• Food Industry And Nutrition
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• v.2 no.1
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• pp.7-9
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• 1997

내열설 미생물, Thermus caldophilus CK24에 대한 탄수화물 생합성을 연구하는 과정에서 다양한 탄수화물 관련효소를 탐색하고 그레 대한 생화학적 및 분자생물학적 연구를 수행하고 있다. 일차로 내열성 미생물내 1) 당핵산염 합성효소와 당전이 효소, 2) 탄수화물 대사효소. 3)탄수화물 분해 및 전환효소의 존재를 HPLC/Bio-LC분석을 통하여 확인하고 이들에 대한 연구를 진행하고 있다. 본 연구발표에서는 포도당을 과당으로 전환하는 이성화효소(xylose isomerase), 그리고 맥아당을 트레할로스로 전환하는 트레할로스 합성효소(trehalose synthase)를 소개하고저 한다. 이성화효소는 이미 산업적 과당 생산에서 대규모적으로 사용되고 있는 식품산업효소이다. 본 연구에서는 Thermus caldophilus GK24, Thermus thermophilus HB8, Thermus flavus AT62 3종의 내열성 미생물에 대한 이성화효소 유전자를 클로닝 하고, 각 재조합하고 이성화효소를 대량생산하였다. 이 내열성 이성화효소는 최적 반응 온도가 8$0^{\circ}C$이고, 포도당을 과당으로 전환하는 수유른 55%이었다. 이러한 과당전환률은 이미 산업적으로 사용되고 있는 이성화효소의 과당전환률(43%)보다 훨씬 높은 것으로 과당 생산공정의 단순화의 생산성 향상에 결정적인 요인이라 할 수 있다. 한편 본 이성화효소의 산업적 특성을 증대하기 위하여 구조-기능관계 연구를 착수하였다. 우선 내열성 이산화 효소의 입체 구조를 결정하였고, 구조조정에 따른 기능적 특성을 조사하기 위하여 특정 위치의 선택적 변이 연구를 진행하고 있다. 끝으로 포도당 전이 효소를 추적하던 과정에서 맥아당을 트레할로스로 전환하는 새로운 효소를 Thermus caldo-philus GK24에서 발견하였다. 그 트레할로스 합성효소는 분자량이 약 110kDa이고 최적 반응온도가 75$^{\circ}C$이면, 조효소없이 맥아당을 트레할로스로 80%이상 전환해 주는 가역효소이었다. 본 연구에서는 효소반응의 조건과 특성을 조사하였고, 효소 아미노-밀단의 서열결정정보를 통하여 효소의 유전자를 클로닝 하고 그 유전자의 구조와 발현연구를 진행하고 있다.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.238-238
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• 1994

녹용의 알려진 약리작용 중에서 녹용의 상용량 투여에 의해 나타나는 것은 강장작용과 창상치유작용이므로 이러한 작용이 진정한 녹용의 효능으로 간주될 수 있으나 이러한 작용을 생물활성 검색법으로 활용하여 녹용의 유효성분을 검색하기에는 재현성과 간편성이라는 면에서 어려운 점이 없지 않다. 녹용의 효능이 생체의 질소평형이 음의 방향으로 진행되었을 때 이를 정상 수준으로 복귀시키는 생물활성에 기인할 것이라는 가정하에서, 실험적으로 생체내 질소편형을 음의 방향으로 전환시킬 수 있는 급성기 반응 (acute-phase responce)을 야기시킨 다음 녹용의 효과를 연구하였다. 흰쥐의 급성기 반응물질 (acute-phase reactants, APR) 중에 서 positive ARP인 ceruloplasmin, $\alpha$-cysteine pretense inhibitor(= T-kininogen)는 비색정량법으로 간편하게 측정될수 있음에 착안하여 이 단백질을 지표로 삼아 turpentine oil로 급성 염증을 유발시킨 전후에 녹용 물추출액의 효과를 측정하였다. 그 결과 녹용의 상용량에서 괴소 4일 이상 경구투여한 흰쥐에서 급성 염증유발 후 4일에서 ceruloplasmin 의 혈중농도가 현저히 감소함을 알수 있었다. 급성 염증 유발후 녹용의 경구투여한 실험에서는 적어도 녹용 상용량의 3배 이상을 투여하여야 만 ceruloplasmin 혈중 농도를 감소시킬 수 있었다. 본 연구에서 수립된 생물활성을 이용하여 녹용의 유효 성분을 단리할 계획이다.

