In this presentation, we will discuss several recent achievements developed in my laboratory for microarray-based diagnosis of human genetic mutations including HNF-1 and BRCA1 mutations. To determine the presence of the genetic mutations in a human sample, we prepared allele-specific oligonucleotide chips from selected mutation sites and generated target probes using a tow-step method for Cy-3 DNA $samples^{1)}$ or in vitro transcription of promoter-tagged PCR products for Cy-3 RNA $samples^{2)}$. Hybridization of the target probes to the chips successfully identified all of the genotypes for the tested sites. For more reliable diagnosis, we also employed single base extension (SBE) reaction and zip-code microarray technique for our strategy. Particularly we developed an efficient PNA zip-code microarray for the detection of $HNF-1{\alpha}$ $mutations^{3)}$. Using multiplex SBE reactions and zip-code strategy, we were able to correctly diagnose several mutation sites in exon 2 of $HNF-1{\alpha}$ with a wild-type and mutant including a MODY3 patient. These works represent successful applications of DNA microarray technology for the diagnosis of human genetic mutations.

Photoinduced electron transport process in nature such as photoelectric conversion and long-range electron transfer in photosynthetic organisms are known to occur not only very efficiently but also unidirectionally through the functional groups of biomolecules. The basic principles in the development of new functional devices can be inspired from the biological systems such as molecular recognition, electron transfer chain, or photosynthetic reaction center. By mimicking the organization of the biological system, molecular electronic devices can be realized $artificially^{1)}$. The nano-fabrication technology of biomolecules was applied to the development of nano-protein chip for simultaneously analyzing many kinds of proteins as a rapid tool for proteome research. The results showed that the self-assembled protein layer had an influence on the sensitivity of the fabricated bio-surface to the target molecules, which would give us a way to fabricate the nano-protein chip with high sensitivity. The results implicate that the biosurface fabrication using self-assembled protein molecules could be successfully applied to the construction of nanoscale bio-photodiode and nano-protein chip based on electrical detection.

This paper reports a novel magnetic force-based microfluidic immunoassay using microbeads and magnetic nanoparticles. The magnetic force-based immunoassay was devised first and successfully applied to detect the rabbit IgG as the model analyte of microfluidic sandwich immunoassay. The microchannels were fabricated by poly(dimethysiloxane) (PDMS) molding processes and bonded on a slide glass by plasma treatment. At the part of the inlet, sample solution was hydrodynamically focused. The focused microbeads of sample solution were flowed through the 150 ${\mu}m$ width channel of outlet. However, when the microbeads are conjugated with the superparamagnetic nanoparticles under the applied magnetic fields, they will switch their flow path and flow through the 95 ${\mu}m$ width channel of outlet. The movements of microbeads conjugated with magnetic nanoparticles were demonstrated by magnetic field $gradients.^{1)}$ High magnetic field gradients using micro electromagnets could be applied to this detection method for high sensitivity and lower detection limit. In addition, the multiplexed $immunoassay^{2)}$ using an encoded microbead which is immobilized with a certain antibody could be possible using this detection principle.

Neoagaro-oligosaccharides are produced only by enzymatic degradation of agarose by ${\beta}-agarase.^{1)}$ Neoagaro-oligosaccharides inhibit the growth of bacteria, slow the rate of degradation of starch, are used as low-calorie additives to improve food quality, and have macrophage-stimulating activity. Furthermore, neoagarobiose is a rare reagent that has both moisturizing effect on skin and whitening effect on melanoma $cells.^{2)}$ An agar-degrading marine bacterium was isolated from the sea water at the northeast coast in Cheju island, Korea. The strain was gram negative, aerobic, and motile rod. The 16S rRNA of the strain had the closest match of 98% homology, with that from Agarivorans albus. On the basis of several phenotypic characters and a phylogenetic analysis, this strain was designated Agarivorans sp. JA-1. In solid agar plate, Agarivorans sp. JA-1 produced a diffusible agarase that caused agar softening around the colonies. Agarivorans sp. JA-1 was cultured for 36 hr in marine broth 2216 (Difco, USA) and the supernatant that containing an extracellular ${\beta}-agarase$ was prepared by centrifugation of culture media. The enzyme exhibited relatively strong activity at $40^{\circ}C$ and was stable up to $60^{\circ}C$. Using PCR primers derived from the ${\beta}-agarase$ gene of Vibrio sp., the gene encoding ${\beta}-agarase$ from Agarivorans sp. JA-1 was cloned and sequenced. The structural gene consists of 2931 bp encoding 976 amino acids with a predicted molecular weight of 107,360 Da. The deduced amino acid sequence showed 99% and 34% homology to $agaA^{2)}$ and $agaB^{2)}$ genes for ${\beta}-agarase$ from Vibrio sp., respectively. The expression plasmid for ${\beta}-agarase$ gene of Agarivorans sp. JA-1 is being constructed and the recombinant enzyme will be biochemically characterized.

Recombinant human $interferon-{\beta}-1a(rIFN-{\beta})$ is a single glycosylated protein (at N80, 1N) with anti-viral activity. However, present drugs have a relatively short serum half-life of $rIFN-{\beta}$, thus patients suffer from frequent $infections.^{1)}$ To improve its half-life, eight glycosylation analogs were prepared, which have additional N-linked glycosylation consensus sequences (N-X-T/S) within the $IFN-{\beta}$ molecule and/or at C-terminal. Each $rIFN-{\beta}$ analog was examined for the presence of additional N-linked glycosylation and the maintenance of anti-viral activity in CHO cells. The molecular weights of five analogs were not changed. However, two analogs, R27T within $rIFN-{\beta}$ (27 kDa, 2N) and GNITVNITV at C-terminal (29kDa, 2N), showed a clear increase in molecular weights, compared to native $rIFN-{\beta}$ (23 kDa, 1N). And another combined analog of R27T+GNITVNITV showed increased molecular weight (33 kDa, 3N). It was confimed that the molecular weight increment of analogs was caued by the N-linked glycosylation with the treatment of N-glycansae. In the case of anti-viral activity, the analog GNITVNITV showed a reduction in activity compared to native $IFN-{\beta}$, whereas the analogs R27T and R27T+GNITVNITV were found to have distinctly increased activities. Pharmacokinetic study in rats also disclosed that the analogs R27T and R27T+GNITVNITV had 2 3 fold increased serum half-life, respectively. In conclusion, the addition of N-linked glycosylation in $rIFN-{\beta}$ increased serum half-life, thereby its less frequent administration will be expected.

Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone consisting of non-covalently linked two subunits, the ${\alpha}$ and ${\beta}$ subunit. It has been used as a infertility drug for ovulation to mimic luteinizing hormone $(LH).^{1)}$ A stable cell line was established by transfection of Rc/CMV-i-dhfr-hCG, expression vector containing hCG ${\alpha}-$ and ${\beta}-genes$, into dihydrofolate reductase-deficient CHO cells and subesquent methotrexate-mediated gene amplification. Anchorage-dependent CHO cells were adapted into a serum-free and/or animal component-free suspension medium through gradual serum weaning for the hCG production. The established cell line showed typical morphological characteristics and growth profile of CHO cells, and could produce FSH with passage-to-passage consistency. The high density perfusion culture of the CHO cells was carried out in Celligen Plus bioreactor equipped with a spin-filter as a internal cell retention device. The cell density reached up to $>1x10^{7}$ cells/ml in less than 7 days and a perfusion-control strategy based on cellular consumption rates of glucose was $established.^{2)}$ Biologically active recombinant hCG was purified by a series of chromatographic steps including anion exchange chromatography and hydrophobic interaction chromatography to homogeneity. The highly purified recombinant hCG was characterized for physicochemical, immunological and biological properties.

Large scale mammalian cell culture has become, over the past two decades, the preferred method to produce therapeutic monoclonal antibodies. In this presentation, I will introduce disposable bioreactor system and analyze key factors and points for consideration during mammalian cell culture process development. Example will be provided highlighting the selection of master cell, culture media and environmental factors based on productivity and product quality.

There are many different surface technologies currently applied for preparation of protein chips. However, it requires innovative surface chemistry for capture proteins to be immobilized on chip surface keeping their conformation and activity intact and their orientation right, while they bind tightly and densely in a given array spot. Proteogen has developed 'ProteoChip BP' coated with novel proprietary linker molecules $(ProLinker^{TM})$ for efficient and robust immobilizations of capture proteins by improving surface properties of molecular captures. It was demonstrated that $ProLinker^{TM}$ gave the best surface performance in preparation of protein microarray chip base plates among others currently available on the market. In particular, the $ProLinker^{TM}-based$ surface chemistry has demonstrated to provide excellent performance in preparation of 'Antibody Chip' for analysis of biomarkers as well as proteome expression profiles. The linker molecule has also shown to be well applicable for development of biosensors and micro-beads as well as protein microarray and nano-array. ProteoChip BP can be used either for preparation of high-density array by using a microarrayer or for preparation of 'Well-on-a-Chip' with low density array, which is better applicable for quantitative analysis of biomarkers or protein-protein interactions. The biomarker assay can be performed either by direct or sandwich methods of fluorescence immunoassay. Application of ProteoChip BP has been well demonstrated by the extensive studies of 1) tumor-marker assays, 2) new drug screening by using 'Integrin Chip' and 3) protein expression profile analysis. Some of experimental results will be presented.

Anaerobic digestion is one of the well-known methods for biological treatment handling of concentrated organic matter such as swine $wastewater.^{1)} The anaerobic digestion can reduce organic loading but also hydrolyze non-biodegradable organic $matter.^{2)}$ The feces from the scrapper-type barn are usually collected to make compost and the urine is discarded with swine-slurry wastewater by ocean-dumping or treated by biological methods. The lagoon, aerobic digestion, anaerobic digestion, SBR, $A^{2}/O$, and UCT have been applied for treating swine $wastewater.^{3)} In this study, as a result of the analysis of swine wastewater, the total and soluble chemical oxygen demand was 130g/L and 60g/L, respectively. And the volatile fatty acid as chemical oxygen demand equivalent was 45g/L, which was 75% of soluble chemical oxygen demand. Before everything else, ammonia nitrogen concentration was 6.5 g/L. From biochemical acidogenic potential test, it was concluded that the enhanced acidification process to manage swine waste should be operated in the ammonia nitrogen concentration of less than 1.2 g/L. In the result of seeding ratio experiments with artificial $wastewater^{4)}, the lag period of acidogens was taken the long time because of the inhibition by the $ammonia^{5)}$, however no difference of period by the seeding ratio was not shown. The Haldane-based biokinetics were also evaluated using a method of fourth order Runge-Kutta $approximation.^{6,7)}$ The nonlinear least squares (NLLS) method with a 95% confidence interval was also used. The ranges of maximum microbial growth rate, ${/mu_{max}}$, and half saturation coefficient, $K_{s}$, for acidogenesis of various seeding ratio with artificial wastewater were 6.1 ~ 12.6 $d^{-1}$ and 45,000 ~ 53,500 mg glucose/L, respectively. Also, the methanogenic microbial yield coefficient, Y, and microbial decay rate coefficient, $k_{d}$, and inhibition substrate concentration, $K_{si}$, for the reactors were determined to be 0.32 ~ 0.465 ${/mu}g$/mg glucose; 0.42 ~ 1.01 $d^{-1}$ and 51,500 ~ 55,600 mg glucose/L, respectively.

