Fermentation characteristics of recombinant Saccharomyces cerevisiae containing the xylose reductase gene from Pichia stipitis were analyzed in an attempt to convert xylose to xylitol, a natural five-carbon sugar alcohol used as a sweetener. Xylitol was produced with a maximum yield of 0.95 (g xylitol/g xylose consumed) in the presence of glucose that is used as a cosubstrate for cofactor regeneration. However addition of glucose caused inhibition of xylose transport and accumulation of ethanol. Such problems were solved by adopting glucose-limlted fed-batch fermentation. This process done with S, cerevisiae EHl3.15:pY2XR at$30\;^{\circ}C$ resulted in 105.2g/L xylitol concentration with maximum productivity of 1.69 g $L^{-1}$ $hr^{-1}$.

A. pullulan ATCC 42023를 사용하여 고분자량의 pullulan을 대량생산하기 위한 연구로써, 탄소원의 농도, 용존산소량과 pH가 균체 성장 및 pullulan 생산에 미치는 영향에 대하여 연구하였다. Glucose의 당 농도에 따른 실험결과에서 glucose 농도가 0.3M (54g/1)일 때 최대의 pullulan 생산률을 나타내었고, 0.3M (54g/1)이상에서는 productivity가 급격하게 감소하는 기질 저해현상을 나타내었다. 삼각플라스크를 이용한 초기 pH의 영향에 대한 실험결과, 초기 pH 6.5는 $11.98g/{\ell}$의 pullulan을 생산하였고, pH 4.5로 일정하게 조절한 결과, pullulan 생산은 $13.31g/{\ell}$의 최대값을 나타내었다. 그러나 균체성장은 pH 6.5에서 가장 높았다. 그리고 용존산소량의 증가로 pullulan productivity를 증가시킬 수 있었다. 또한 산업적으로 유용한 고분자량의 pullulan을 생산하기 위한 탄소원으로 아미노당인 glucosamine을 사용하여 pullulan의 생성 및 분자구조 변형에 대하여 실험하였다. 탄소원 농도 50g/1 기준에서, glucose와 glucosamine을 40 : 60의 비율로 혼합하여 공급 하였을 때 분자량 20만 이상의 mediun size 및 200만 이상의 high molecular weight의 productivity가 가장 높았으며, A. pullulans의 specific growth rate도 가장 높았다.

생물공정의 운전에 있어서 적절한 공정변수가 부족한 경우가 많다. 이것은 멸균과정을 견딜 수 있는 신뢰성 높은 센서가 부족하기 때문이다[1]. 생물공정에 주로 사용되는 센서로서는 온도, pH, D.O., rpm, viscosoty 등이 있으나 이 센서들은 배양액의 물리적 혹은 화학적 상태를 측정할 수 있는 경우가 대부분이다[2]. 미생물의 대사활동과 관련이 있는 공정 변수로는 배출가스의 성분을 측정하여 얻을 수 있는 Oxygen uptake rate, Carbon dioxide evolution rate 및 Respiratory quotient가 있으며 현재 생물공정의 운전에 사용되고 있다[3]. 그러나 반복적인 센서의 보정과 연결관의 잦은 청소 및 보수를 필요로 하여 제한적으로 사용되고있는 실정이다. 자동화된 습식분석장치, Gas chromatograph, High Performace Liquid Chromatograph 혹은 Mass spectrophtometry 등을 온라인 샘플 처리장치와 연결하여 발효조의 배양액의 성분을 온라인으로 분석하고 공정의 운전에 응용하는 사례가 많이 발표되었다[4-6]. 고가의 장비 및 운전의 번거러움이나 추가적인 인력이 필요하므로 역시 특별한 경우에만 사용되고 있다. 이외에도 여러 종류의 온라인 센서 및 바이오 센서등이 개발되어 사용되고 있으나 역시 그 사용범위는 특수한 영역에 한정되어있다. 이와 같이 새로운 센서를 개발하여 공정변수를 측정하려는 시도중의 하나가 소프트웨어 센서의 개발이다. 이 것은 공정상에서 발생하는 1차 공정변수를 이용하여 배양액의 상태 혹은 2차적인 공정 변수를 추측해내는 것이다. 대부분의 경우 기존의 공정 변수를 사용하므로 추가적인 비용이 들지 않고 소프트웨어의 형태로 구현되므로 센서의 보정과 설치 및 유지관리의 노력이 매우 적은 장점이 있다. 본 연구에서는 생물공정에서 자동제어 과정에서 발생하는 여러 가지 공정상의 제어 신호로부터 새로운 공정 변수를 얻어내고자 시도하였다. 대부분의 생물공정에서는 pH의 자동제어가 필수적인데 자동제어 과정에서 발생하는 pH 제어 신호 및 pH의 변화 응답신호를 이용하여 배지의 완충용량의 변화와 알칼리의 소비속도를 온라인으로 측정할 수 있었다. 여기에 인공지능망을 설계하여 균체의 량을 온라인으로 추정하는 방법을 개발하였다 [7].산업용 발효조의 운전 온도는 주로 냉각수의 단속적인 공급에 의하여 항상 일정하게 조절된다. 따라서 냉각수의 냉각량을 측정하면 미생물의 배양시 발생하는 대사열량을 측정할 수 있게 된다. 본 연구에서는 실험실의 발효조를 냉각수의 단속적인 공급에 의하여 자동온도 조절이 되도록 개조하고 여기에 냉각수의 유출입 지점에 온도센서를 부착하여 냉각수의 온도를 측정하고 냉각수의 공급량과 대기의 온도 등을 측정하여 대사열의 발생을 추정할 수 있었다. 동시에 이를 이용하여 유가배양시 기질을 공급하는 공정변수로 사용하였다 [8]. 생물학적인 폐수처리장치인 활성 슬러지법에서 미생물의 활성을 측정하는 방법은 아직 그다지 개발되어있지 않다. 본 연구에서는 슬러지의 주 구성원이 미생물인 점에 착안하여 침전시 슬러지층과 상등액의 온도차를 측정하여 대사열량의 발생량을 측정하고 슬러지의 활성을 측정할 수 있는 방법을 개발하였다.