• Journal of Plant Biology
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• v.35 no.4
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• pp.365-370
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• 1992

다양한 배양조건을 이용하여 당근 모상근의 생장과 carotenoid 함량을 조사하였으며, 이러한 배양조건을 airlift형생물 반응기에 적용하여 모상근의 다량 배양을 시도하였다. 고형배지 배양에서의 carotenoid 함량은 광조건에서 배양된 모상근이 암조건에서 배양된 모상근보다 10배 정도 증가하였으며, 모상근의 생장은 광조건이 암조건보다 낮은 수준이었다. 공기 유출기 배양에서는 모상근이 광조건과 암조건 모두에서 매우 빠르게 생장하여 생줄량의 차이가 없었으며, 모상근의 생장과 carotenoid 함량에 있어서 빛의 조사는 모상근의 전체적인 생중량을 증가시키기 보다는 배양 후기 생장단계에서 carotenoid 함량을 증가시켜서 암조건에 비하여 10배 정도의 carotenoid(11 $\mu\textrm{g}$/g)를 생산하였다. 모상근을 생물 반응기에서 배양하였을 때, 모상근이 지수함수적으로 빠르게 생장하여 배양 25일 동안에 처음 무게의 11배를 나타내었다. 그리고 pH는 처음 5.5에서 5일후 4.9로 떨어지다가 그 이후에 일정한 수준(pH 4.8)을 보여주었으며, 광주기와 광도을 조절하지 않은 상태에서의 carotenoid 함량은 2$\mu\textrm{g}$/g으로 나타났다.

• Journal of Plant Biotechnology
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• v.33 no.3
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• pp.185-194
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• 2006

Chloroplasts are believed to be descended from certain cyanobacteria, which were taken up by phagocytosis into a host cell and lived there in a symbiotic relationship. In contrast to the current static concept on the chloroplast genome, its dynamism has been recently demonstrated: the chloroplast genome is active in intramolecular homolgous recombination, producing subgenomic circles when it obtains homolgous sequences via genetic transformation. Chloroplast tranformation in higher plants provides many advantages over nuclear transformation that include higher expression levels of transgenes, polycistronic expression of transgenes, and maternal transmission of transgenes. Tobacco has been used as a model for chloroplast genetic transformation. However, it is recently possible to transform the chloroplasts of other major food and economic crops including rice, soybean, and cotton. Chloroplast-transformed crops will be able to replace bioreactors using microorganisms for production of value-added proteins in future.

• Korean Journal of Medicinal Crop Science
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• v.8 no.1
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• pp.9-13
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• 2000

To determine the proper carbon and nitrogen sources and their proper levels for mass micro propagation of Scrophularia buergeriana Miquel, tonic and curing cough experiment were applied and a method for mass cultivation by using bioreactors (2.5 L) was expinined. Proper ratio of $NH_4NO_3\;:\;$KNO_3$ was 413 mg/L : 1900 mg/L for multiple shoot production. Sucrose was more effective than glucose or fractose as carbon source and 3% concentration was good for shoot formation. Total nitrogen was not detected after six weeks both in 500 ml flask and bioreactor culture. Sucrose was decreased sharply after two weeks and there was no sucrose left after three weeks both in 500 ml flask and bioreactor culture. The stirrer in bioreactor caused shear stress to shoots severely. The sphere type bioreactor was better than the cylinder type and removal of inner loop in sphere type was more effective to avoid shear stress.

• Research in Plant Disease
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• v.18 no.3
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• pp.186-193
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• 2012

Biotransformation is an eco-friendly and efficient method for enhancing the bioavailability of biopesticide. To increase the antifungal activity of the crude extract of Cynanchum wilfordii roots against barely powdery mildew, we performed biotransformation of wilfoside C1G using ${\beta}$-glucosidase (cellobiase from Aspergillus niger). The mixture (G sample) of partially purified wilfoside C1G and cynauricuoside A (K1G) was treated with ${\beta}$-glucosidase to remove a glucopyranosyl moiety. The enzyme completely converted C1G to C1N and K1G to K1N. Optimal conditions for enzymatic biotransformation of G sample were determined to be 10% ethanol, 1,555 ${\mu}U$ ${\beta}$-glucosidase/ml, pH 5, and $45^{\circ}C$. In in vivo experiment, the G sample transformed by ${\beta}$-glucosidase showed stronger antifungal activity against barley powdery mildew than the non-treated G sample. These results suggest that ${\beta}$-glucosidase biotransformation can be applied to increase the antifungal activity of the crude extract of C. wilfordii roots against powdery mildews.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.12 no.10
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• pp.4434-4442
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• 2011