Leuconostoc citreum is one of the representative strains of Leuconostoc spp. that show fast growth rates in fermented vegetables. Sequential experimental designs including the Plackett-Burman design, fractional factorial design, steepest ascent analysis, central composite design and response surface methodology were introduced tooptimize and improve the medium for Leuconostoc citreum. Fifteen medium ingredients were examined and glucose (20 g/l), yeast extract (12.5 g/l), sodium acetate trihydrate (6.12 g/l), potassium phosphate (42.55 g/l) and dibasic ammonium citrate (4.12 g/l)were chosen as the best components to give a critical and positive effect for cell-growth. The biomass was increased to 2.79 g/l (169%), compared to the 1.65 g/l in MRS medium.

수궐음심포경을 전기 자극한 후 두 자극점내 경혈점에서 온도강하가 이루어짐으로 보아 수궐음심포경에 대한 자극은 열의 발산을 활발하게 한다고 생각되어진다. 50Hz에서 노궁 (PC8)과 극문 (PC4)사이 온도차이가 100Hz와 150Hz의 자극에 비하여 적게 나타남을 확인할 수 있었다. 따라서, 인체의 열에너지 발산정도가 주어지는 자극세기에 비례한다고 생각되어진다.

Doxorubicin is a highly-valuable anthracycline-family polyketide drug with a very potent anticancer activity, typically produced by a Gram-positive soil bacterium called Streptomyces peucetius. Thanks to the recent development of Streptomyces genomics-based technologies, the random mutagenesis approach for Streptomyces strain improvement has been switched toward the genomics-based technologies including the application of DNA microarray systems. In order to identify and characterize the genomics-driven potential target genes critical for doxorubincin overproduction, three different types of doxorubicin overproducing strains, a dnrI(doxorubicin-specific positive regulatory gene)-overexpressor, a doxA (gene involved in the conversion from daunorubicin to doxorubicin)-overexpressor, and a recursively-mutated industrial strain, were generated and examined their genomic transcription profiles using Streptomyces DNA microarray systems. The DNA microarray results revealed several potential target genes in S. peucetius genome, whose expressions were significantly either up- or down-regulated comparing with the wild-type strain. A systematic understanding of doxorubicin overproduction at the genomic level presented in this research should lead us a rational design of molecular genetic strain improvement strategy.

Aptamer is the single-stranded oligonucleotide which binds to various target molecules such as proteins, peptides, lipids and small organic molecules with high affinity and specificity. DNA aptamers specific for the $17{\beta}-estradiol$ were selected by SELEX (Systematic Evolution of Ligands by EXponential enrichment) process from a random DNA library. These DNA aptamers have a high affinity to $17{\beta}-estradiol$ as an endocrine disrupting chemical. Nanowell and $200{\mu}m$ gold electrode were used as substrate for DNA aptamer immobilization and electrochemical analysis. Especially, nanowell gold electrode was fabricated by e-beam lithography. The size of single nanowell is 130nm and 40,000 nanowells were deposited on one gold electrode. The immobilization method was based on the interaction between the biotinylated aptamer and streptavidin deposited on gold electrode previously. Immobilization procedure was optimized by surface plasma resonance (SPR) and electrochemical analysis. After the immobilization of DNA aptamer on streptavidin modified gold electrode, $17{\beta}-estradiol$ solution was treated on aptamer immobilized gold electrode. The current of gold electrode was decreased by the binding of $17{\beta}-estradiol$ to DNA aptamer immobilized on gold electrode. However, in negative control experiments of 1-aminoanthraquinone and 2-methoxynaphthalene, the current was rarely decreased. And more sensitive data was obtained from nanowell gold electrode comparing with $200{\mu}m$ gold electrode.

The Basidomycetes fungus Agaricus blazei Murill has been well known as a health food for the prevention of cancer, diabetes, hyperlipidemia, arteriosclerosis and chronic hepatitis. This study was concentrated to investigate the characteristics of crude protein-bound polysaccharides(PBP) compositions extracted from Agaricus blazei Murill. In order to produce crude polysaccharides, culture conditions were examined using YMK media. Total sugars and protein contents of PBP were detected by phenol-sulfuric acid method and Bradford -assay. Hexosamine was found to be involved in the linkage, N-linked and O-linked types. To identify helical conformation existence, wavelength was measured using Congo red after the treatment with alkali solution.

It has been known that the novel diterphenoids, hericenone and erinacine isolated from the fruiting body and cultured mycelia of Hericium erinaceus showed potent stimulating activity of nerve growth factor (NGF)-synthesis. To investigate the biological activities of extracts from fruiting body, cultured mycelium and cell-free broth of H. erinaceus, the activity experiments of antioxidation and AChE inhibition were carried out. Sephadex G-10 gel filtration followed by HPLC on a ${\mu}Bondapak$ $C_{18}$ column of EtOAc extract from cultured mycelium showed a biological activity.

Phospholipases C (PLCs) from B. cereus 318 and recombinant Pichia pastoris were immobilized on sol-gel coated glass beads. The pH and temperature on immobilized PLC activity were investigated. Operational and storage stability of the immobilized PLCs was measured by spectro- photometric assay. The PLCs immobilized on sol-gel coated glass beads were photographed by scanning electron microscope (SEM).

This studies were carried out to investigate optimal conditions for Lactic acid bacteria growth, which was grown in a batch fermenter. The optimal temperature was $30^{\circ}C$, optimal pH was 6.5 and agitation speed was 100rpm and didn't supply the air. Used media compositions were yeast extract 5g/L, peptone 10g/L, sugar 20g/L, beef extract 10g/L, tween 80 1ml/L, ammonium citrate 2g/L, sodium acetate 5g/L, magnesium sulfate 0.1g/L, manganese sulfate 0.05g/L, dipotassium phosphate 2g/L. These results would be useful for enhancing lactic acid bacteria concentration.

