We have shown that it is possible to form a fibrilar network of fibronectin on a polyelectrolyte polymer film whose dimensions are similar to those reported on the extra cellular matrix. The fibronectin network was observed to form only when the charge density of the polymer was in excess of the natural charge density of the cell wall. Furthermore, the self-organized fibronectin layer was much thicker than the polymer film, indicating that long ranged interaction may playa key role in the assembly process. It is therefore important to understand the structure of the polymer layer/protein interface. Here we report on a neutron reflectivity study where we explore the structure of the polyelectrolyte layer, in this case sulfonated polystyrene (PSSx,), with varying degree of sulfonation (x<30%), as a function of sulfur content and counter ion concentration. These results are then correlated with systemic study of the adsorption and the multilayer formation of fibronectin as a function of incubation time for various sulfonation levels of $PSSx.^1$

Although Dacron and ePTFE have most widely been used for artificial vascular grafts, these materials cannot be used for small-diameter grafts (l.D.<6mm) due to thrombotic occlusion. To overcome this limitation, a small-diameter vascular graft was developed with stem cell and tissue engineering method. Autologous bone marrow stem cells were cultured and seeded onto small-diameter (4mm) collagen tubular matrices. The matrices were anastomosed to carotid arteries in canine models. Prior to implantation, histological and electron microscopical examination revealed stem cell adhesion and growth on the matrices. Angiography indicated that the vascular grafts maintained patent for 8 weeks. Histological examination showed the regeneration of endothelium, media and adventitia in the grafts. This study may allow us to step forward to the development of tissue-engineered small-diameter vascular graft appropriate for clinical applications.

Cartilage defects are common and painful conditions that affect people of all ages. Although many techniques have developed, none of the current available treatment options is satisfactory. Recent advances in biology and materials science have pushed tissue engineering to the forefront of new cartilage repair techniques. The purpose of this study is to determine effective regeneration method for tissue-engineered cartilage. A serum free medium was developed for cartilage tissue engineering. Chondrocyte passage number was found to influence greatly on cartilage tissue formation in vivo. Injectable, biodegradable polymer matrix was developed for chondrocyte transplantation through injection. Transplantation of chondrocytes mixed with the injectable matrices resulted in the cartilage formation in nude mice's subcutaneous sites and rabbit knees. This study may lead to the development of tissue-engineered cartilage appropriate for clinical applications.

Mammalian cell based biosensor kits are expected to be in assessment of samples toxicity more sensitive and accurate. A recombinant fluorescent Chinese Hamster Ovary (CHO) cell line was known to be responsive to the various toxicants Specially. KFC- AlO cell line. which contain the c-fos SRE::GFP plasmid (pKFG). was found to be able to detect toxicants sensitively. A biosensor kit was developed by using an immobilized KFC-A10 cell line. Immobilized recombinant fluorescent cells within agarose, known as a representative hydrogel matrix, have been maintained in the matrix viably and have shown constant fluorescent levels for long time. Immobilized cells have shown the ability to detect the chemical toxicity in the keep of fluorescent level as the metabolism is inhibited under toxic conditions.

현재의 반월판 연골손상의 치료법인 부분 봉합술이나 동종이식은 부작용이나 공여자의 부족 등의 많은 문제점이 있다. 따라서 이러한 치료법을 대체할 생체조직공학기술을 이용한 새로운 반월판 연골 재생술이 필요하다. 이 연구에서 생분해성 합성고분자인 PGA와 반월판 연골세포를 이용하여 재생한 조직공학적 반월판 연골은 실제의 반월판 연골조직과 유사함을 확인하였다. 조직공학을 이용한 반월판 연골재생은 반월판 연골손상에 이상적인 치료법이 될 수 있을 것이다.

We have previously shown that the addition of silkworm hemolymph to a culture medium increases the longevity of insect and mammalian cells by inhibiting apoptosis. This indicates that the component which inhibits apoptosis is contained in the silkworm hemolymph, The apoptosis-inhibiting component was isolated from silkwonn hemolymph and characterized in our previous study. A database search using the N-terminal amino acid sequence of this component as a template resulted in a 95% homology with a low molecular weight lipoprotein, the so called ’30K protein' of unknown function. In this study, the 30K protein gene was expressed in mammalian and insect cells to confirm the apoptosis-inhibiting effect. The overexpression of 30K protein in mammalian cell inhibited the staurosporin-induced apoptosis by the prevention of the activation of caspase 3. Using an Autographa californicanuclear polyhedrosis virus (AcNPV) system, the 30K protein was overexpressed also in insect cells. The expression of the 30K protein increased the longevity of baculovirus-infected insect cells by inhibiting apoptosis. These results suggest that the 30K protein is a novel anti-apoptotic protein.

SPAD공법은 탈질을 위한 전자공여체로 입자상의 황과 소량의 메탄올을 이용하는 생물학적 공법이다. 이번 연구에서 SPAD공법은 특정 폐수에의 적용가능성을 평가하기 위해 고농도의 칼슘이온을 함유한 울산의 S특수강 폐수(200-300$NO_3\;^-$ -N/L)에 적용되었다. 운전기간은 2001년 11월부터 2002년 3월 초까지였으며 반응조 내부의 온도는 약 $20^{\circ}C$ 부근으로 유지되었고, 그러한 낮은 온도조건하에서도 탈질 효율은 90%이상으로 유지되었으며 유출수 농도는 약 20mg $NO_3\;^-$ -N/L정도로 유지되었다. 따라서 SPAD공법은 고농도의 칼슘이온을 함유한 폐수에 대해 적용이 가능한 것으로 사료되어진다.

Thermus caldophilus로부터 TDP-glucose 4,6-degydratase (TDPDH) 유전자를 분리하고 염기서열을 결정하고, 결정된 염기서열에 해당하는 유전자를 발현벡터에 삽입하고, 발현벡터를 대장균에 형질전환시켜 배양함으로써 TDP-glucose 4,6-dehydratase를 대량 생산하였다. 본 연구의 TDP-glucose 4,6-dehydratase는 주반응으로 TDP-4-keto-6-glucose를 합성하고, 여러 가지 당을 디옥시당으로 합성하는 효과가 있다.

Arachidonic acid is a polyunsaturated fatty acid(PUFA) containing twenty carbon atoms with four double bonds. The family of w-6 PUFA, including arachidonic acid as well as r-linoleic acid, was served as intermediates in the formation of several key prostaglandin and leukotrienes. Several fungal strains of the genus Mortierella accumulate high amounts of arachidonic acid. In this study experiments were carried out to optimize the culture conditions for the mass production of fungus Mortierella alpina DSA -12 and lipid production with high proportion of polyunsaturated fatty acids, especially arachidonic acid. The batch culture was carried out in 500 L fermenter containing 50 g/L glucose, 18 g/L corn-steep powder and 100 mg/L MnS04 under $25^{\circ}C$, aeration rate of 0.5 vvm and agitation speed of 200 rpm without pH control. As a result, we could be obtained 22 g/L of cell mass with high contents of lipid 12.1 g/L) and arachidonic acid (5.1 g/L) The intermittent fed-batch culture was performed in the medium containing 20 g/L glucose and 10 g/L corn-steep powder. The final glucose concentration was 170 g/L and pH was maintained at 5.5 ${\sim}$ 6.0 by adding 14% ammonia solution. It was shown relatively high cell concentration (70.5 g/L) with high contents of lipid (45.8 g/L) and arachidonic acid 08.3 g/L). Therefore, when compared to batch cultures, the high concentration of arachidonic acid could be obtained by fed-batch culture using M. alpina DSA -12. These results imply that the fed-batch culture of M. alpina DSA -12 was feasible in industrial purpose and could be employed in the commercial production of arachidonic acid.

A gradient LC/MS system was constructed and applied for separation of biological samples. For example, a rapid and simple analytical method without pretreatment based on gradient ${\mu}LC/MS$ with a disposable microcolumn has been developed to determine B group vitamins in urine. Urine samples were directly injected to the disposable home-made microcolumn. The microcolumn can be emptied after being used for a series of urine samples, and repacked with fresh stationary phase. An overdose of vitamin pills were swallowed by healthy volunteers and the urine samples were taken 1,2,3,5, and 8 hours after swallowing. Vitamins immediately showed up in urine, hit the maximum, and disappeared swiftly. This technique is expected to have some application for clinical purposes.

본 연구에서는 균주의 배양과 동시에 생산물을 회수할 수 있는 동시추출공정을 건조질량의 $15{\sim}75$ %의 탄화수소를 생산한다고 알려진 B. braunii 배양에 적용하고자 한다. 일반적인 tow-phase 동시 추출공정의 적용시 B. braunii의 경우 생산된 탄화수소가 균주 외벽의 matrix에 강하게 부착되어 있기 때문에, two phase 추출공정 적용시 bubble solumn내에서 단지 폭기에 의한 교반만으로는 충분한 탄화수소의 회수율을 얻을 수가 없었다. 본 연구에서는 배양액과 유기용매층의 접촉기회를 증대시킨 two-stage 동시추출 공정을 개발하여 기존의 two-stage 동시추출 공정보다 2배 이상 높은 57 %의 탄화수소 회수율을 얻을 수가 있었고, 이를 회분배양후 후속분리공정으로 이용할 경우 6시간 추출후 62 %의 회수율을 얻을 수가있었다.

Rhodococcus rhodochrous IGTSS (ATCC 5396S) can break organo sulfur compounds such as dibenzothiophene. Since the environment for biodesulfurization process is invariably hydrophobic, parameters in hydrophobic systems should be examined. For the model oil, hexadecane-containing 5.43mM dibenzothiophene, the volumetric desulfurization rate was decreased with the oil-to-aqueous phase ratio up to 50%. The rate declined sharply after 48h because the cell activity, which is refreshed by medium exchange, was lost. To supply the exhausted nutrients, medium exchange was performed. At 30% oil phase, most of DBT was removed by medium exchange on 48h, and the rate was 2.03mg $DBT_{removed}/L_{dispersion}-hr.$ At 50% oil phase, medium exchange on 60h was performed and the rate was 1.79mg $DBT_{removed}/L_{dispersion}-hr.$ The 300mL flask system was scaled up to a 5-L bioreactor system. On 60 h, a medium exchange was performed and the rate was 5.28mg $DBT_{removed}/L_{dispersion}-hr.$ and all of DBT was removed. It means that we can use the biodesulfurization process even 10 the high oil-to-water phase by some appropriate methods such as controlled feeding of key nutrients and the dilution or removal of some toxic metabolites by continuous reactor.

Monascus pigment-dipeptide derivatives were synthesized by Monascus in 48-well plates. Monascus pigment-dipeptides derivatives were identified as antibacterial agents. The antibacterial activities against 20strains were tested with ELISA Reader and 96-well plate.