효모를 이용하여 높은 생산성으로 고농도의 erythritol을 생산하기 위하여 ‘세포 성장단계’와 ‘erythritol 생성단계’로 나누어 2 단계 유가식 배양을 수행하였다. 세포 성장 단계에서는 pH-stat 유가식 배양을 수행하여 3.62 g/L-H의 생산성으로 76 g/L의 균체 농도를 얻을 수 있었다. pH-stat 유가식 배양을 수행하여 고농도의 균체 농도를 얻은 후에 멸균되지 않은 분말형태의 glucose를 발효조에 400 g/L가 되도록 공급하여 세포에 osmotic stress를 가해준 결과 41%의 수율로 187 g/L의 erythritol을 생산할 수 있었으며, 이때의 생산성은 2.79 g/L-h로서 기존에 보고된 어느 결과보다도 높은 값을 얻을 수 있었다. Production phase에서 erythritol 생산에 대한 삼투압의 영향을 알아 본 결과 glucose의 농도가 증가할 수록 생성되는 erythritol 농도가 증가하였으며 세포 농도에 따른 erythritol 생산 수율은 큰 차이는 없었으나 세포 농도가 증가할수록 이에 비례하여 erythritol 생산성이 크게 증가하였다. Erythritol 생산에서 용존 산소량에 관계없이 butyric acid가 생성되었으며 40%의 DO 미만에서는 citric acid가 생성되고, 40% 이상의 DO에서는 gluconic acid가 생성되어 용존 산소량에 의하여 metabolic shift가 유도되었다. 고 농도의 glucose 첨가 후 용존 산소량을 20%로 조절하면서 pH를 3.0으로 낮춘 결과 citrate synthetase의 활성이 억제되어 citric acid가 생성되지 않았으며 gluconic acid 농도는 $KH_2PO_4$에 의하여 증가하고 $MgSO_4{\cdot}7H_2O$ 또는 $(NH_4)_2SO_4$에 의하여 감소되었다. Butyric acid 농도는 $KH_2PO_4$ 또는 $(NH_4)_2SO_4$에 의하여 감소되었으며 synergic effect에 의하여 억제되었다

Production of dye-decolorizing enzyme was investigated with the cultivation of Geotrichum candidum Dec1 (abbreviated to Dec1) using molasses as a cheap raw materials. Molasses was found to stimulate the enzyme production as well as a role of carbon source, because the production increased with molasses content up to 50 g/L. However, the severe inhibition of dye-decolorizing enzyme activity was observed even at low concentration of molasses 10 g/L when purified decolorizing peroxidase was used. Its inhibitory effect was reduced through the cultivation of Dec1. The fractions of molasses separated by a gel chromatography showed the different degrees of inhibition. As a way to reduce the inhibitory effect, the dilution of culture broth was examined, and the total decolorizing activity for Reactive blue 5 increased 7 times as much as that of original culture broth by 30 times dilution. On the basis of result, we proposed a process scheme which can fully utilize both positive and negative effect of molasses in dye-decolorizing process using molasses.