This study was concerned with the optimization of liquid culture conditions for mycelial growth and polysaccharide production and its physicochemical properties in Cordyceps militaris. The one factor at a time method was adopted to investigate the effects of medium composition, environmental factors and C/N ratio. Among the these varialbles, glucose 80g/L, yeast extract 10g/L, $MgSO_4{\cdot}7H_{2}O\;0.5g/L$, $KH_{2}PO_4\;0.5g/L$ were proved to be the most suitable carbon, nitrogen, and mineral sources, respectively. The optimal temperature, initial pH, working volume were identified to be $24^{\circ}C$, 7.0 and 100ml, respectively. Under the optimal conditions, the strategies in shake flask culture and 5L jar fermentor led to mycelial growth of 29.43 g/L, 28.88g/L and polysaccharide production of 2.53g/L, 6.38 g/L, respectively. Among the phisicochemical properties, relative concentrations(w/v) of total sugar, uronic acid, protein and hexoseamine were identified to be 74.07%, 1.13%, 0.91%, and 0.46%, respectively. The fraction of neutral and acidic polysaccharide were identified to be 81.9% and 18.1%, respectively.

본 연구에서 수행한 avermectin 생산을 위한 배지 최적화에서 기존의 총 avermectin의 함량 10 mg/L를 470 mg/L까지 증가시켰으며, 이 중 광범위 활성을 가지는 avermectin B1도 50%에 달했다. 최적화된 배지 조성은 50 g/L fructose, 30 g/L malt extract, 5 g/L casamino acid, 2.5 g/L PEG 3,350, and 1 g/L $K_{2}HPO_{4}$이다. $K_{2}HPO_{4}$와 fructose, malt extract가 avermectin 생합성에 크게 작용하는 것으로 추정된다.

정어리에서 분리한 5개의 균주를 이용하여 antiangiogenesis 효과가 가장 뛰어난 균주를 ${\lambda}-28$이라 명명하고 이를 대량 배양하여 단백질 추출법인 염석과 투석, 동결건조를 통해 시료를 제조하여 이를 SEC를 통해 fraction별로 분리한 후 SDS-PAGE와 antiangiogenesis, cytotoxcity test를 수행하여 ${\lambda}-28$번의 76 fraction에서 control군에 비해 82.7%의 높은 antiangiogenesis 효과를 보였고, 세포독성 실험에서도 낮은 독성을 보였다.

20 g/L peptone, 20 g/L dextrose, 10 g/L yeast extract에 100 mg/L zeocin을 첨가하여 동일하게 전배양 한 재조합 Pichia pastoris X-33/pBPT44를 각기 다른 탄소원이 든 배지에 배양하면서 12시간 간격으로 샘플을 채취하여 배양시간에 따른 세포성장, pH, 각 탄소원에 따른 PLC 생산량 등을 측정하였다.

In this study, we investigated the optimal medium composition for bacterial cellulose (BC) production by Gluconacetobacter sp. RKY5. Among the various kinds of carbon sources, glycerol was the most efficient as a sole carbon source and its optimal concentration for BC production was 15 g/L. The optimal concentration of yeast extract as a nitrogen source for BC production was found to be 8 g/L. $K_{2}HPO_{4}$ and acetic acid were selected respectively as a phosphate source and a secondary substrate, and both optimal concentrations were 3 g/L. The amount of produced BC was 4.59 g/L in a static culture and 6.5 g/L in a shaking culture condition with 150 rpm. These values were 2.1 and 2.7 times higher than those in a static (2.16 g/L) and a shaking (2.41 g/L) cultures using HS medium generally used for BC production.

We investigated the fermentative production of lactic acid from cheese whey and corn steep liquor as cheap raw materials using Lactobacillus sp. RKY2 to reduce the manufacturing cost of lactic acid. Lactic acid yields were obtained at more than 0.98 g/g from medium containing whey lactose. Lactic aid productivities and yields obtained from whey lactose were slightly higher than those obtained from pure lactose. The final concentration of lactic acid increased with increase, in whey lactose concentration, whereas the lactic acid productivity decreased probably due to substrate inhibition. The fermentation efficiencies were improved by addition of more corn steep liquor to the medium.

발효기에서의 균사체 생장과 세포외다당류의 생산을 위한 최적 배양조건을 조사하기 위하여 회분식 반응기를 이용하여 연구한 결과 최적 회전속도와 통기량은 각각 200 rpm과 1.5 vvm 었다. 최적 배양 환경하에서 5L발효기(working volume 2 L)에서 배양한 결과 각각 10.21 g/L 와 3.56 g/L의 mycelial growth와 exo- polysaccharide를 얻었다.

본 연구에서는 생물반응기에서 효율적으로 돼지 전염성 백신을 생산하기 위해 swine testicle cell을 host cell로 하는 백신 생산 시스템을 확립하였고, 이를 위한 최적 MOI, 최적 감염시기, 최적 수확시기 등 백신 생산 조건을 확립하였다. 또한 보다 안전한 백신 생산을 위해 통계적 방법에 의해 무 혈청 배지를 개발하였고 백신 생산 면에서의 가장 우수한 무 혈청 배지를 선정하였다.

Serum is a potential source of bacterial, mycoplasmal and viral contamination, and it has a possibility of the introduction of serum proteins, prion and pyrogens into the final vaccine product. For porcine Rotavirus vaccine production, it is necessary to develop serum free medium which do not cause those problems. A new serum free medium was developed for porcine Rotavirus vaccine based on DMEM, and the performance of developed serum free medium was evaluated in terms of Vero cell growth and Rotavirus vaccine production. The cell density, gown in serum free medium developed, was similar with that in serum supplemented medium. Also, it was higher than that in other commercially available serum free medium. The productivity of Rotavirus vaccine using serum free medium developed and optimum production strategies will be also discussed.