Monascus pigment was produced by Monascus species. During Monascus fermentation, citrinin, the mycotoxin was produced with pigments. Citrinin can become a problem for use of monascus pigment as a food colorant. We found adding of S. cerevisiae filtrate during Monascus cultivation could enhance production of red pigment whereas it could reduce citrinin level. When we added the filtrate at 24 hand 48 h, respectively, pigment production increased about 400% and citrinin concentration decreased to 30%. In a glucose medium, there was no special effect by addition of filtrate. On the other hand, the effect was striking in a sucrose medium.

Acetobacter xylinum KJ1 efficiently producing bacterial cellulose(BC) in shaking culture was isolated from a rotten grape. The strain was used to investigate optimum operating conditions for increasing BC production and factorial design model was employed for the optimization. The results of experiments were statistically analyzed by SAS program. Reciprocal effects of each factors(carbon source concentration, shaking speeds(rpm), oxygen pressure, and CSL concentration) and culture condition of BC production were examined by getting regression equation of the dependent variable. Comparisons between experimental results and predicted results about BC concentration were done in total 24 experiments by combination of each factors using SAS program, and the correlation coefficients of BC concentration and BC yield were 0.91 and 0.81, respectively. The agitated cultures were performed in various operation conditions of factors which affected considerably to BC production in jar fermentor. The results showed that BC concentration was 11.67g/ L in 80 hours cultivation under the condition of carbon source concentration shaking speeds(rpm) : oxygen pressure: CSL concentration = 4% : 460rpm : 0.28 : 6%. On the other hand BC yield was 0.42g/g in 80 hours cultivation under the condition of carbon source concentration shaking speeds(rpm) : oxygen pressure: CSL concentration = 4% : 564rpm : 0.21 : 2%. The BC production could be enhanced up to more than 65.3% by factorial design. The result of a verifying experiment under the optimal conditions determined by the factorial design to the BC production showed that the model was appropriate by obtaining BC concentration of 11.02g/L in the optimum condition

In this work we describe the on-line monitoring technique for the analysis of fumaric acid in biotechnological processes. Fumarase and malate dehydrogenase(MDH) were immobilized on epoxy carrier and integrated into a FIA system. The effects of carrier buffer flow rate, pH, reaction temperature on the immobilized fumarase/MDH were investigated for the development of a fumarate-FIA system. Furthermore the effects of substrates, salts and metabolites dissolved in the sample on the activity of the immobilized enzyme were investigated.

Two types of alginate gel beads capable of floating in the gastric cavity were prepared. The first, alginate gell bead containing olive oil(Al-Oil), is a hydrogel bead and its buoyancy is attributable to olive oil held in the alginate gel matrix. The model drug, metronidazole(MZ), contained in Al-Oil was released gradually into artificial gastric fluid. The profiles of MZ release from Al-Oil shown initial burst and after 90 min they were about 100%. The second, alginate gel bead containing curdlan microsphere(Al-C), is a gel bead with curdlan-MZ microsphere in the matrix. To sustained release rate of drug, alginate bead were prepared curdlan microsphere containing MZ. Results demonstrated that sustained delivery of MZ over 2h can be easily achieved while the bead remained float. The release properties of prepared alginate beads are applicable not only for sustained release of drugs but also for targeting the gastric mucosa.

막 생물반응기를 이용하여 일정한 TMP 와 일정한 플턱스 여과방식에 따른 실험을 수행한 결과, 일정한 TMP 에서보다 일정 한 투과 플럭스에서 여과하는 것이 여과의 초기 단계동안 과도한 오염을 피할 수 있으므로 바람직하다는 것을 확인하였다. 투과 플럭스를 고정시킨 실험에서 속도에 대한 임계 플럭스 증가 간격은 선형적이었으며, supra-critical zone 과 sub-critical zone 의 두 지대로 구분되었다. 막 생물반응기는 화학적 세정 없이 장 기간 운전되어야하므로 TMP를 고정시키는 깃보다 플럭스를 고정시켜 운전하는 것이 효과적이여, 최적운선조건은 sub-critical zone 경계층 위, 임계플렉스 바로 아래상태임을 알 수 있었다.

생균제로 새로이 개발된 효모인 Pichia amomala를 대량 생산하기 위한 배양 실험으로 가격이 저렴하고 균주 생육에 적절한 질소원으로의 CSL 실험을 실시한 결과 Yeast extract 에 비해 보다 우수한 생육을 보였고 , CSL 을 이용한 3ton fermenter 에서의 batch 실험결과 최대 OD 58까지 배양되었고 , batch 배양시 harvest 시점은 가장 많은 viale cell 을 얻을 수 있는 시점인 배양 후 20 시간으로 결정되었다 . 또한 원심분리후 동결건조시 안정제에 따fms 동결건조시의 생존에 대한 실험을 실시한 결과 skim milk 2%, scrose 2% 에서 가장많은 생균수를 얻을 수 있었다.

TLC(Thin Layer Chromatography) 는 제약산업이나 생화학 연구 그리고 여러 산업 현장에서 널리 이용되는 화학 분석 방법이다 . 본 연구에서는 적은 비용으로도 많은 양의 시료를 신속하게 분리할 수 있는 TLC를 이용하여 유기산인 젖산 (Lactic acid) 을 분리는 전개방법을 개발하였다 . 전개용매는 2 가지 용매 (1) nitroethane nitromethane : ethanol: water: I-propanol = 1 : 2 : 3 : 4 : 5 (v/v/v/v/v), (2) diisopropyl ether. formic acid. water = 90 : 7 : 3 (v/v/v) 를 사용하였다 . 발색시약은 A : bromocresol purple reagent I, B . bromocresol purple reagent II, C : bromocresol green-bromophenol blue-potassium permanganate reagent 를 사용하여 분석하는 방법을 개발하였다 $^6)$. 젖산의 분리는 silica gel TLC plate를 이용하는 경우 , 용매 (1) 에 발색시약 B를 사용했을 때 , 분리 확인이 가장 좋았다.

Effect of carbon source and culture conditions involved in the concentration of dissolved oxygen on cell growth and the production of pullulan by A. pullulans HP2001 were investigated. Among those carbon sources, glucose was found to be the best carbon source for the production of pullulan by A. pullulans HP2001. Maximal production of pullulan by A. pullulans HP2001 was 26.6 g/ f when concentrations of glucose and yeast extract were 8% (w/v) and 0.25% (w/v), respectively. It was found that aeration rate, agitation speed and inner pressure of a bioreactor, which were some of physiological factors involved in the dissolved oxygen in the medium may affect cell growth and the production of pullulan by A. pullulans HP2001.

The process for the production of mannitol with fructose (5% to 25%) using Leuconostoc mesenteroides NRRL B-1149 was investigated. Optimization study for mannitol production was carried out in 8 liter batch or fed-batch cultures at $28^{\circ}C$, pH 5.0, without aeration. When 5% fructose was used in a batch culture fermentation, the yield of mannitol was 78% of theoretical. As the concentration of fructose was increased to 10% in a batch culture, the yield was reduced to 59.6% of theoretical. Using a fed-batch fermentation with 10% fructose, the yield was increased to 81.9%. When 15% fructose was used for a fed batch fermentation 5% fructose was initially added and the last 10% fructose was supplied continuously. The final yield of mannitol was 83.71% of theoretical. When 20% fructose was used, the yield was more higher, 89.48%.

Pseudomonns putida BCNU171 had the tolerant ability to several other organic solvents headed by toluene and xylene. Several mutants were made by mating of BCNUl71, pJFF350 to clarify the structure tolerance gene related. From this mutants the 7 of mutants related with toluene sensitive mutants were selected. pBCNUT-2, pBCNUT-4, pBCNUT-9 was transformed, and from this separated plasmid DNA sequences the gene having high homology was searched. In the case of toluene sensitive mutant it was todX gene (pBCNU4) related with cell membrane, ttgE gene (pBCNU2, pBCNU9) and ttgF gene (pBCNU2, pBCNU9).

Lovastatin is a cholesterol-lowering agent, which plays a role of an inhibitor of 3-hydroxy-3- methylglutaryl coenzyme A reductase (HMG-CoA). When thiamine was supplemented in 3L batch fermentation, the production of lovastatin was improved. At the same time, the levels of pyruvic acid and NAD(P)H were estimated in the course of the fermentation of A. terreus. For the high level production of lovastatin, semi fed-batch fermentation was performed. And the thiamine level was maintained to a concentration of 20 mg/L and glucose was supplied. The final dry cell weight was lowered by 30 % and final lovastatin concentration was increased by 33 %. Final lovastatin concentration of 3.3 g/L was achieved in 8 days.

A glucan and a fructan producing enzymes from Leuconostoc mesenteroides NRRL B-1149 were prepared and concentrated from the cu1ture of 1.5% sucrose using polysulfone ultrafiltration hollow fiber in the presence of 0.1% (w/v) Tween 80, 1 mM $CaCl_2$, and 0.02% $NaN_3$. The molecular masses of the enzymes were estimated to be about 213.6 kDa and 180 kDa, respectively, based on the PAS staining for the glucosyltransferase and Mukasa method for fructosyltransferase. Polymers produced by the enzymes showed different solubility; an insoluble glucan and a soluble fructan. The linkages of polymers were determined by methylation using Hakomori reagent and following acid hydrolysis. The glucan was composed of ${\alpha}$-1,6 and 1,3 linkages and the fructan showed similar linkage data of levan.

Effect of aeration rate and agitation speed on cell growth and the production of heteropolysaccharide-7 (PS-7) by Beijerinckia indica was investigated. Aeration rate and agitation speed in a 7L bioreactor ranged from 0.5 to 1.5 vvm and from 300 to 500 rpm, respectively. Higher agitation speed with an aeration rate of 0.5 vvm in the bioreactor resulted in maintenance of higher concentration of dissolved oxygen in the medium, which enhanced the production of PS-7. In this study with a 7L bioreactor, maximal production of PS-7 was 11.0 g/L and its conversion rate from 2% (w/v) glucose was 0.55 when the aeration rate and agitation speed were 1.0 vvm and 500 rpm, respectively. Proper aeration rate and agitation speed might enhance the production of PS-7 as well as reduce the time to reach maximal production.

