacterium, Pseudomonas sp. NFQ-l capable of utilizing quinoline (2,3-benzopyridine) as the sole source of carbon, nitrogen, and energy was isolated and characterized [Yoon et ai. (2003) Kor. J. Biotechnol. Bioeng. 18(3):174-179]. In this study, we have found that Pseudomonas sp. NFQ-l could degrade quinoline as well as benzoate, and extended this work to characterize the catechol 1,2dioxygenase (C1,2O) purified from the bacterium cultured in benzoate media. Initially, C1,2O has been purified by ammonium sulfate precipitation, gel permeation chromatography, and Source 15Q. After Source 15Q, purification fold was increased to approximately 14.21 unit/mg. Molecular weight of C1,2O was about 33 kDa. Physicochemical characteristics (e.g., substrate specificity, Km, Vmax, pH, temperature and effect of inhibitors) of purified C1,2O were examined. C1,2O demonstrated the activity for catechol, 4-methylcatechol and 3-methylcatechol as a substrate, respectively. The Km and Vmax value of C1,2O for catechol was 38.54 ${\mu}M$ and $25.10\;{\mu}mol{\cdot}min^{-1}{\cdot}mg^{-1}.$ The optimal temperature of C1,2O was $30^{\circ}C$ and the optimal pH was approximately 8.5. Metal ions such as $Ag^+,\;Hg^+,\;Ca^{2+},\;and\;Cu^{2+}$ show the inhibitory effect on the activity of C1,2O. N-terminal amino sequence of C1,2O was analyzed as ^1TVKISQSASIQKFFEEA^{17}.$ In this work, we found that the amino acid sequence of NFQ-l showed the sequence homology of 82, 71, 59 and $53\%$ compared with C1,2O from Pseudomonas aeruginosa PA0l, Pseudomonas arvilla C-1., P. putida KT2440 and Pseudomonas sp. CA10, respectively.
주걱송편버섯(Pycnoporus cinnabarinus SCH-3)의 Laccase에 의한 Acridine 산화
이현수,한만덕,윤경하,Lee, Hyoun-Su,Han, Man-Deuk,Yoon, Kyung-Ha 한국미생물학회 2008 미생물학회지 Vol.44 No.2
Acridine은 fungal laccase의 기질이 아님에도 불구하고 acridine을 Pycnoporus cinnabarinus SCH-3 배양액에 첨가했을 때 acridone으로 산화되었다. P. cinnabarinus SCH-3균주는 배양 중에 다량의 laccase와 3-hydroxyanthranilic acid (3-HAA)와 cinnabarinic acid (CA)를 생성했다. 정제된 laccase와 acridine을 완충용액에서 직접 반응시켰을 때 acridine은 변화되지 않았다. 그러나 laccase의 기질인 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS)나 3-HAA를 laccase와 acridine 혼합액에 첨가했을 경우에는 acridine이 acridone으로 산화되었다. 특히 ABTS 첨가구는 3-HAA 첨가구보다 acridine 산화율이 2배 이상 높았다. 한편 정제된 laccase와 3-HAA를 완충액에서 반응시켰을 때 3-HAA는 CA로 전환되었다. 이와 같은 실험결과들은 P. cinnabarinus SCH-3의 laccase가 배양중에 생산된 3-HAA를 매개체로 사용하여 acridine을 acridone으로 산화하고 CA는 laccase에 의하여 3-HAA로부터 합성됨을 나타낸다. Acridine was not a substrate for fungal laccase but it was oxidized to acridone in the culture medium of P. cinnabarinus SCH-3. During the cultivation of P. cinnabarinus SCH-3, Laccase was the predominant extracellular phenoloxidase, and 3-hydroxyanthranilic acid (3-HAA) was produced in the early culture. Cinnabarinic acid (CA) was observed to accumulate in the culture medium. When P. cinnabarinus was grown in the culture medium containing acridine, acridine was oxidized to acridone. But when the laccase purified from the culture medium of P. cinnabarinus directly reacted with acridine in sodium tartrate buffer (pH 3.0), The oxidation of acridine did not happen. In contrast, when 3-HAA was added to the buffer that was mixed with laccase and acridine, the acridine was oxidized to acridone. While in vitro studies, the CA was formed from 3-HAA in the presence of purified laccase. The results suggest that the acridine should be oxidized to the acridone through the mediation of 3-HAA by the laccase in the culture medium of P. cinnabarinus SCH-3.
기온 변화 특성을 반영한 하절기 전력수요예측에 관한 연구
구본희(Bon-Hui Ku),윤경하(Kyoung-Ha Yoon),차준민(Jun-Min Cha),송경빈(Kyung-Bin Song),백웅기(Ung-Ki Baek) 대한전기학회 2010 대한전기학회 학술대회 논문집 Vol.2010 No.11
This paper proposes a short-term load forecasting algorithm using the sensitivity of temperature in a summer season. The sensitivity of temperature is used to improve the accuracy of the load forecasting through the relation of daily load and daily high temperature. Case studies were applied to load forecasting for summer of 2009, and the results shows its appropriate accuracy.
영지의 ${\beta}$-glucan성 다당류에 의해 활성화된 흰쥐 간내 Kupffer 세포의 NO, TNF-${\alpha}$ 및 TGF-${\beta}$ 형성
한만덕,이준우,정훈,김용석,나수정,윤경하,Han, Man-Deuk,Lee, June-Woo,Jeong, Hoon,Kim, Yong-Seok,Ra, Su-Jung,Yoon, Kyung-Ha 한국미생물 · 생명공학회 1999 한국미생물·생명공학회지 Vol.27 No.1
Ganoderan (GAN), an immunomodulating ${\beta}$-glucan from mushroom Ganoderma lucidum, was evaluated for its ability to induce formation of nitric oxide (NO), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) and transforming growth factor (TGF-${\beta}$) from rat Kupffer cell in vitro. Hepatic macrophages activated by GAN significantly elevated concentration of NO and TNF-${\alpha}$ in cultured medium, but not significantly elevated that of TGF-${\beta}$. GAN-activated Kupffer cells secrete 14.9${\mu}$M (p<0.01) of NO and 2619.5${\rho}$g/ml (p<0.01) of TNF-${\alpha}$after 36hr of incubation at 37$^{\circ}C$. The results revealed that GAN enhanced 4-fold production of NO and 19 fold formation of TNF-${\alpha}$ compared to the control. The proliferation of GAN-activated Kupffer cells was inhibited as compared with its negative control. Comparing the activity among glucans derived from microorganisms, highly branched zymosan, glucomannan from Saccharomyces cerevisiae, significantly increased TNF-${\alpha}$ and NO production. These results indicate that the ${\beta}$-glucan from G. lucidum activates rat Kupffer cell and secretes NO and TNF-${\alpha}$. It also suggest that rat Kupffer cell posses certain receptor for ${\beta}$-anomeric glucan.