본 연구에서는 세포 사멸을 억제시켜 재조합 단백질의 생산을 향상시키기 위한 전략을 개발하기 위해, 우선 STR-G와 EGCG를 농도별로 첨가하여 세포 성장, 사멸 그리고 IDS 생산성에 미치는 영향을 조사하였다. Flow cytometry 분석 결과, STR-G에 의한 apoptotic cell percentage의 감소, STR-G와 EGCG에 의한 bel-2 expression level의 증가와 IDS 생산성의 증가를 확인하였다.

본 연구에서는 재조합 단백질 IDS의 생산성을 향상시키기 위한 방법으로 혈청이 첨가되거나 첨가되지 않은 배지에서 세포를 배양하는 동안 sodium butyrate를 첨가하였다. 두 배양 배지에서 모두 비록 세포 농도나 생존률이 감소되긴 했지만 생산성에서 상당히 높은 효과를 보였다.

본 실험에서는 골막조직 내에 존재하는 PDPCs를 분리하고, 기존 피부 이식을 위해 사용되는 $Hyalrograft^{\circledR}$ 3D에서의 chondogenesis가 가능함을 확인하였다. 수적인 확보가 제한되는 연골세포의 대체 세포로서 PDPCs의 가능성을 확인하였으며 동시에 $Hyalrograft^{\circledR}$ 3D에서의 연골화는 지지체의 양적 확보에 있어 기존의 $Hyalograft^{\circledR}$ C 보다 경제적일 수 있음을 의미한다.

This research was performed for developing of biological treatment process of odor gas such as MEK, $H_{2}S$, and toluene, which is generated from the food waste recycling process. To establish the operational conditions of odor gas removal by small-scale biofiltration equipment, it was continuously operated by using toluene as a treating odor object. When the odor treating microorganisms were adhered to fibril form biofilter, high removal efficiency over 93% was obtained by biofilm formation. At 400 ppm of inlet odor gas concentration and 10 sec of retention time, the removal efficiency was 76% and 93% in 1st stage reactor and 2nd stage reactor, respectively. However, the removal efficiency remained over 97% at the operational conditions above 15 sec of retention time.

3종의 생물흡착제에 대하여 5가지 중금속 이온 용액의 pH가 $4{\sim}8$일 때 크롬이온은 흡착이 감소하였고 구리이온은 흡착이 증가하였으며, 특정한 실험환경(pH4, $21^{\circ}C$, 120RPM교반)에서 교반시간이 60분일 때 흡착평형이 이루어졌으며 최대흡착율에 도달하였으며, Langmuir흡착 등온식이 Freundlich 흡착 등온식보다 더 높은 상관계수를 나타내며 단일-혼합 중금속 흡착 실험에서 카드뮴과 망간을 제외하고는 모두 80%이상, 납에서 95%의 흡착성능을 보였다.

The purpose of this study was to investigate the ultra sonic extraction method of phenolic compounds and antioxidant activity from Rumex crispus. Rumex crispus was fractionated with hexane, ethyl acetate, butanol and water. The amount of phenol compounds and antioxidant activity was presented higher content in ethyl acetate fraction then others.

This study was carried out to investigate the antimicrobial activity of Rumex crispus development of antibiotics derived from natural products. To confirm antimicrobial activity, paper disc method and growth inhibition in liquid culture were applied. Antimicrobial activity was observed in Saccharomyces cerevisiae, Vibrio vulificus, Escherichia coli, Bacillus cereus, Staphylococcus aureus, and Cαndida bombicola.

음식물 폐기물을 3단계 메탄발효 공정을 이용하여 처리하였을 때 배출되는 발효폐액은 거의 액상으로 고농도의 sCOD, T-N and $NH_{3}-N$를 함유하고 있다. 본 연구에서는 3단계 메탄 발효 후 최종 유출되는 고농도의 유기성 폐수를 물리${\cdot}$화학적 응집 및 침전을 이용하여 전처리 한 후 고효율 자외선/광촉매 시스템을 이용하여 후속 처리를 하고자 하였다. $FeCl_{3}$를 이용하여 전처리하였을 때 최적 pH 및 농도는 4와 2000 mg/L로 52.6%의 COD제거율을 보였다. 고효율 자외선/광촉매 시스템의 경우 24시간 동안 COD는 2890 mg/L에서 184 mg/L로 감소하였으며, T-N은 2496 mg/L에서 914 mg/L로 감소하였다.

Biofilm airlift reactor was continuously operated to investigate the competitions between the autotrophs and heterotrophs, ammonia oxidizers and nitrite oxidizers, and Nitrobacter and Nitrospira with real wastewater at a C/N ratio of 0.86. As the reactor achieved complete nitrification microbial distribution was analyzed by FISH/CLSM technique. The results showed that heterotroph was more abundant than nitrifying bacteria. Ammonia oxidizers (17%) and Nitrobacter (7%) prevailed nitrite oxidizers (9%) and Nitrospira (2%), respectively.

Hybrid시스템의 운전결과 황화수소의 removal efficiency는 황화수소의 inlet load가 $200g/m^{3}/h$에서 removal efficiency가 급격히 감소하여 약 60%의 수준을 유지하였다. 또한 톨루엔에 대한 removal efficiency는 거의 100%를 유지하다가 톨루엔의 inlet load가 $644g/m^{3}/h$로 증가할 때에는 약 60%로 감소하였다. 이와 같은 Hybrid시스템 운전결과로 부터 바이오필터만을 이용하여 황화수소와 톨루엔을 동시처리할 때보다 removal efficiency가 제고되었음이 관찰되었다.

Sugar polymers have been considered as biomaterial. Biomaterials are widely utilized for a medical applications in direct contact with living tissue Clearly, biomaterials must be carefully and microscopically fabricated for optimal acceptance within the living organism in both functional and structural senses. In this study, the enzymatic synthesis of sorbitan methacrylate from 1,4-sorbitan via the manipulation of an immobilized biocatalyst (Novozym 435) and acryl donors (methacrylic acid and vinyl methacrylate) was evaluated.