Compositions of four kinds of products inhibit aging and protect many kinds of disease through eliminating. These compositions inhibit growth of harmful bacteria. such as Bacteroides fragilis. Clostridium perfringens. Staphylococcus aureus, Listeria monocytogenes. Vibrio parahaemolyticus etc. Bacteroides fragilis was controlled by mixing SCUTELLARIA BAICALENSIS GEORGE+CRATAEGI FRUCTUS+PAEONIA JAPONICA+SCHIZANDRAE FRUCTUS. SCUTELLARIA BAICALENSIS GEORGE+CRATAEGI FRUCTUS+PAEONIA JAPONICA+CORNI FRUCTUS. etc. and Clostridium perfringens was SCUTELLARIA BAICALENSIS GEORGE+PHELLODENDRI CORTEX+EPIMEDII HERBA+ASTRAGALI RADIX. SCUTELLARIA BAICALENSIS GEORGE+EPIMEDII HERBA +ASTRAGALI RADIX+GLYCYRRHIZAE RADIX.ste. Listeria monocytogenes was NELUMBO NUCLFERA GAERTNER+SCUTELLARIA BAICALENSIS GEORGE+COPTIDIS RHIZOMA+PAEONIA JAPONICA, NELUMBO NUCLFERA GAERTNER+SCUTELLARIA BAICALENSIS GEORGE+COPTIDIS RHIZOMAtSOPHORAE FLOS. etc. and Vibrio parahaemolyticus was ACANTHODANACIS CORTEX +ASTRAGALI RADIX+PHELLODENDRI CORTEX+COPTIDIS RHIZOMA ACANTHODANACIS CORTEX +ASTRAGALI RADIX + PHELLODENDRI CORTEX+PAEONIA JAPONICA. We expected it as a functional food economical and easy type of taking.

gellan 생산의 최적 pH 조건을 알아보기 위해 삼각플라스크와 회분식 발효를 이용 하여 실험하였다. 삼각플라스크 내에서 초기 pH 7.0 은 1.66g/ ${\ell}$ 의 gellan 을 생산하였고 , 회분식 발효에서는 pH 5.5-8.5 의 범위에서 pH를 조절하면서 실험한 결과, pH 3.0 에서 균체량 O.48g/ ${\ell}$f) 및 gellan 생산량O.97g/ ${\ell}$) 이 최대값을 나타내었다.

효모 Pichia ciferrii 어 l 의해 생산되는 TAPS 는 쉽게 탈아세틸화 시킴으로 ceramide로 전환이 가능한 물질로 palmitoyl-CoA 와 아미노산인 L-serine으로부터 합성된다. 따라서 TAPS 생산에 영향을 줄 것으로 예측되는 serine과 serine 합성에 관련된 amino acid 들의 TAPS 생산에 대한 영향을 조사한 결과 cysteine 은 TAPS 생성에 방해를 주었으며 serine 은 TAPS 의 전구체임에도 불구하고 TAPS 농도의 향상을 기대할 수 없었다. 그러나 serine 의 전구체가 되는 glycine 의 첨가는 약 25% 의 TAPS 농도의 증가를 보여주었으며 특별히 glutamate 의 첨가는 20% 의 cell mass 의 증가와 37.7% 의 가장 많은 TAPS 의 증가를 보여주었다.

Baker's yeast was cultured with $Na_2SeO_3$. Selenium compounds in yeast were extracted and analyzed by size exclusion chromatography. Selenium was broadly distributed in the fraction of protein. For the inhibition test of MMP-l induction, selenium containing compounds was fractioned by ultrafiltration

The optimal condition for the production of DXAMase, containing the both characteristics of dextranase and amylase, was studied based on different levels of pH, temperature, and aeration rate. Response surface methodology was applied to find the optimatic condition showing the relationship between the fermentation response(dextranase and amylase activity of DXAMase) and the fermentation variables(pH, temperature, and agitation rate). In case of dextranase activity, the condition of pH 4.06, $28.08^{\circ}C$, and 235.14 rpm showed the highest activity, 2.26 U/ml, and for amylase activity, the condition of pH 4.01, $27.96^{\circ}C$, and 212.01 rpm showed the highest activity, 3.52 U/ml. For the production of DXAMase, dextranase and amylase, the optimum condition was pH 4.06, $28.08^{\circ}C$, and 234.80 rpm.

식물병원균에 대한 Helicosporium 의 항균작용을 확인하기 위하여 식물병원균인 Rhizoctonia solani, Rhizoctonia solani AG2-2, Fusarium oxysporium, Phytophthora dreschler, Alternaria 속을 선정하였고, 이에 대한 항균작용을 검토한 결과, Rhizoctonia solani, Rhizoctonia solani AG2-2 에는 강한 항균활성을 나타내었으며, Alternaria에는 미약한 항균활성을 가졌으며 Fusarium oxysporium, Phytophthora dreschler에 대해서는 항균활성을 하지 않는 것으로 확인되었다. 이 항균물질의 구조를 $^1H-NMR$ 스펙트럼을 통해서 분석한 결과 이 물질은 콜레스테롤 에 해당하는 구조를 가진 것으로 추정된다.

본 연구에서는 자체 제작한 2L 규모의 bubble-column photobioreactor에서 H. pluvialis의 배양을 시도하였고 astaxanthin의 축적량 증가를 유도하기 위하여 배양시작 20일 후에 light stress를 주었다. 이 방법을 통하여 대조구에 비하여 68%의 건조균체중량 증가와 215%의 astaxanthin 축적량 증가를 유도할 수가 있었고, bubble-column photobioreactor를 이용한 유도배양이 H. pluvialis의 배양에 적합함을 알 수가 있었다.

Colistin produced from Penibacillius polymyxa was widely used as an antibiotic active against gram-negative bacteria and as feed additive. This research studied on increment of colistin productivity by mutation of P. polymyxa. As a result, several mutants were obtained from the strain by UV radiation and NTG treatment. They produced approximately 8.5${\sim}$9.0 g/L of colistin in flask and jar culture. Colistin productivity of the mutant, named Penibacillius polymyxa CBY, showed 100 times than that of wild type. When Penibacillius polymyxa CBY fermented in the optimal medium, it produced up to 18 g/L of colistin in jar fermentation.

Itaconic acid has been produced during the cutivation of Aspergillus terreus DSMZ 5770 by using several starchs as carbon sources. The starchs were pretreated by partial hydrolysis with some acids at various pH conditions. The highest yield for the production of itaconic acid has been found when rice starch was pretreated by sulfonic acid at pH 2.5 and utilized for the cultivation. Using the results from shaker fermentation A. terreus has been cultivated in 2.5 L bioreactor for the production of itaconic acid and its on-line monitoring.

공업적으로 중요성이 날로 더해가는 phospholipase C (PLC) 를 Bacillus cereus를 이용하여 생산하였다. 또한, plc::gfp fusion protein 을 생산하는 재조합 E. coli 를 제조하고 배양하였으며 특히 형광센서를 이용하여 PLC 의 생산 특성을 모니터링하였다.

Leuconostoc mesenteroides B-512FMCM, 742CB3, 1299C의 dextransucrase들의 glycosyl기 전이 특성을 수용체 반응과 transglycosylation반응을 통해 확인하였다. 수용체 반응의 경우 10% sucrose에 수용체로 4% maltose를 첨가하여 반응시켰고 transglycosylation반응은 다른 크기, 다른 농도 그리고 다른 종류의 가지 결합의 dextran 을 합성하는 효소들을 이용하여 수행하였다. 각각의 효소들은 maltose를 이용한 수용체 반응에서 유사한 종류의 수용체 산물들을 합성한 것에 비해 세 dextransucrase들 (512FMCM, 742CB3, 1299C) 을 일정 비율로 혼합하여 maltose를 이용한 수용체 반응 결과 512FMCM 효소의 활성 비율을 줄이고 742CB3, 1299C 효소의 활성 비율을 증가시켰을 경우에는 ${\alpha}-1{\rightarrow}$3 의 가지결합이 많은 dextran 을 합성하였다. 또한, 세 가지 다른 구조의 dextran(T40, 742CB, B1299)에 100mM maltose을 수용체로 첨가해 각각의 dextransucrase(512FMCM, 742CB3, 1299C)와 transgly cosylation을 수행한 결과 1299C 효소가 세 종류의 dextran(T40, 742CB, B1299) 에 모두 가지 결합이 많은 dextran을 합성함을 확인하였다. 또한 ${\alpha}-1{\rightarrow}$6 결합으로 주로 이루어진 2%, 5% dextran(T10, T40, T7O, T500, T2000)에 dextransucrase(512FMCM, 742CB3, 1299C)를 반응시켜 기존의 dextran 보다 가지 결합이 더 많이 형성된 transglycosylation 산물을 합성하였다. 이때 maltose를 첨가했을 경우 이 수용 체에 의해 많은 ${\alpha}-1{\rightarrow}$6 가지 결합의 dextran 을 합성함을 확인하였다.

We have synthesized branched oligosaccharides (BOS) by the mixed-culture fermentation (MBOS), fructosyltransferase (FBOS) or glucosyltransferase (GBOS) with high concentration of sucrose (3M). MBOS was further modified as iron and sulfate-oligosaccharides. The modified MBOS were stable at high temperatures (up to $140^{\circ}C$) and low pHs (2 to 4). Most highly branched and modified oligosaccharide (0.34%, w/v) effectively inhibited fructose release from sucrose by Streptococcus mutans 6715 mutansucrase. FBOS, GBOS, iron-MBOS inhibited the mutansucrase activities from Streptococcus sobrinus about 46.8%, 49.2% and 43.1%, respectively. Most highly branched and modified oligo- saccharides (0.5%, w/v) effectively inhibited the fonnation of insoluble glucan and adherence of S. mutans or S. sobrinus cell in the presence of sucrose. Modified oligosaccharides affected the growth and acid production of oral pathogens. Cytotoxicity test showed that highly branched and modified oligosaccharides was non-toxic.

In this study, biological production of fumaric acid by Rhizopus oryzae KCTC 6946 using rotary biofilm contactor was investigated. In study of neutralizing agent on fumaric acid production, $Na_2CO_3$ was more effective than NaOH. After 24 hr of incubation with a rotating speed of 10 rpm at $35^{\circ}C$, biofilm was grown on and around the surface of the disks. The yield and volumetric productivity of rotary biofilm contactor were 33.8% and 0.595 g/L${\cdot}$h, respectively, with the optimum effective disk surface area of 1,583 $cm^2/L$.