식물배양세포의 항산화기구 이해를 위하여 다양한 식물종에서 유도된 100종의 배양세포주를 대상으로 항산화효소 활성 및 저분자항산화물질을 분석하였다. 배양세포의 SOD와 POD 활성은 식물체에 비해 매우 높았으며, 특히 고구마와 카사바 배양세포는 각각 POD와 SOD 활성이 가장 높아, 항산화효소의 생산 및 항산화기구 해석에 좋은 소재임이 시사되었다. 배양세포의 평균 ascorbate 함량은 식물체에 비해 1/100 수준으로 낮았으며, glutathione 함량은 배양세포가 식물체보다 약 2-3배 높았다. 흥미롭게도 ascorbate와 glutathione의 환원형과 산화형의 비율은 식물종에 따라 큰 차이가 있음이 밝혀졌다. 결론적으로 식물배양세포는 높은 산화적인 스트레스에서 배양되는 것이 생화학적으로 확인되어 항산화물질 생산 및 항산화기구 연구에 좋은 소재임이 시사되었다.

유용약용식물의 배발생세포를 MS (murashige and Skoog) 액체배지에서 배양하여 균일하게 대량으로 증식시키고, 이들이 체세포배과정을 거처 유식물체로 발달시키는 기술을 개발하였다. 이 방법에 의해 일부 귀중한 약용식물은 공장생산이 가능하게 되었고, 싼가격에 약용식물의 유용성분을 추출하여 건강식품, 음료, 차, 의약품 및 화장품의 원료로 사용할 수 있다.

Polynuclear aromatic hydrocarbon (PAH) compounds are highly carcinogenic chemicals and common groundwater contaminants that are observed to persist in soils. The adherence and slow release of PAHs in soil is an obstacle to remediation and complicates the assessment of cleanup standards and risks. Biological degradation of PAHs in soil has been an area of active research because biological treatment may be less costly than conventional pumping technologies or excavation and thermal treatment. Biological degradation also offers the advantage to transform PAHs into non-toxic products such as biomass and carbon dioxide. Ample evidence exists for aerobic biodegradation of PAHs and many bacteria capable of degrading PAHs have been isolated and characterized. However, the microbial degradation of PAHs in sediments is impaired due to the anaerobic conditions that result from the typically high oxygen demand of the organic material present in the soil, the low solubility of oxygen in water, and the slow mass transfer of oxygen from overlying water to the soil environment. For these reasons, anaerobic microbial degradation technologies could help alleviate sediment PAH contamination and offer significant advantages for cost-efficient in-situ treatment. But very little is known about the potential for anaerobic degradation of PAHs in field soils. The objectives of this research were to assess: (1) the potential for biodegradation of PAH in field aged soils under denitrification conditions, (2) to assess the potential for biodegradation of naphthalene in soil microcosms under denitrifying conditions, and (3) to assess for the existence of microorganisms in field sediments capable of degrading naphthalene via denitrification. Two kinds of soils were used in this research: Harbor Point sediment (HPS-2) and Milwaukee Harbor sediment (MHS). Results presented in this seminar indicate possible degradation of PAHs in soil under denitrifying conditions. During the two months of anaerobic degradation, total PAH removal was modest probably due to both the low availability of the PAHs and competition with other more easily degradable sources of carbon in the sediments. For both Harbor Point sediment (HPS-2) and Milwaukee Harbor sediment (MHS), PAH reduction was confined to 3- and 4-ring PAHs. Comparing PAH reductions during two months of aerobic and anaerobic biotreatment of MHS, it was found that extent of PAHreduction for anaerobic treatment was compatible with that for aerobic treatment. Interestingly, removal of PAHs from sediment particle classes (by size and density) followed similar trends for aerobic and anaerobic treatment of MHS. The majority of the PAHs removed during biotreatment came from the clay/silt fraction. In an earlier study it was shown that PAHs associated with the clay/silt fraction in MHS were more available than PAHs associated with coal-derived fraction. Therefore, although total PAH reductions were small, the removal of PAHs from the more easily available sediment fraction (clay/silt) may result in a significant environmental benefit owing to a reduction in total PAH bioavailability. By using naphthalene as a model PAH compound, biodegradation of naphthalene under denitrifying condition was assessed in microcosms containing MHS. Naphthalene spiked into MHS was degraded below detection limit within 20 days with the accompanying reduction of nitrate. With repeated addition of naphthalene and nitrate, naphthalene degradation under nitrate reducing conditions was stable over one month. Nitrite, one of the intermediates of denitrification was detected during the incubation. Also the denitrification activity of the enrichment culture from MHS slurries was verified by monitoring the production of nitrogen gas in solid fluorescence denitrification medium. Microorganisms capable of degrading naphthalene via denitrification were isolated from this enrichment culture.