Bioconverted hydroxy fatty acid from ${\gamma}-linolenic$ acid showed antibacterial activity against Gram-positive bacteria such as Bacillus subtilis (ATCC 6633), Listeria monocytogenes (ATCC 19166), Staphylococcus aureus (ATCC 6538) and S. aureus (KCTC 1916) and one Gram-negative bacteria, Pseudomonas aeruginosa (KCTC 2004) with MIC ranging from 250 to $750\;{\mu}g/ml$ against five of eleven bacteria tested.

Bioconverted linolenic acid (bLNA) obtained from linolenic acid by Pseudomonas aeruginosa PR3, showed anti-fungal activity against plant pathogens such as B. cinerea, F. oxysporum, F. solani, P. capsici and C. capsici. The oil sample also showed anti-fungal activity with MIC values, ranging from >250 to >1,000 ${\mu}g/ml$. Varied concentrations of bLNA had a great potential effect on spore germination of different fungi.

L- arabinose residues are widely distributed in plant cell walls, where they are present in polymers such as arabinans, arabinoxylans, arabinogalactans and arabinogalactan proteins. L-arabinose suppress intestinal sucrase and decrease the adsorption of sugar in the small intestine, consequently, weight loss and fatness prevent. Now, xylose be used replacement sugar and arabinose be utilized fatness prevent of our time. Various Agricultural surplus like com fiber, contain $20\;{\sim}\;40%$ of hemicellulose. Corn fiber from Agricultural Renewable Biomass was chosen the best suitable material for arabinose production. In this work, we searched about for L-arabinose gene in compost, metagenome pool and indonesian soil. So, the B1029 TS2-8 of L-arabinase gene in compost was selected by YNB media(5% yeast nitrogen base, 5% arabinogalactan). After enzyme reaction with corn fiver, B1029 TS2-8 produced 2.15 g/L of L-arabonose.

Dihydroxy-acid dehydratase (DHAD, 2,3-dihydroxy-acid hydrolyase, EC 4.2.1.9) is one of the key enzymes involved in the biosynthetic pathway of the branched chain amino acid isoleucine and valine. Although the enzyme have been purified and characterized in various mesophiles including bacteria and eukarya, the biochemical properties of DHAD has bee not yet reported from hyperthermophilic archaea. In this study, we cloned, expressed, and purified a DHAD homologue from the thermoacidophilic archaeon Sulfolobus solfataricus P2, which grows optimally at $80\;^{\circ}C$ and pH 3, in E. coli. Characterization of the recombinant S. solfataricus DHAD (rSso_DHAD) revealed that it is the dimeric protein with a subunit molecular weight of 64,000 Da in native structure. rDHAD showed the highest activity toward 2,3-dihydroxyisovaleric acid among 17 aldonic acid substrates Interestingly, this enzyme also displayed 50 % activities toward some pentonic acids and hexonic acids when compared with the activity of this enzyme to the natural substrate. Moreover, rSso_DHAD indicated relatively higher activity toward D-gluconate than any other hexonic acids tested in substrates. $K_m$ and $V_{max}$ values of rSso_DHAD were calculated as $0.54\;{\pm}\;0.04\;mM$ toward 2,3dihydroxyisovalerate and $2.42\;{\pm}\;0.19\;mM$ toward D-gluconate, and as $21.6\;{\pm}\;0.4\;U/mg$ toward 2,3-dihydroxyisovalerate and $13.8\;{\pm}\;0.4\;U/mg$ toward D-gluconate, respectively. In the study for biochemical properties, the enzyme shows maximal activity between $70^{\circ}C$ and $80^{\circ}C$, and the pH range of pH 7.5 to 8.5. The half life time at $80^{\circ}C$ was 30 min. A divalent metal ion, $Mn^{2+}$, was only powerful activators, whereas other metal ions made the enzyme activity reduced. $Hg^{2+}$, organic mercury, and EDTA also strongly inhibited enzyme activities. Particularly, the rSso_DHAD activity was very stable under aerobic condition although the counterparts reported from mesophiles had been deactivated by oxygen.

The polysaccharides were extracted from fruiting body, mycelia, and cell-free broth of Agaricus blazei Murill. The crude polysaccharides were obtained by the ethanol addtion. They were further purified using ion-exchange chromatography and gel chromatography. Ion-exchange chromatography using DEAE-cellulose column separated neutral and acidic polysaccharides. Neutral polysaccharides were then purified with gel filtration chromatography. For single peak obtained from gel filtration chromatography was molecular weight was measured with Sepharose CL-6B. The same procedure with acidic polysaccharides were performed to get the purified polysaccharides.

한국산 개비자나무로부터 alkaloid계 항암 활성 물질인 Homoharringtonine의 분리 및 정제 공정을 개발하였다. 추출 용매로 메탄올을사용하여 biomass와 mthanol을 1:8의 비율로 분씩 3회 추출할 경우 개비자나무로부터 대부분(>99%)의 Homoharringtonine이 추출됨을 알 수 있었다. 흡착 공정에서는 활성 백토(active clay)를 사용하여 추출물에 포함되어 있는 식물유래 타르, 왁스 성분을 효과적으로 제거하였다. 크로마토그래피 공정을 통해서 52% 이상의 Homoharringtonine을 정제하였다.