본 연구에서는 형질전환된 P. pastoris를 이용한 재조합 HBsAg 생산에서 유가식 배양시 공급배치 탄소원으로 sorbitol이 단백질 발현에 미치는 영향을 glycerol과 비교하여 실험하였다. 유가식 배양 공급배지 탄소원으로 50% sorbitol을 이용했을 때 50% glycerol을 이용하는 경우보다 세포 증식 측면에서는 methanal 유도 후 균체 농도가 낮은 경향을 보였으나 이것은 glycerol 이 sorbitol 보다 에너지원으로써 높은 affinity를 가지기 때문인 것으로 판단된다. 하지만 단위 건조 균체량당 단백질 발현량은 50% sorbitol을 공급 한 경우 50% glycerol을 공급한 경우 보다 12% 향상되는 결과를 보였다. 따라서 유가식 배양용 공급 탄소원으로 sorbitol을 이용했을 때 glycerol을 이용하는 것보다 AOX promoter에 의한 단백질 발현에 보다 긍적적인 효과가 있는 것을 확인 할 수 있었다.

For the antimicrobial activity test, Akebia quinata DECAISNE stem were extract water and fractionate it with four solvents which had a different polarity, antimicribial activity were increased in oder of petroleum ether fraction = diethyl ether fraction ${\leq}$ n-buthanol fraction < ethyl acetate fraction < aqueous fraction. Water extracts of stems and fruits were showed inhibitory two species bacteria](Bacillus subtills KCTC 1021, Bacillus cereus KCTC 1012) from developing.

Dissolved oxygen level of cell culture media has a critical effect on cellular metabolism, which governs specific productivity of recombinant proteins and mammalian cell growth However, in the cores of cell aggregates or cell-immobilized beads, oxygen level frequently goes below a critical level. Mammalian cells have a number of genes induced in the lower level of oxygen, and the genes contain a common cis-acting element (-RCGTG-), hypoxia response element (HRE). By binding of hypoxia inducible factor-l (HIF-I) to the HRE, promoters of hypoxia inducible genes are activated, which is a survival mechanism. In this work, to develop a CHO cell capable of producing recombinant proteins in immobilization and high density cell culture efficiently, mammalian expression vectors containing human tissue-type plasminogen activator (t-PA) gene controlled by HRE were constructed and stably transfected into the CHO cells. In $Ba^{2+}$ -alginate immobilization culture, CHO/pCl/dhfr/2HRE-t-PA cells produced 2 folds higher recombinant t-PA activity than CHO/pCl/dhfrlt-PA cells without $CoCl_2$ treatment. Furthermore, in repeated fed batch culture, productivity of t-PA in immobilized CHO/pCI/dhfr/2HRE-t-PA cells was 121 ng/ml/day, total production of 0.968 mg/day at 11 days culture while CHO/pCIIdhfrlt-PA cells was 22.8 ng/ml/day. All these results indicate that HRE is very useful for the enhancement of protein productivity in mammalian cell cultures.

CHO (Chinese hamster ovary) cells were transfected with plasmids containing both cis-acting HRE (hypoxia response element) and CMV-promoter that controls tissue-type plasminogen activator (t-PA). CHO cells with HRE produced 16.2 fold higher t-PA concentration than CHO cells without HRE. It was noted that hypoxia strongly induced CHO cell apoptosis. which resulted in decrease of cell viability and protein production. In this study. by introducing Bcl-2, anti-apoptotic gene, we tried to recover cell viability and increase the protein production. When batch culture of both control cells without transfection of Bcl-2 and cells transfected with Bcl-2 were performed in the absence of CoCl ι hypoxia mimic condition. the cells with Bcl-2 were effected specific cell growth rates, maximum cell density. Immunoblotting assay showed Bcl-2 was recombinant with HRE dependent t- P A expression cassette, and their expression level was depended on hypoxia. By introducing Bcl-2, both cell viability and maximum cell density could be increased.

Lactate and ammonia are two major toxic waste products formed during mammalian cell culture. Accumulation of the side products have negative effects of on cell growth and specific production rate. In this study, K-562 cells were used as the host cell of a recombinant protein. Effects of carbon sources were invetigated focused on the cell culture span, the accumulation of lactate and ammonia in culture of recombinant K-562 cells.

Oxygen is a key substrate in animal cell metabolism and its consumption is thus a parameter of great interest for monitoring and control in animal cell culture bioreactor. The use of a gas-permeable membrane offered the possibility to provide the required quantity of oxygen into the culture. while avoiding problems of foaming or shear damage generally linked to sparging. For determining the optimum DO control strategy of this gas-permeable membrane aeration bioreactor, the oxygen transfer rate coefficient was measured with varying $N_2$ ratio in inlet air. The results showed that an increasing mass flow rate of nitrogen reduced the $K_La$ value. and 5% nitrogen in air did not result in any oxygen limitation.

Newcastle disease virus (NDV) vaccines were produced from Vero cells by using lively attenuated virus strain. The MOI of 0.1.' serum concentration of 2%. initial pH of 8.0. and infection time of 3 days were found to be optimum conditions for vaccine production. The treatment of polycation enhanced the virus production. When ascorbic acid was added as an antioxidant, NDV production was also enhanced. Utilization of $CaCl_2$ showed an inhibitory effect on the propagation of NDV. It was also found the ammonium ion concentration higher than 4mM inhibited virus production. Thus ammonium ion removal system was tried for the efficient production of NDV vaccine.

Specific pathogen free (SPF) eggs have been used to produce live vaccines. however, their application causes many problems such as cost, space and waste disposal. The substitution of mammalian cells for SPF eggs offers a desirable system of vaccine production. In this study, mammalian cells were tested for the infection of Newcastle disease virus (NDV). As a result, DF-I and MDBK cells showed high virus productivity compared to the other mammalian cells. For the highest productivity of NDV, the optimal multiplicity of infection (M.O.I.) in DF-I or MDBK cells was determined to be 0.2 or 0.5 M.O.I., respectively.

This study was conducted to optimize phytohormones combination and concentration on resveratrol production of Vitis vinifera callus cultures. TDZ was so effective for callus proliferation and resveratrol production and can be expected as a stimulus to the cells which keep the ability of producing resveratrol. We optimized the hormone combination of NAA 0.5 mg/L and TDZ 2 mg/L to stimulate resveratrol production very effectively. And callus under MS medium with the optimized phytohormones combination was treated by fungal elicitor, Botrytis.

The corneal tissue consists of three layers : epithelium, stroma, and endothelium. Central cornea is a highly differentiated tissue whereas the limbus contains the epithelial stem cell. In the present study. we report the engineering of the three-dimensional reconstructed cornea derived from rabbit limbal epithelial and stromal cells. The differentiation degree of corneal stem cells were assessed in serum concentration and inoculation density of stromal cells. Optimal condition differentiation of corneal stem cells is achieved when 5% FBS was supplemented to culture medium and $1-2{\times}10^5$ cells/ml inoculation density of stromal cells.

Hyperosmotic pressure increased specific antibody productivity ($q_{Ab}$) of recombinant CHO cells (SH2-0.32) while it depressed cell growth. Thus, the use of hyperosmolar medium did not increase the maximum antibody concentration substantially. To overcome this drawback, the feasibility of biphasic culture strategy was investigated. In the biphasic culture, cells were first cultivated in the standard medium with physiological osmolality(294 mOsm/kg) for cell growth. When cells reached the late exponential phase of growth, the spent standard medium was replaced with the fresh hyperosmolar medium (522 mOsm/kg) for antibody production. The ($q_{Ab}$) in growth phase with the standard medium was 2.1 ${\mu}g/10^6cell/day$ while the ($q_{Ab}$) in antibody production phase with the hyperosmolar medium (522 mOsm/kg) was 11.1 ${\mu}g/10^6cell/day$. Northern blot analysis showed a positive relationship between the relative contenet of Ig mRNA and ($q_{Ab}$), indicating that transcriptional regulation was involved in the response of rCHO cells to hyperosmotic pressure. Due to the enhanced ($q_{Ab}$) and increased cell concentration in biphasic culture, the maximum antibody concentration obtained in biphasic culture with 522 mOsm/kg medium exchange was 161% higher than that obtained in batch culture with the standard medium. Taken together, simple biphasic culture strategy based on hyperosmotic culture for improved foreign protein production from rCHO cells is effective in improving antibody production of rCHO cells.

감미제로 많이 사용되고 있는 약용식물 중 대추와 감초 황기로부터 기능성 친연 감미료를 얻기 위해 에탄올과 물을 추출용매로 사용하여 당류가 포함된 시료를 얻었다. 이들 시료에 대한 감미효과를 구체적으로 알아보기 위해 당 분석 및 난충치성 실험을 실시한 결과 황기와 감초에서 산생성력과 dental plaque의 수치가 낮게 나타나 기능성 천연 감미료를 얻을 수 있는 약용식물로 판단된다.

참당귀 (Angelica gigas Nakai) 현탁세포의 고농도 배양을 위하여 perfusion 배양을 한 결과 세포의 생장이 증진됨을 확인하였다 . 배양 5 일째부터 sucrose를 4배 농축한 배지로 2 mL 씩 교환해준 결과, 세포의 생장이 계속 유지되었고 최대 세포농도도 23.7 g/L 로 대조구에 비하여 1.7 배 증가하였다. 초기 당농도를 50 g/L 로 하여 배지 교환을 한 결과, 최대 세포농도도 23.8 g/L 로 30 g/L 당농도 대조구에 비하여 1.6 배 증가하였고, 배지교환을 하지 않은 50 g/L 대조구에 비하여 1.1 배 증가하였다 . 따라서 초기 당농도를 높이고 연속적으로 배지교환을 해준다면, 고농도의 세포를 얻을 수 있으며 결과적으로 참당귀 세포가 생산하는 다당류의 생산량을 증진시킬 수 있을 것이다.

접종 초기 root segmentation 은 뿌리의 생장 속도를 감소시키고 그 생산성을 저하시키기도 하지만 F/D ratio를 감소시켜 뿌리가 자랄 수 있는 배지 내 공간을 제공하기 때문에 고농도 배양에 유리할 뿐 아니라, 생산성이 높은 secondary root의 비율을 증가시키기 때문에 decursinol angelate 의 생산량을 중대시킨다. 참당귀 뿌리배양을 위한 plant growth regulator의 최적 조건으로 secondary root의 생장성 측면이나 decursinol angelate 의 생산성 측면에서 4 mg/L 이상의 IBA 첨가가 요구됨을 확인하였다.

GS system을 이용하여 재조합 EPO를 생산하는 새로운 CHO-K1 세포주를 확립하였으며, 이 세포주를 이용하여 Zn 이온과 Mn 이온이 EPO의 생산에 미치는 영향에 관해서 연구하였다. 형질 전환에 있어서 DNA 3 ${\mu}g$ 올 사용하고, 세포군 선별을 위한 MSX의 농도는 100 11M 을 사용한 경우에만 세포군이 발견되었다. 200 ${\mu}M$의 MSX를 처리한 경우에서는 세포군이 생성되지 않았고, 이는 고농도의 MSX에 의해 세포 생장이 저해되었기 때문이다. 형질 전환된 CHO-K1 세포 배양에 Zn 이온을 첨가한 결과 세포의 생장은 크게 저해되지 않은 반연에, EPO의 생산은 대조구에 비해 40% 이상 증가함을 알 수 있었다. 이러한 결과는 Mn 이온을 처리한 실험에서도 관찰할 수 있었다.