Reactive oxygen species (ROS) are reactive and potentially harmful to cells, causing oxidation of lipids, proteins, and DNA. In humans, the deleterious effects of ROS have been linked with aging, carcinogenesis, and atherosclerosis. In order to investigate an antioxidant activity of wood vinegar by bioconversion, we preferentially analyzed the total acidity, acetic acid, pH, and contents of total polyphenols and flavonoids, respectively. Also, we evaluated the scavenging abilities on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, superoxide anion radicals, hydrogen peroxide radicals, and nitric oxide radicals. The total acidity and amount of acetic acid of wood vinegar after bioconversion were lower than those of wood vinegar before bioconversion, but the pH was higher than that of wood vinegar before bioconversion. The contents of total polyphenols and flavonoids of wood vinegar after bioconversion were 11.17 mg/$m{\ell}$ and 0.42 mg/$m{\ell}$, respectively. The $SC_{50}$ values were in order of superoxide anion radical scavenging activity < DPPH radical scavenging activity < hydrogen peroxide radical scavenging activity < nitric oxide radical scavenging activity. Therefore, these results suggest that wood vinegar by bioconversion can be useful as primary antioxidants for medicines and cosmetics.

• KSBB Journal
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• v.6 no.3
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• pp.309-319
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• 1991

A continuous stirred tank membrane reactor(CSTMR ) was developed and optimized for the production of cod skin gelatin hydrolyzates using endo-protease Alcalase. A experimental design methodology was used to optimize the four performance variables: enzyme concentration, substrate concentration, permeate flux and reactor volume. All four variables studied had an effect on substrate conversion, with enzyme and substrate concentrations being predominant. Conversion increased with the increase in enzyme concentration, with the decrease in substrate concentration, at high volumes and low flux. A strong interaction was observed between enzyme and substrate concentrations and smaller interactions between enzyme and flux and substrate and flux. The optimum operating conditions for the CSTMR process for an initial substrate concentration for 10% were $50^{\circ}C$, pH 8, flux 7.3ml/min, residence time 82 min, and Alcalase to substrate ratio 0.02(w/w). A gradual decay in reactor activity during 8 hrs was 2.1% conversion/hr. Enzyme leakage through the 10, 000 MWCO membrane was 16% at $50^{\circ}C$ and 12% at $35^{\circ}C$, 6hrs. However, there was no apparent correlation between enayme leakage and substrate conversion. The Km value for the CSTMR was 20 times higher than the batch reactor. The productivity(expressed as mg product/mg enzyme) of the CSTMR was more than six fold higher than the batch at $50^{\circ}C$. The hydrolyzate was non-bitter.

• Journal of the Korean Institute of Intelligent Systems
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• v.15 no.6
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• pp.682-687
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• 2005

Behavioral sequences of the medaka(Oryzias latipes) were investigated through an image system in response to medaka treated with the insecticide and medaka not treated with the insecticide, diazinon(0.1 mg/1). After much observation, behavioral patterns could be divided into 4 patterns: active smooth, active shaking, inactive smooth, and inactive shaking. These patterns were analyzed by 5 features: speed ratio, x and y axes projection, FFT to angle transition, fractal dimension, and center of mass. Each pattern was classified using decision tree. It provide a natural way to incorporate prior knowledge from human experts in fish behavior, The main focus of this study was to determine whether the decision tree could be useful in interpreting and classifying behavior patterns of the animal.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.216-219
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• 2000

Pyrolysis products of woody biomass were consistedofvarious linear hydrocarbon, aromatics or condensed cyclic compounds. In order to obtain biomass pyrolysis products, more equipments and time were needed. But solvolysis of woody biomass with acetone easily obtained decomposition products and enhanced conversion rate(18.72%, max.) from woody biomass than pyrolysis of woody biomass. Beacause solvolysis with acetone improved conversion rate (26.64%, max.) of lignin. the whole conversion rate was improved. But above $300^{\circ}C$, lignin showed lower conversion rate, so the whole conversion rate decreased. Solvolysis products of woody biomass with acetone were same as pyrolysis products. Above $400^{\circ}C$, methoxy phenols were completely disappeared.