분별침전 공정에서 순도 및 수율에 미치는 온도의 영향을 확인하였다. 분별침전을 위해 일정 온도에서 보관할 경우와 단계적으로 보관 온도를 하강하는 경우에 대한 분별침전 효율을 확인하였다. 일정온도$(0^{\circ}C)$에서 보관할 경우 높은 수율$({\sim}84%)$을 얻을 수 있는 반면 단계적으로 보관 온도를 하강할 경우 상대적으로 높은 순도$({\sim}79%)$를 얻을 수 있었다. 또한 단계적으로, 큰 폭으로 보관 온도를 하강 시킬 경우 더 높은 순도를 얻을 수 있었다. 저장온도$(-20^{\circ}C{\sim}12^{\circ}C)$를 일정하게 유지할 경우 paclitaxel 순도 면에서 $0^{\circ}C$가 가장 효과적이었으며 온도변화에 따른 수율은 거의 영향이 없었다. 따라서 최종 정제를 위한 전 처리 공정으로 분별침전 공정이 간단하고 매우 효과적임을 알 수 있었다.

This study was a method that used a hydrolysis for increasing the efficacy of alcohol decrease from Hovenia dulcis extract. The best pH was 2.0 to obtain a maximum activity at fixed reaction temperature and time. At pH 2.0, reaction temperature $80^{\circ}C$ and reaction time 4 hr gave the highest activity which was 124.2% of control. This is very simple and efficient method to increase the efficacy of alcohol decrease from Hovenia dulcis extract. The mechanism that increases the efficiency of alcohol decrease be examined through hydrolysis.

Phytoestrogens are non-steroidal compounds found in a variety of plants, which exert estrogenic effects in animals. In this study, the useful compounds of pomegranate as preliminarily research for the developing of natural estrogen supplement were determined. The estrogenic activity of phytoestrogens in the pomegranate was estimated by using the Yeast Estrogen Receptor and E-screen assay. Estrogenic activity of all pomegranate extracts in the Yest Estrogen Receptor assay were not significant difference at all concentration. Whereas peel extracts of Iranian and domestic red pomegranate are significantly enhanced in the E-screen assay. When various pomegranate extracts enzyme and acid hydrolyzed, three aglycones of phytoestrogen, kaempferol, quercetin and catechin were detected. Peel extract of domestic red pomegranste contained more than kaempferol(87.0 mg%), quercetin(172.8 mg%)) and catechin(956.8 mg%) than other extracts. These differences in concentrations of key phytoestrogens among various extracts seemed to be responsible for their differences in estrogenic activities. Among these three compound, kaempferol showed the highest MCF-7 cell enhancing efface.

The polyene antibiotics are a family of most promising antifungal polyketide compounds, typically produced by actinomycetes species. Using the polyene CYP-specific PCR screening with served actinomycetes genomic DNAs, Pseudonocardia autotrophica strain was identified to contain a unique polyene-specific CYP gene. The genomic DNA library screening using the polyene-specific CYP gene probe revealed the positive cosmid clone containing an approximately 34.5 kb DNA fragment revealed a total of seven complete and two incomplete open reading frame (ORFs), which are highly homologous but unique to previously-known polyene biosynthetic genes. These results suggest that the polyene-specific screening approach should be an efficient way of isolating potectially-valuable cryptic polyene biosynthetic gene cluster from various rare actinomycetes.

Lipomyces starkeyi produces a novel glucanhydrolase containing endo-dextranase and ${\alpha}-amylase$ activities. A cDNA from L. starkeyi encoding a dextranase was isolated and characterized. The 2,052 kb cDNA fragment (lsd1) carrying dextranase gene showed one open reading frame (ORF) composed of 1,824 bp flanked by a 41 bp 5'-UTR and a 184 bp 3'-UTR including a poly(A) tail of 27 bp. The ORF encodes for a 608 amino acid with a predicted molecular mass of 67.6 kDa. There was 77% deduced amino acid sequence identity between the LSD1 dextranase and the dextranase from Penicillium minioluteum. The primary structure of the dextranase from L. starkeyi has distant similarity with enzymes belonging to glycosyl hydrolase family 49. The lsd1 protein was expressed in the Saccharmyces cerevisiae under control of GAL1 promoter and active dextranase was produced.

대장균을 이용하여 형광단백질을 형질전환하고 배양하면서 그 특성을 관찰하고 형광세기 및 형광 단백질의 크기를 조사하였다.

TMC (Tautomycetin) is a liner polyketide immunosuppressive antifungal compound produced by Streptomyces spp. Inhibition of T cell proliferation with TMC was observed highly efficient at 100-fold lower than those needed to achieve maximal inhibition with cyclosporin A. To elucidate the biosynthetic pathway of TMC, a genomic DNA library was constructed using a E. coil-Streptomyces shuttle cosmid vector, pOJ446. The DNA libraries were screened by colony blot hybridization using several polyketide ${\beta}-ketosynthase$ (KS) probes amplified from TMC-producing Streptomyces genomic DNA using polymerase chain reaction (PCR), of which the degenerate primers were designed based on the highly conserved sequences present in KS domains of various type I polyketide synthase genes in Streptomyces species. This library construction and screening approach led to the isolation of several positive cosmid clones representing type I polyketide biosynthetic gene clusters. In addition, a Streptomyces regulatory gene called afsR2 (a global regulatory gene stimulating antibiotic production in both S. coelicolor and S. lividans) was successfully integrated into the TMC-producing Streptomyces chromosome via E. coil-Streptomyces heterologous conjugation mehtod. The more detailed results of production improvement and genetic characterization of TMC-producing Streptomyces spp. will be discussed.

Surface Plasmon Resonance(SPR) sensor system can be applicable for detecting of many biospecific interactions. In this study, the feasibility of the experimental $SPREETA^{TM}$ evaluation kit to analyze human IgG, Pseudomonas aeruginosa, was investigated. The sensor prepared for detecting of anti-human IgG has response on $0.1{\mu}{\ell}$ of the anti-human IgG solution. SPREETA was able to detect P. aeruginosa solution in the range above $10^8\;CFU/mL$ with the chitosan/ alginate multilayers.