형질 전환된 식물세포배양에서 비이온성 계면활성제인 Pluronic F-68 의 첨가가 미치는 영향을 연구하였다. Pluronic F-68 첨가시 세포생장에 미치는 영향은 크지 않았으며 5 g/L 첨가시 배지내 hGM-CSF의 양은 대조구에 비해 2배 더 증가하였다. 첨가 시기 최적화 실험을 통하여 배양 초기부터 첨가하는 것이 유리할 것으로 판단된다. 또한 Pluronic F-68 의 첨가가 세포크기를 감소시킴을 알 수 있었다. 이는 Pluronic F-68 이 세포막과 상호 작용하여 세포막 투과성을 증진시킴으로써 생산된 hGM-CSF의 배지 내로의 분비를 촉진시킴을 간접적으로 보여주는 결과이다.

10 가지 식물추출물에서 항산화활성을 측정하여 가장 효과가 높았던 오미자를 선택하여 순차적으로 층분리 후, 각 유기용매층에 따라 분리된 추출물의 항산화 활성을 측정하였다. 그 결과 Ethyl acetate층 추출물 (DPPH 51.6%. SOD 66.2%)과 Butanol층추추물 (DPPH 60.7%, SOD 67.4%) 에서 가장 높은 항산화활성을 나타내었으며 총 polyphenol 함량 또한 Ethyl acetate층 (30000 mg/kg) 과 Butanol층 (30000 mg/kg)에서 높게 나타났으며, 특이한 점은 항산화 활성이 낮았던 water층 (39400 mg/kg) 에서 총polyphenol함량이 높았다.

본 연구에서는 hGM-CSF를 생산해 배지 내로 분바하도록 유전자 조작된 Nicotiana tabacum 세포를 수성이상계 내에서 배양하여 in situ recovery를 시도하였다. 특히 식물세포 자체의 생상에 큰 영향을 주지 않으면서도 일반 배지에서 회수한 단백질에 비해 많은 양을 생산할 수 있는 수성이상계 시스템을 선정하였다. 6% (w/w) PEG 20,000과 10% (w/w) dextran 2.000,000 을 이용하는 경우 최대 세포 농도가 18.6 g/L 로, 일반배지를 사용하는 경우 (15.7 g/L)에 비해 저해가 없음을 확인하였다. 또한 목적 단백질인 hGM-CSF의 생산에 있어서도 위의 시스템의 경우 dextran-rich phase인 아랫 상으로 분배됨으로 인해 회수가 쉬울 것으로 확인되었으며 그 생산량 또한 일반배지에서의 생산량 (1.5 ng/mL)에 비해 크게 다르지 않음을 확인할 수 있었다. 따라서 hGM-CSF의 회수를 위한 in situ recovery 시스템에 있어서 수성이상계의 이용 가능성을 확인하였다.

Sucrose 의 경우 저농도에서는 후반부로 갈수록 세포생장 속도는 현저히 감소하는데 비해 hGM-CSF 생산은 후반부에 급격히 증가함을 확인하였다. 고농도 sucrose를 사용하는 경우에는 lag phase가 길어지는 동안에 hGM-CSF의 생산이 증가하였다. 따라서 배양 초기에는 고농도 sucrose가, 배양 후반에는 저농도 sucrose로 존재하는 경우에 hGM-CSF를 많이 얻을 수 있었다. Nitrogen source의 농도는 60.52 mM과 121.04 mM일 때가 세포의 생장이나 hGM-CSF의 생산을 증가시켰으며, phosphate의 경우에는 4.96 mM 일 때가 대조구인 2.48 mM 일 때보다 hGM-CSF의 생산을 3 배 증가시켰다.

The root of Panax ginseng C. A. Meyer is the one of traditional medicines used for many therapeutic purpose in the orient for many years. Polysaccharides isolated from ginseng root were known for mitogenic, antitumor and hypoglycemic activities. We studied the production of ginseng polysaccharides from ginseng hairy roots and compared with natural ginseng root.

Activated charcoal(AC) is generally used in plant tissue culture. Its addition to plant tissue culture was known to many advantages and disadvantages. We investigate that sucrose hydrolysis, which by autoclaving with or without activated charcoal on different intitial pH and sucrose concentration and that the effect of activated charcoal on plant tissue culture.

This study was performed by the airlift bioreactor using the nitrifier consortium entrapped in polyvinyl alcohol(PVA) for removing low concentration total ammonia nitrogen(TAN). At the aeration rate of 1.5 vvm, TAN removal rate and removal efficiency was 316.6${\pm}$7.2 $g/m^3$ day and 92.8${\pm}$2.2%. Removal rate was continuously increased with decreasing from 0.5hr to 0.05hr of hydraulic residence time(HRT), whereas removal efficiency was decreased with decreasing HRT.

This study estimated the effect of influent TAN concentration. temperature and pH in the airlift bioreactor(aeration rate; 1.5 vvm, HRT 0.35hr) using immobilized nitrifiers by PVA. At the effect of influent TAN concentration, removal rate was increased with increasing it and removal efficiency maintained 93${\pm}$2%. The optimum temperature for nitrification was $30^{\circ}C$ and at this point. removal efficiency was 95.5${\pm}$1.5%. It was effective to nitrify at $10^{\circ}C$ of low temperature. In the pH range from 7 to 9 in the bioreactor. removal rate and removal efficiency was 310${\pm}$10 $g/m^3$ day and 94${\pm}$3%.

햄은 부패에 의한 폐기율이 높은 식품으로서 본 연구에서 개발한 식물유래 천연방부제들 즉 향나무, 대나무, 소목, 쑥 등의 알콜추출물의 첨가에 의한 실용화 가능성이 높았다.

부산광역시 수영 하수처리장의 소화조에서 농축조로 보내지는 혐기성 슬러지를 탈기된 증류수와 1:1로 희석하여 11.900 mg/L로 만든 후 혐기성 고정 생물막 반응기에 15일간 생물막을 부착시킨 후, 부유 슬러시를 제거하고 각 반응기에 각각 8.00 mgTOC/L, 9.76 mgTOC/L, 및 18.97mgTOC/L의 기질 농도를 유입하여 HRT 0.496일로 각 반응기에 연속적으로 주입하여 실험하였다. 기질 전달 현상과 관련하여 각 반응기에 대한 실험 결과는, 저농도로 기질이 유입된 반응기 l과 2에서는 생물막 두께 및 기질 제거율, 기질 소비 속도 상수($k_v$), 유효 확산 계수($D_{eff}$) 가 비슷하였으나, 고농도로 기질이 유입된 반응기 3에는 자농도로 기질이 유입된 반응기 1과 2 보다 높은 값을 나타내었다. 이는 본 실험에 사용된 혐기성 미생물이 고농도의 기질을 유입하였을 때, 더욱 원활하게 성장함에 따라 높은 기질 소비를 나타내었다.

본 연구는 제약폐수 처리에 있어서 폐수처리장내의 미생물 활성을 최대로 하여 운전효율을 높이는 것을 목적으로 한다. 생물 반응기내의 미생물의 활성과 최적 용존산소 농도에는 어떤 상관관계가 있는가를 규명하기위해 연속식 실험을 진행한 결과, 0${\sim}$1.0 ppm 정도의 낮은 DO농도에서는 COD의 감소율도 적고, 1.5${\sim}$3.0 ppm 정도일 때에는 COD의 감소율이 다른 DO농도일 때 보다 상대적으로 높았다. 그러나 3.0 ppm 이상의 DO농도에서는 오히려 COD 감소율이 작아진다.

1. TPA 제거능이 우수한 미생물을 개발하였으며 동정 결과 Corynebacterium sp. 로 판단되며 Corynebacterium sp. YT-14로 명명하였다. 2. TPA 제거를 위한 Corynebacterium sp. YT-14의 최적 배양온도는 $35^{\circ}C$ 였고, 최적 초기 pH는 pH 10이었다. 3. Stirred loop bioreator에 Corynebacterium sp. YT-14를 배양한 후 감량가공폐수와 종합염색폐수를 회분식으로 처리한 결과 감량가공폐수에서 처리 7일 후 TPA 제거효율이 85.4% 였으며, 종합염색폐수에서는 처리 3일 후 TPA가 검출되지 않았다.

본 연구에서는 오존과 생물활성탄을 연계시킨 공정을 통하여 용존 유기물질의 제거 경향을 살펴보았다. 오존 처리를 거치면서 원수중의 난분해성 용존 유기물질과 같이 생물학적 분해 속도가 느린 화합물의 상당량이 저분자 형태로 전환되었음을 파악할 수 있었으며, 오존 처리후의 생분해성의 향상에 의해 흡착의 부담이 경감되어 활성탄의 수명이 연장되는 것으로 조사되었다. 생물활성탄 반응조 내에서 여층 깊이에 따른 DOC 제거 경향을 알아본 결과, 전체 제거량의 약 50%가 column 상단에서 제거되었으며, 따라서 짧은 EBCT 에서도 용존 유기탄소의 제거는 용이한 것을 사료된다. 또한 유기 오염에 대한 지표로서 국내외에서 일반적으로 고도정수처리의 처리 대상물질로 선정되어 있는 암모니아성 질소의 제거는 75.9%로 상당히 높은 제거율을 나타내었다.

인산가용화균 Penicillium sp. PS-113의 포자를 고체분말비료로 제제화 시 장기보존함과 동시에 균의 생활력 유지를 위하여 고체배양 후 건조공정(60, 80, 100, $120^{\circ}C$을 달리하여 수분 함량을 10, 15, 20%로 각각 조절하여 시험한 결과, $4^{\circ}C$에서 2개월 저장시 $80^{\circ}C$에서 건조하여 수분함량을 15%로 조절한 경우가 가장 효과적이었으며 초기에는 건조하지 않은 경우에 비해 일시적으로 생존율이 다소 떨어지나 저장기간이 길어질수록 생존율이 60배 이상 증가하였다.

염색폐수 중에 포함된 PVA를 생물학적으로 제거하기 위하여 PVA 분해용 미생물 50종을 분리하였다. 분리된 균주의 PVA 분해효율을 살펴보기 위하여 단일균주만을 이용한 실험과 단일균주들의 조합을 사용하여 실험을 진행하였는데, 단일균주를 사용했을 때는 최대 60%, 조합을 사용했을 때는 최대 96%의 분해율을 얻을 수 있었으며 3일 이내에 80% 이상을 분해하였는데, 이러한 결과는 지금까지 발표된 연구에서 보고된 PVA 분해기간 중 가장 빠른 결과이다.