The Internet can be considered as a good environment for dwelling of many types of virtual lives. If we could build up the proper ecosystem with the lives in the internet, It would be possible to implement the system as a life. The life system thus obtained would provide us some decisive advantages, which is hardly available by the reduction paradigm. In this paper, we presented an ecosystem based on Internet, which will be followed by the discussion of its meaning in terms of usefulness.

Enhanced biological phosphorus removal (EBPR) is not always successfully achieved by anaerobic/aerobic operation. It has been reported that the EBPR deterioration was caused by the outgrowth of glycogen-accumulating organisms (GAO) over polyphosphate-accumulating organisms (PAO). It was found that pHcould be a tool which might induce the success of EBPR in a sequencing batch reactor (SBR) supplied with acetate. When the pH of anaerobic phase was controlled at 7.0, the operation resulted in failure of EBPR. However, when the pH of anaerobic phase increased up to 8.4, complete EBPR was achieved. We explained the mechanism of pH effect on the competition between GAO and PAO with experimental results and previously proposed biochemical models.

Long chain fatty acids (LCFA) are potential inhibitors of bacteria involved in anaerobic digestion because of their surface activity. Precipitation of long-chain fatty acids with iron can improve the anaerobic degradation due to their precipitation and reducing surface properties. Degradation of stearic acid was improved in the presence of iron (II). The methane production was increased 1.6 times as compared to control. Iron-containing soil was applied for degradation of vegetable oil as model case. The methane production was increased 1.5 times as compared to control. Yield of methane production was 0.09 and 0.06L/g COD in experiment and control respectively. Optimum COD/Fe ratio was found 20 mg/mg. Iron (II) can be produced in the treatment system from iron (III) hydroxide or iron containing minerals.

Hexavalent chromium was removed by means of biosorption onto the protonated brown seaweed biomass. During the biosorption Cr(VI) was reduced to Cr(III), which resulted in accumulation of Cr(III) in the solution. The Cr(VI) reduction rate increased with increases of initial Cr(VI) and biosorbent concentrations and decrease of solution pH. Based upon the experimental results at various conditions, we suggested the mechanism for the chromium removal as following serial reactions: (1) sorption of anionic Cr(VI) onto the positively charged site of biomass, (2) reduction of Cr(VI) to Cr(III) on the positively charged site, (3) desorption of Cr(III) from the positively charged site, and (4) sorption of cationic Cr(III) onto the negatively charged site of biomass.

본 연구는 4가지 종류의 재조합 발광성 미생물을 이용하여 내분비계 장애물질로 알려진 여러 가지 물질에 대한 cellular toxicity를 유전자 손상, 단백질 손상, 산화적 손상, 생물막 손상으로 구별하여 확인하였다. 4가지 발광성 미생물의 반응성에 따라 내분비계 장애물질의 독성 형태를 규명할 수 있었고, 생명체에 미치는 독성 정도를 확인할 수 있었다. 또한 유전자 손상을 탐지할 수 있는 DPD2794의 경우 유전자 손상을 일으키는 형태에 따라 두 그룹으로 보다 세분화가 가능하였다. 따라서 본 연구결과를 바탕으로 내분비계 장애물질이 호르몬 교란으로 인한 피해뿐만 아니라 cellular toxicity로 인한 피해 역시 입힐 수 있는 것으로 확인하였고, 이들 발광성 박테리아를 이용하여 그 독성 형태를 정확하게 파악할 수 없었던, 유해 물질들의 분류를 위한 screening method로의 개발 역시 가능할 것이다.

본 연구의 목적은 식품부산물로부터 유기산 생산 및 분리 농축에 있다. 회분식 발효를 실시하여 균주의 최적 발효조건을 구하여 세포 재순환식 연속 발효공정에 적용한 결과, CSL의 최적 함량은 2.5%로 나타났으며 수율과 생산성은 각각 0.80g total acid/g glucose, 0.26g total acid/L/h로 향상 되었다. 세포 재순환식 연속발효 결과, 최대 유기산 생산성은 희석률 0.2/h에서 3.32g organic acid/L/h로 회분식 발효와 비교해 볼 때 최대 생산성으로는 13배, 세포량으로는 22배 증가한 것으로 나타났다. 또한, 최적 추출시스템은 30%w/w TOA/MIBK로 나타났으며, 발효액내의 유기산을 약 90%까지 분리 및 농축할 수 있음을 확인하였다.