In this study optical sensing membrane was developed for the queantification of dissolved carbon dioxide in micro-bioreactor using an immobilized 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS). For the immobilization of HPTS sol-gel was synthesied by using 3-glycidoxypropyl-dimethoxymethylsiline and tetraethyl orthosilicate.

This study was concentrated to investigate the dependency of recovery yield on the extraction conditions. From fruiting body, the extracted amount of polysaccharide was studied with various solvent. In order to maximize the yield, the optimum extraction conditions were elucidated with respect to solvent, temperature and time. In addition, the dialysis was applied to obtain the higher purity of polysaccharide with varying the exposure time.

6종 한약재의 70% 에탄올 추출물의 Helicobacter pylori의 위내 서식을 도와주는 것으로 알려진 urease의 활성억제능은 오매(Mume Fructus)의 에탄올(70%) 추출물에서 가장 높았으며, 소목 및 연교의 추출물도 높았다. 특히, 오매의 경우 한약재 추출물을 넣지 않은 대조군과 비교하여 90% 이상의 urease 활성억제 효과를 나타내었다.

조협은 trosinase 저해 활성 실험 결과 1.9%(w/v) 이상의 농도에서 70%이상의 저해를 나타내고, UV 흡수능에서는 220-230nm 에서 최대 흡수 파장이 나타나고 250-550nm 에서 고르게 흡수되는 경향을 보인다. 조협은 자외선 차단효과와 활성산소종 형성 억제 및 멜라닌 생성을 저해함으로써 피부미백 효과에 기여한다. 따라서 조협을 이용한 liposome을 제조하여 미백화장품에의 응용 가능성을 높여 줄 수 있다.

The purpose of this study was to determine the effect of rat intestinal ${\alpha}-glucosidase$ inhibitor ; methanol(80%), ethanol(80%) and $dH_2O$ extract of Schizandra chinensis in Korea(KS : Schizandra chinensis in Korea) and China(CS : Schizandra chinensis in China). When the final concentration was 1 $mg/m{\ell}$ for each sample(KS and CS), MeOH extract of KS($IC_{50}$ 1.62 mg/ml) showed 46.8%, EtOH extract of KS($IC_{50}$ 1.48 mg/ml) showed 47.4%, $dH_2O$ extract of KS($IC_{50}$ 1.72 mg/ml) showed 46.3% and MeOH extract of CS($IC_{50}$ 8.35 mg/ml) showed 13.3%, EtOH extract of CS($IC_{50}$ 8.05 mg/ml) showed 16%, $dH_2O$ extract of CS($IC_{50}$ 8.37 mg/ml) showed 11.54% of inhibiter for p-nitrophenyl ${\alpha}$, D-glucopyranoside ${\alpha}-glcosidase$ activity, respectively, And the contents of total phenol, flavonoid of Schizandra chinensis were measured. When the final concentration was 1 $mg/m{\ell}$ for each sample(KS and CS), total phenol and flavonoid in KS were higher than CS, respectively.

tyrosinase 저해 활성 실험 결과 1.2%(w/v) 이상의 농도에서 70% 이상의 저해를 나타내 색소침착에 대한 기능성 화장품 원료로 사용이 가능할 것으로 보이며, ACE 저해 활성 실험에서 1.5% 농도에서 60% 가량의 저해활성을 보여 혈관 변형 치료 및 예방에 효과가 있을 것으로 기대된다. UV 흡수능에서는 UV-B, UV-C의 영역에서 흡수 경향을 나타내며 자외선 차단제의 원료로 사용이 가능할 것으로 기대되며, 또한 bead 형성능을 가지고 있어 약물전달체로서 이용 가능성이 기대된다.

분무법을 이용한 alginate microsphere를 제조한 결과 구형을 형성하는 것을 관찰하였고, 이러한 alginate microsphere에서 alginate 농도가 증가할수록 크기는 작아지고, chitosan/alginate microsphere에서 chitosan의 농도가 증가할수록 그 크기가 증가하는 것을 확인하였다. OVA의 방출정도는 alginate microsphere에서 alginate 농도가 증가할수록 잘 이루어지지 않았고, HCl buffer에서보다 PBS buffer에서 방출이 잘되는 것을 확인하였다. Chitosan/alginate microspheres에서는 chitosan의 농도가 증가할수록 방출이 잘되지 않았고, 이는 alginate microsphere에서와 마찬가지로 PBS buffer에서 방출이 잘 이루어지는 것을 확인하였다.

본 연구에서는 유화법을 이용한 alginate 미세입자 제조에 있어서 적절한 alginate 농도와 가교제인 calcium chloride의 농도를 조사하였다. 유화법을 통해 calcium-alginate microsphere는 구형으로 제조되었으며 입자의 크기는 alginate와 가교제의 농도가 증가함에 따라 감소하는 경향을 보였다. 약물방출 경향은 alginate의 농도가 2%(w/v)일 때와 calcium chloride의 농도가 10%(w/v)일 때 가장 좋은 결과를 얻을 수 있었다.

The aim of this study was to prepare a hydrogels composed of alginate blended with a carboxymethyl scleroglucan (CMSC) and evaluate the feasibility of the hydrogels as a drug delivery system for a protein. The main advantage of the alginate/CMSC hydrogels is to improve a restricted drug release from alginate hydrogels. The CMSC was chemically synthesized with chloroacetic acid and confirmed using a FT-IR. The alginate/CMSC hydrogels were prepared at distinct compositions by crosslinking with calcium ions. The swelling ratios of these hydrogels increased significantly with increasing the content of CMSC. At pH 7.4, the swelling ratios of the hydrogels increased remarkably as compared to those at pH 1.2. In ovalbumin (OVA) release test, the amount of OVA released from the hydrogels showed higher as compared to those released at pH 1.2. In addition, the release of OVA was improved with increasing the content of CMSC. Thus, the alginate/CMSC hydrogels may be used as a potential system for oral delivery of protein drugs.