군포 공단 주변 슬러지를 미생물 접종원으로 무기염배지에 10g/ ${\ell}$ 의 sucrose를 첨가하여 수소 생산 균주 Ye 13-6을 분리하였다. 분리 균주 Ye 13-6은 호기성조건과 혐기성 조건에서 모두 생장하는 통성 혐기성 균주였다. 유기성 폐기물 내에 다량 함유되어있는 glucose와 sucrose의 농도변화가 수소 생산 속도 및 수소 생성효율에 미치는 영향에 대하여 알아보았다. Glucose를 1${\sim}$12g/ ${\ell}$ 의 범위로 첨가할 경우 lag phase 없이 생장하였으며, 첨가량이 증가할수록 수소 비생산속도가 증가하여, 12g/ ${\ell}$ 에서 $60mmol-H_2\;{\cdot}\;mg-DCW^{-1}\;{\cdot}\;h^{-1}$의 최대값을 나타내었으며, 수소 생산 수율은 2.6 ${\sim}$3.1 $mol-H_2\;{\cdot}\;mol-glucose^{-1}$의 범위였다. Sucrose를 $1{\sim}12g/\;{\ell}$ 의 범위에서 첨가할 경우 약 10시간의 lag phase 후 원활한 생장을 보였다. 비생산속도는 및 수소 생산수율은 sucrose 첨가량이 증가할수록 증가하여 각각 $163mmol-H_2\;{\cdot}\;mg-DCW^{-1}\;{\cdot}\;h^{-1}$ 및 $4.5\;mol-H_2\;{\cdot}\;mol-glucose^{-1}$의 최대값을 보였다.

기계적 성질이 우수한 PVC와 생분해성이 우수한 고분자로 알려져 있는 폴리카프로락톤 (PCL)과 블렌드하여 새로운 소재의 생분해성 필름을 제조하여 생분해성 효과에 대해 조사하였으며, 그 결과 PCL/PVC 필름의 표면은 8주 후에 다수의 작은 구멍이 형성되었으며, 이러한 결과는 PCL의 함량이 9%로 낮아도 생분해성을 지닌다는 것을 의미한다.

Response surface methodology (RSM) was successfully applied to optimize for the production of Ganoderma lucidum in batch fermentations using the whey (40,000 mg latose/L) as substrate. This study was performed according to the central composite design (CCD) with respect to pH and temperature, where the designed intervals were 3.3$22.9^{\circ}C$$37.1^{\circ}C$, respectively. A second-order factorial design of the experiments was used to build empirical models providing a quantitative interpretation of the relationships between the two variables. The optimum conditions to maximize the production of G. lucidum were pH 4.2 and $28.3^{\circ}C$. At optimum conditions, the mycelial dry weight (MDW) and residual soluble COD (SCOD) were simultaneously used to evaluate the biokinetic coefficients assocoated with substrate inhibition model by nonlinear least squares method with 95% confidence interval. The. maximum microbial growth rates (${\mu}m$), half saturation coefficient ($K_s$), and the inhibition substrate concentration ($K_{is}$) were determined to be 0.095 l/hr, 128,000 mg SCOD/L and 49,000 mg SCOD/L, respectively. And the microbial yield coefficient (Y), biomass decay rate coefficient ($K_d$), and the maintenance energy coefficient ($m_s$) were determined to be 0.37 mg MDW/mg SCOD, 0.001 1/hr, and 0.0015 1/hr, respectively.

식물의 토양전염병을 유발하는 P. aphanidermatum 및 R. solani에 대한 길항력, 생장속도 및 표면장력 감소를 고려한 RPI (relative performance indices) 기법을 이용하여 90 여개 후보균 중 #16 길항균을 선정하였다. 선정된 Bacillus sp. #16은 16S rRNA 서열을 분석한 후 GenBank에서 비교한 결과 B. subtilis DSM10과 99% 유사성을 보였다. 또한 pot test 결과 오이 모잘록병에 대해 우수한 방제가를 보였다.

BTEX가 함유된 groundwater에 에탄올이 첨가된 경우 토양 미생물에 의한 BTEX의 분해는 에탄올이 존재하지 않는 경우에 비해 크게 낮아짐을 알 수 있었다. 이는 토양내 미생물이 에탄올을 우선 이용하며, 따라서 토양내 산소와 mineral의 결핍을 야기하여 BTEX의 분해가 느려짐에 기인한다. 전자수용체로 Fe(III), nitrate, sulfate를 groundwater에 첨가한 경우, BTEX의 분해도는 크게 증가하였으며, sulfate의 효과가 가장 높았다.

Bioremediation has been showing promise as an alternative to conventional environmental cleanup technologies. The objective of this study is to maximize the degradability of jet fuel in the soil system. The cells isolated from petroleum contaminated site was used for the degradation of jet fuel. When this strain was cultured in the MSM(minimal salt media) containing jet fuel for ten days, the degradability of jet fuel was almost 100%. The concentration of jet fuel did not affect the degradability much and the increased inoculution of strain and addition of nitrogen source decreased the time for complete degradation of jet fuel in the liquid culture. Inoculation of this strain increased the jet fuel degradability in the soil column by 15% and the aeration(50ml/min) and the addition of nutrients($NaNO_3$, $KH_2PO_4$) enhanced the jet fuel degradability(about 90%).

Industrial wastewater with high ammonium concentration was treated in batch biological systems which was a modified Ludzack- Ettinger process. Up to 78% conversion of $NH_4\;^+-N$ to $NO_x\;^--N$ was achieved in batch culture condition. Under anoxic condition with methanol as the carbon source, the denitrifiers decreased $NO_x\;^--N$ concentration from 608 mg/L to 5.6 mg/L in 22 d. As well as anoxic denitrification of $NO_x\;^-$ to $N_2$, dissimilatory nitrate reduction to ammonium also occurred under the condition as respiratory denitrification.

This experimental shows the possibility of using as biofertilizer, which convert insoluble inorganic phosphate salts to plant-usable phosphate type by immobilized microorganism with calcium alginate. In the case of culture of P. agglomerans on constant medium pH, phosphate was produced 357 mg/L after 18hrs. And in the case of culture of immobilzed P. agglomerans bead, phosphate was produced maximum 295.6 mg/L after 120 hrs. Also as using rock phosphate as insoluble phosphate salts, phosphate was respectably produced 190.3 and 195.2 mg/L after 36 hrs at free cells and immobilized cells. In our experiments, the using soils contained 23.16 g-P/kg-soil total phosphate and 3.76 g-P/kg-soil soluble phosphate. The result of 1g immobilized bead seeding, soluble phosphate was produced maximum 6.14 g-P/kg-soil phosphate and this value was increased continuously.

Strain YJ는 혐기 발효에서 glucose를 이용하여 복합 유기산 및 수소를 생산하였다. 2% glucose를 이용할 때 총 38ml의 수소가 생산되었으며, pH 완충제인 인산염을 첨가시켰을 경우 1.21배의 수소가 증가되었다. 4% sucrose를 기질로 이용하였을 때 총 40.5ml의 수소를 생산하였으며, 5% fructose에서 38.1ml의 수소를 생산하였다. Glucose를 이용할 때 이 균주는 butyrate, propionate를 많이 축척하였고, acetate, formate등이 소량 검출되었다.

Acylated homoserine lactones (AHLs) are membrane-permeant signal molecules responsible for biofilm formation of gram-negative bacteria via a unique mechanism known as quorum sensing. A target specific bioassay employing the AHL-responsive Agrobacterium tumefaciens reporter strain has been developed to identify new AHL-like compounds from natural products, which could be developed into antifouling compounds. By varying the X-gal concentration, incubation time, solvent for sample preparation and the sample loading procedure, it was possible to detect low level AHLs up to $10^1nM$. The length of the acyl chain of the AHLs was found to affect the sensitivity of this bioassay.

A performance of pilot scale biofilter was investigated for the treatment of odorous waste gas. The system treated 508.7 $m^3h^{-1}$ of waste air with amine and ammonia concentrations ranging up to 0.06 $N-gm^{-3}$ Over the four month study, its elimination capacity was demonstrated up to 6.37 $N-gm^{-3}h^{-1}$ and its removal efficiency was 100%. Differ to lab scale biofilter, It is difficult to maintain the proper temperature of reactor because of low external temperature. Experiments were performed indicating various problem such as sudden system shut down, unstable pump, water addition malfunction and corrosion of reactor. However, pilot scale biofilter was stable when treatment of odorous waste gas.

Fermentative hydrogen production by Citrobacter sp. Y 19 was investigated in batch culture. Optimal hydrogen production activity was observed at pH 6 - 7 and temperature of $36^{\circ}C$, and hydrogen yield and maximal hydrogen production rate were 1.12 mmol/mmol glucose and 32.3 mmol/g cell${\cdot}$h, respectively. With glucose as a substrate, the bacterium produced ethanol, acetate, and carbon dioxide as major glucose fermentation by-products. Y19 could utilize various sugars such as galactose, fructose, lactose, sucrose, and starch for cell growth and hydrogen production.

A novel process for $H_2S$ gas treatment has been introduced, based on the combined action of a chemical absorption step and a biological step involving the biocatalytic activity of the bacterium Thiobacillus ferrooxidans. The aim of this study is the development of a process for $H_2S$ elimination from gas streams based on that chemical/biological method. The immobilized biomass reactor/chemical adsorption system is suitable for application of the removal of $H_2S$. A double stage reactor was used for the experimental work. The removal efficiencies of over 99% were observed in the range of inlet $H_2S$ concentration from 200 to 1,000ppm. The novel process showed the stable elimination efficiencies of over 95% under the retention time range from 20 to 40sec at the 1,000ppm of $H_2S$ inlet concentration.

본 실험실에서는 음식폐기물을 이용하여 단시간내에 메탄을 생산할 수 있는 Pilot 규모 (2.5 톤 )의 3단계 메탄 발효 공정을 개발하여 운전하고 있다. 3단계 메탄 발효공정 가운데 첫 번째 단계인 반혐기성 가수분해/산발효조에서 유기물 분해균주를 11개 분리하여 그 특성을 조사하였다. 분리된 균주는 gram 양성균이 8 개 균주, gram 음성균이 3개 균주이며, catalase에 대해서는 모두 양성반응을 보였고 모두 간균의 형태이었다. 분리된 균주들 가운데 amylase 와 protease의 활성은 분리균주 K5이 다른 균주들에 비해 매우 우수하였다.

단일균이나 활성슬러지를 이용하여 폐수를 처리하고자 할 때 폐수의 pH 가 단일균이나 활성 슬러지의 최적 pH 에 적합하지 아니한 경우에는 미생물을 고정화하여 희망하는 pH에서 최대의 분해활성을 나타내도록 최적 pH를 어느정도 변화시킬 수 있으며, 기존의 활성슬러지법 보다는 활성 슬러지를 고정화하여 처리하는 것이 높은 부하에서도 제거 효율이 높기 때문에 반응조의 콤펙트화가 가능할 것으로 사료된다.

일정한 유량에 대해서 시간이 경과할수록 제거율은 약간씩 증가함을 알 수 있었으며, 이것 은 시간이 지난수록 내부세공막힘 현상이 유발되어 오염물질 자체가 제거율을 높이는 역활을 한 것으로 생각된다. 전처리한 시료에 분맞활성탄을 주입하지 않은 경우의 COD거율은 매우 낮게 나타났으며, 분말활성탄을 주입한 경우는 유기물이 분말활성탄에 흡착하여 96%의 매우 높은 제거율을 보였다. 압력에 따른 투과플럭스 및 제거율에 미치는 영향은 세가지 시료 모두 압력이 증가할수록 투과플럭스가 증가하였다. 또한 일정한 압력에 대해서 시간이 경과할수록 투과플럭스는 현저히 감소하였으며, COD 제거율은 압력이 증가할수록 약간씩 감소되었다.

본 연구의 목적은 기존의 단일 균주보다 두 종의 서로 다른 균주를 복합시켰을 때 두 균 주간의 상호 보완적인 면을 이용하여 효소생산을 극대화하기 위함이며, 다음과 같은 결론을 얻었다. 실험에서 사용된 T. viride와 FB01는 최적 pH와 온도가 비슷하여 혼합배양이 가능 하였으며, 이들의 복합균주를 개발하는데 성공하였다. 두 균주간의 상호작용으로 인해 단일 균주일 때보다 CMCase, ${\beta}-glucosidase$ 및 avicelase의 활성이 우수했으며 또한 계대배양 및 pH 조절을 통해 ${\beta}-glucosidase$이 활성이 최고 3.2배까지 증가함을 알 수 있었다. 탄소원으로 섬유소 폐기물중 특히 볏짚을 이용했을 때 복합균주에 의한 효소생산이 효과적이었으므로 앞으로 이 조건에서 복합균주를 장기적으로 계대배양하면서 효소의 생산성 향상과 복합균주의 안정성을 관찰할 필요가 있다고 사료된다.

The Pseudomonas fluorescens KCTC 1767, a selected and identified as potential candidate for stereo-specific resolution of rac-ketoprofen ethyl ester, was systematically investigated in order to induce the high level expression and detailed characterization of the expressing enzyme esterase. We cloned the esterase gene from chromosomal DNA of Pseudomonas fluorescens KCTC 1767 by PCR with two synthetic primers that desinged for simple purification. The recombinant esterase from Pseudomonas fluorescens KCTC 1767 exibited a high conversion rate and enantioselectivity to the (S)-ketoprofen ethyl ester as expected. The enzyme was easily purified to homogeniety by using a metal chelating affinity chromatography as a protein with poly histidine taq, and thus obtained 0.6 mg of protein from a 100 mL culture broth in a single step. The purified enzyme was steadily stable at the pH range from 7.0 to 10. The activity was also retained to be about 70% after the preincubation at $40^{\circ}C$ but over $50^{\circ}C$ lost the activity completely. The molecular mass of the esterase was estimated to be about 43 kDa on SDS-PAGE, and an identical result was also shown in gel filteration chromatography. The specific activity was calculated 27 mM/mg-protein/min by using the rac-ketoprofen ethly ester as a substrate.

Trichoderma sp. FJ1의 cellulase 생산을 위한 배지조성의 검토에서, 질소원으로서 0.1% bacto peptone을 사용하였을 때 생산성이 향상되었으며, 순수 상업용기질인 avicel과 섬유소 폐기물인 볏짚의 혼합배양에서 높은 효소생산성이 얻어졌다. 경제적인 효소생산성을 위한 기질로 섬유소 폐기물의 농도와 혼합비를 검토한 결과, 1%농도에서 볏짚과 펄프를 50:50으로 사용했을시 CMCase, xylanase, ${\beta}-glucosidase$, avicelase는 24.3, 38.7, 1.5, 0.6 U/ml가 얻어졌다. 이러한 효소생산성은 타 보고서의 결과보다 동등이상의 우위를 보여주고 있으며 섬유소 폐기물의 생물학적 당화기술에 크게 기여하리라 사료된다.

Trichoderma sp. FJl의 섬유소 분해효소 생산조건을 최적화하기 위해 반응표면 분석법을 이용하였다. 반응표면 분석을 위한 실험 계획법은 중심합성법 계획법을 이용하였으며, 주요 배양인자로서 탄소원의 농도, 질소원의 농도, 혼합탄소원 비율, 그리고 배양시간에 대해 조사하였다. CMCase의 경우에서는 탄소원의 농도 3.5%, 질소원 농도 0.6%, 혼합탄소원인 avicel 및 CMC의 비율 52:48, 그리고 배양시간 5.4일에 33.5 U/ml로 최적화되었고, xylanase의 생산은 3.5%, 0.8%, 54:46의 배양조건에서 5.3일에 52.6 U/ml이었고, ${\beta}-glucosidase$의 생산은 5.0%, 1.0%, 83:17의 배양조건에서 7 일에 2.88 U/ml이었고, avicelase의 생산은 4.0%, 0.9%, 64:36의 배양조건에서 6.5일에 1.84 U/ml로 최적화되었다. 이중에서 ${\beta}-glucosidase$의 생산성은 기존 실험조건보다 74% 정도로 가장 높은 효율 향상을 보여주었다. 실제로 최적 예측조건에서 실증실험을 수행한 결과 본 모델의 타당성이 입증되었다. 본 연구에서 얻은 섬유소 분해 효소 생산 조건 최적화에 관련된 자료들은 산업적으로 효소를 생산하고자 할 때 설계에 유용하게 사용될 것이다.

In an effort to isolate novel strains expressing a thermostable esterase that hydrolyzed the rac-ketoprofen ethyl ester to ketoprofen in the stereospecific manner, we screened various soils and composts from broad ecological niches in which the activity was expected to be found. Three hundreds of microbial strains were tested to determine their ester-hydrolyzing activity by using an agar plate containing insoluble tributyrin as an indicative substrate, and then further screened by activity on the (R,S)-ketoprofen ethyl ester. Twenty-six strains were screened primarily at high growth and incubation temperature and further compared the ability to ethyl ester-hydrolyzing activity in terms of conversion yield and chiral specificity. Consequently, a strain JYl44 was isolated as a novel strain that produced a (R)-stereospecific esterase with high stability and systematically identified as a Bacillus stearothermophilus JY144. The enzyme indeed stables at a broad range of temperature, upto 65 $^{\circ}C$, and pH ranging from 6.0 to 10.0. The optimal temperature and pH for enzymatic conversion were 50 $^{\circ}C$ and 9.0, respectively. Based on the observations that resulted a poor cell growth, and enzyme expression in wild type strain, we further attempted the gene cloning into a general host Escherichia coli and determined its primary structure, concomitantly resulting a high level expression of the enzyme. The cloned gene had an open reading frame (250 amino acids) with a calculated molecular mass of 27.4 kDa, and its primary structure showed a relative high homology (45-52 %) to the esterases from Streptomyces and Bacillus strains. The recombinant whole cell enzyme could efficiently convert the rac-ketoprofen ethyl ester to (R)-ketoprofen, with optical purity of 99 % and yield of 49 %.

화학독립영양미생물 Aeromonas sp. strain JS-l는 호수와 웅덩이의 표층수에서 분리 동정 되었고 분리된 strain JS-1은 에너지원과 탄소원으로써 각각 $H_2$와 $CO_2$를 이용하였다. RubisCO(EC 4.1.1.39)는 Aeromonas sp. strain JS-l으로부터 ammonium sulfate 침전과 DEAE-sepharose CL-6B, gel filtration chromatography 방법으로 정제되었다. RubisCO의 분자량은 gel filtration에 의해 대략 560 kDa임을 확인되었으며, SDS-PAGE에 의해 Large subunit(56 kDa)와 Small subunit(14 kDa)로 구성된 $L_8S_8$구조를 가지고 있음이 확인되었다. Ribulose 1,5-bisphosphate (RuBP), $NaH^{14}CO_3$ 와 $Mg^{++}$의 Km값은 각각 0.25 mM, 5.2 mM, 0.91 mM이었으며, 효소반응의 최적온도는 $50^{\circ}C$였으며, 열 안정성은 $45^{\circ}C$까지 안정하였다.

부패한 사과로부터 bacterial cellulose (BC)를 생산할 수 있는 균주를 분리한 후 배양조건에 따른 BC의 생산량을 조사한 결과 BC의 생산량은 진탕배양한 경우가 정치배양한 경우보다 약 1.5배 높았다. BC의 생산량을 높이고자 mutagen으로 UV와 cylcloheximide를 사용함으로써 BC 생산량을 약 3배 증가시킬 수 있었다.. 미생물에 의해 생성된 BC는 종이나 펄프와는 달리 pectin, 납, 유지, 단백질, 무기질 등의 불순물을 함유하지 않는 filter paper와 성질이 유사한 것으로 나타났다.

균체 생산성 실험과 chitinase 생산성 실험을 비교해 볼 때, chitinase만을 생산하는 조건 에서는 배지성분에 chitin을 첨가해 주는 것이 좋으나, 해충 방제용으로 살균력을 증진시키기 위하여 균체량과 chitinase의 생성량 및 산업적, 경제적 사용이 용이한 배지를 고려할 때에는 쌀겨와 밀기울이 첨가된 배지가 좋은 배지임을 알 수 있었다. 또한 이 배지를 이용하였을 경우 균체는 1X$10^8$ cfu/g, chitinase는 370mU/g로 생산되었으며 생물검정결과 53-64%의 탁월한 살충효과를 확인 할 수 있었다.

A lignin degradation bacteria, symbiotic bacteria was isolated from the gut of Sympetrum depressiusculum and tested for its lignin degrading activity using lignin model compounds and related aromatic compounds. The strain was identified as Serratia marcescens HY-5 based on the 165 rDNA, cellular fatty acid composition, biochemical and physiological characteristics. S. marcescens showed 40-50% lignin degrading activity in the media that contained vaillin, guaiacol and dealkaline lignin. S. marcescens showed three ligninase activities [Jaccase, lignin peroxidase(LiP) and Manganase peroxidase(MnP)]. Addition of dealkaline lignin to the basal media increased about 6fold of laccase activity. Vanillic acid or vanillin increase 1.3fold of MnP activity and p-coumaric acid increased 12fold of LiP activity which added to the basal medium.

A thermostable xylanase from Thermotoga maritima (Xyn B) cleaves several pNP-glycosides of monosaccharides. We found that the initial product of the cleavage of pNP-xyloside (pNP-Xy1) was a disaccharide, not xylose, indicating that xylosyl unit of pNP-Xyl was transglycosylated to another pNP-Xyl. We determined that the disaccharide was xylobiose which has the linkage of the ${\beta}$ 1-4, and described the reaction mechanism of the Xyn B. Also, we produced the several pNP-glycosides and propyl-disaccharides from the transglycosylation of Xyn B with varial glycosides and/or 1-propanol. All reaction products were purified by column chromatography (Toyo-pearl HW-40C, 45 cm${\times}$2.5 cm or 45 cm ${\times}$ 2.5 cm${\times}$ 2). The isolated products were analyzed by means of 1D and 2D NMR.

재접힘 고체상 재접힘은 높은 재현성을 보였으며 고체상 재접힘된 단백질은 Native와 같은 구조를 형성하였다. 따라서 이 연구는 고체상 재접힘 방법이 분자간의 상호작용을 억제하는 것이 응집현상을 탈피하게된 결과 일 것이라는 것에 의해 재접힘 수율을 높일 수 있다고 기대한다.

We have performed batch chromatography experiments to calculate parameters which are used to design SMB chromatography system. Isotherms of S- and R-ketoprofen enantiomers were gained from small amount loading experiments in a column, and flow rate of four zones of SMB chromatography were calculated using mass balance equations. As a results, diameter and length of columns were 10mm and 600mm, and flow rate of each zone were as follows; $Q_{Feed}$=1.00ml/min ,$Q_{Elu}$=2.81ml/min ,$Q_{Raf}$=1.81ml/min ,$Q_{Ext}$=2.00ml/min , and $Q_{Rec}^{TMB}$=10.75ml/min , From the Darcy's equation, the pressure drop in whole columns of SMB chromatography was 128bar.

용균작용을 하는 lysozyme을 난백으로부터 분리하기 위해 이온교환크로마토그래피를 사용하였다. 용출시에 gradient를 걸어준 결과 젤과 약하게 결합된 단백질과 강하게 결합된 단백질이 분리되어 나옴을 SDS-PAGE와 Lowry methode를 통하여 알 수 있었다.

In order to improve the physical properties of food such as texture and food self-life. Transglutaminase(mTG) from Streptoverticillum morbarense was prepared. In the preliminary experiments, presence of proteases in the crude enzyme did not improve the texture of dough, which mean the inteference of mTG reaction by the proteases. Among the cation exchange resins tested for the removal of proteases, Monoplus S 100(Bayer, Germany) was the most efficient resin with 20 fold increase in the mTG/protease activity ratio. By further purification steps with a quaternary ammonia salt resin and a gel permeation chromatography, proteases were effectively removed from the preparation. Therefore, the improvement of flour texture was shown by adding the protease-free mTG.

A separation/purification process for enzymatic sugar ester production was investigated The crude reaction mixture contained sugar ester and unreacted fatty acid in acetone. The reaction mixture was mixed with KOH/phosphate buffer. Hexane was then added to enhance phase separation. Three phases formed: a lower aqueous phase containing nothing of interest, a polar organic solvent middle phase that contained mostly fatty acid soaps, and a hexane-rich upper phase that contained mostly sugar ester. Distribution coefficient of each component was measured.

Iron is an essential nutrient for most organisms, which supplied to them in a protein-iron complex known as ferritin. Ferritins are multimeric proteins those are consisted of spherical shell of 24 subunits defining a cavity of about 8nm in diameter. Soluble form of ferritin was separated from disrupted cells, followed by silica powder adsorption. Ferritin was recovered from silica-poweder by distiiled water, which was applied to DEAE anion exchage chromatography. Collected fractions from the DEAE column were assayed to gain the amount and the purity of ferritin by using GF-HPLC.

The method of foam fractionation can be applied to enrich proteins from a dilute protein solution if the proteins are hydrophobic and foam. If a protein, such as invertase, is hydrophilic, a dilute solution containing this protein may not foam. In that case, a batch foam fractionation process may not be appropriate for recovering a concentrated solution of that protein. In this paper, various concentrations of invertase were added to a dilute solution containing bromelain (a hydrophobic protein), in order to determine how the presence of a hydrophilic protein can affect the recovery of the desired hydrophobic protein. The effect of invertase on bromelain recovery was studied here at an initial bulk solution pH of 5 and an air superficial velocity of 4.6 cm/s.

인간 fibroblasts의 receptor를 통한 LDL의 섭취와 분해에 대하여 보다 개선되어진 수학적/동역학적 모델을 제시하였다. 관련된 동역학적 모델의 hierarchy를 통하여 세포 멤브레인 표면으로 recycle되는 receptor의 선택적 insertion 정도를 나타내는 파라미터, ${\alpha}$를 가지는 모델을 제안하였다. 여러 가지의 LDL 농도의 미디움과 여러 가지의 실험조건에서 모델예측을 수행하였는데, Brown과 Goldstein의 많은 실험데이터에 잘 일치하였다.

Xanthmonas oryzae MGL21유래의 CFTase의 repeat영역을 deletion 시킨 ${\triangle}N{\triangle}C$ deletion mutant는 protein 정제결과 약 90kDa 이있으며 pH6.5, $45^{\circ}C$에서 최적 효소 반응을 하였다. 또한 Inulin과 반응시켰을 때 CFTase는 main product가 CF인데 비해 ${\triangle}N{\triangle}C$ deletion mutant는 main product가 fructooligosaccharide였다. 이러한 결과로부터 CFTase의 N말단 repeat영역과 C말단 repeat영역을 제거하였을 경우 endoinulinase와 활성이 유사하며, 유전자 크기 및 아미노산 서열도 유사함을 알 수 있었다.

본 연구에서는 기능성 식품 첨가물 소재 또는 의약품으로 사용 가능한 한천올리고당의 대량생산을 위하여 agarase 생산균주인 Bacillus cereus ASK202에 대해 유전자 cloning 방법을 이용하여 한천분해효소 고생산성 균주로의 개발을 시도하였다. 원균주의 chromosomal DNA를 무작위적으로 절단하여 agarase를 생산하는 gene 부위를 선별한 결과, 83,300 Da의 agarase(Eba1)를 생산하는 재조합 균주 E. coli BL21(DE3)/pEBA1를 얻을 수 있었다. E. coli BL21(DE3)/pEBAl에서 유도물질로 IPTG를 첨가한 후 induction 5시간 후에 agarase가 원균주에 비해 8배 증가한 양이 고발현되었다. 발현된 agarase는 Asx(Aspartic acid, Asparagine), Glycine와 같은 산성 아미노산의 함량과 Alanine과 같은 중성 아미노산의 함량이 높았으며, pH 5.6, $40^{\circ}C$에서 최적의 활성을 나타내었다. 최적 기질 agar에 대한 $K_m$과 $V_{max}$값은 0.068 mg/$m{\ell}$과 0.094mg/$m{\ell}{\cdot}min$으로 한천의 ${\beta}$-결합을 자르는 ${\beta}$-agarase로 판명되었다.

We propose new encoding method for numerical data in DNA using temperature gradient. To represent numerical values in DNA sequences, we introduce melting temperature. Since DNA strands representing smaller values have a lower Tm, they tend to denature with ease and also easily amplified by denaturation temperature gradient PCR. We also implement a local search molecular algorithm using temperature gradient, which is contrasted to conventional exhaustive search molecular algorithms. The proposed methods are verified by solving an instance of the travelling salesman problem. We could effectively amplify the correct solutions and the use of temperature gradient made the detection of solutions easier.

DNA vaccines use eukaryote expression vectors to produce immunizing proteins in the vaccinated host and it represents a novel approach to vaccine and immuno-therapeutic development. We constructed a 2.9 kb compact plasmid vector (pVAC) which contains CMV promoter, polycloning site, BGH poly A terminator, ampicillin resistance gene and PBR322 origin. Enriched unmathlyated CpG motifs have introduced into pVAC-ISS1 and pVAC-ISS2 which are derived from pVAC for enhancing Thl responses. These plasmid DNAs rapidly induced interleukin 6 secretion in vivo. It is expected that these vectors will contribute to the DNA inoculation against infectious disease and various cancers without adjuvant.

Antioxidative noudle products were developed with mixed material and antioxidative liquid extracted from natural products. Matrrials were mixed with wheat flour, rice flour, starch and green tea. 3% of green tea was suitable mixing ratio from the mastication data of texture meter, and proper water contents were 33% to prepare these noodles. The suitable mixing ratio of materials were wheat flour(85%), starch( 12%) and greentea(3%) from the physical data of texture meter. Since AUC value of antioxidative liquid was more higher than that of tap water, antioxidative noodle products were expected for good health and preservation of some diseases.

In this study, a method for the localization of pregnancy-specific proteins from cow urine on 2-D gel have been established. The proteins were digested with trypsin in gel and then analyzed with MALDI- TOF or transferred to a membrane and microsequenced. To examine the pregnancy associated protein spot 2 as a diagnosis marker in bovine urine 2-D western blotting was performed. This antibody was reacted specifically in the protein of pregnant cow’s urine. Consequently spot 2 was identified and found to be good candidates for developing cow pregnancy detection assay kit.

The esterification reaction of previously obtained l,4-sorbitan with acrylic acid using Novozym 435 was carried out in t-butanol as solvent. Immobilized lipase Novozym 435 showed high enzymatic activity at $50^{\circ}C$ in t-butanol and optimum contents of Novozym 435 added in the esterification reaction was 3%(w/v). The maximum conversion rate was 55.8% when initial concentration was 50g/L and conversion rate of this reaction was 63.5% when the molar ratio of l,4-sorbitan to acrylic acid was 1:3.

Individual surface(hydrophilic/hydrophobic) were prepared and mammalian cells were cultured on the hydrophilic region. For drug test, cancer and normal cells were treated with Taxol, as an example. Our system was compared with MTT assay. CHO cells were resistant to Taxol up to 100 nM in both Methods. However, A549 cells was sensitive at 100 nM Taxol in the 2 day-treatment. Cervical carcinoma cell, HeLa, was very sensitive to Taxol. In our system, the cells were not shown from above 20 nM Taxol treatment. Our system was competitive to MTT assay in animal cells for drug test.