본 연구에서는 순환식 돈분 폐수 처리 시스템에서의 미생물 분포에 따른 폐수 처리 효과를 모델링하기 위해 신경회로망과 PCA를 이용하는 새로운 방법을 제안하였다. PCA 분석 결과를 바탕으로 신경회로망의 최적 입력 조건을 찾고, 실측 데이터를 이용하여, 폐수 처리 시스템의 각 탱크를 별도로 학습함으로써 비교적 적은 수의 데이터에도 불구하고 정확한 모델링 결과를 얻었다. 제안한 시스템은 폐수 처리 시스템의 효과적인모니터링 시스템으로 사용할 수 있으며, 향후 실제 돈분 처리 시스템에서 원하는 기준의 방류수를 얻기 위한 최적의 입력조건 (미생물밀도 등)을 결정하는데 있어서 에뮬레이터로 사용될 수 있을 것으로 기대된다.

유기산 생산을 위한 Propionibacterium acidipropionici의 발효 특성을 규명하기 위하여 ATCC 4965, 4875, 25562의 세 균주를 선별하였다. 세포 성장에 따른 발효 실험에서는 Peptone, Yeast extract, pH별로 실시되었으며, 각각 1.5 %, 0.75%, $5.5{\sim}7.5$에서 유기산 생산성이 최적으로 나타났다. 회분식 발효결과 세 균주 모두 pH 6.0에서 유기산 수율과 생산성이 pH 7.0에 비해 더 높았으며, 그 중에서도 P.acidipropionci ATCC4965균주가 유기산 생산성이 0.29g total acids/L/h로 가장 우수한 것으로 나타났다.

In recent years, hybrid neural network approaches which combine neural networks and mechanistic models have been gaining considerable interests. These approaches are potentially very efficient to obtain more accurate predictions of process dynamics by combining mechanistic and neural models in such a way that the neural network model properly captures unknown and nonlinear parts of the mechanistic model. In this work, such an approach was applied in the modeling of a full-scale coke wastewater treatment process. First, a simplified mechanistic model was developed based on the Activated Sludge Model No.1 and the specific process knowledge, Then neural network was incorporated with the mechanistic model to compensate the errors between the mechanistic model and the process data. Simulation and actual process data showed that the hybrid modeling approach could predict accurate process dynamics of industrial wastewater treatment plant. The promising results indicated that the hybrid modeling approach could be a useful tool for accurate and cost-effective modeling of biochemical processes.

Marine microorganism strain 96CJ10356 produced exopolysaccharides, designated as EPS-R. To optimize culture conditions for the production of EPS-R, carbon, nitrogen, mineral salt, temperature, and pH were examined. STN medium was suggested as follow; sucrose 20, tryptone 10, NaCl 10, $MgSO_4$ 5, $CaCl_2$ 1, $KH_2PO_4$ 0.0076, $K_2HPO_4$ 0.0083, $FeCl_2$ 0.005, $MnCl_2$ 0.001, $NaMoO_4$ 0.001, $ZnCl_2$ 0.001 (g/1) and pH 7.0 About 9.23 g/l of EPS-R was obtained from the STN medium after cultivation for 120 h at $25^{\circ}C$ in 5-liter jar fermentor with an aeration rate of 0.17 vvm. Apparent viscosity and flocculation activity of the culture broth were increased with the production of the EPS-R and the maximal values were reached to 415 cp and 1400 units/ml against 0.5 % activated carbon, respectively.

Highly open porous polymer matrices are required for high density cell seeding, efficient nutrient, and oxygen supply to the cells cultured in the three dimensional matrices. However, there are severe problems of mass transfer limitations within the cell/scaffolds culture system. Thus we hypothesize that continuos-flow culture conditioning of cells with the scaffolds may improve the cell viability and the differentiated function. In this study, we fabricated porous PLGA scaffolds by using gas-foaming/salt-leaching method as previous described. Viscous PLGA gel paste contains ammonium bicarbonate particulates, acting as a gas-foaming agent as well as a salt-leaching porogen, were cast into Teflon mold and dried. Ammonium bicarbonate salt upon contact to an acidic aqueous solution evloves gaseous ammonia and carbon dioxide by itself. And we conjugated galactose moiety [AGA; $N-(aminobuty1)-O-{\beta}-D-galactopyranosyl-(1{\rightarrow}4)-D-glucoamide]$ to the terminal end group of a PLGA to increase the cell adhesion and matain the differentiated function of hepatocytes. Cell-seeded scaffolds were secured in a flow bioreactor chamber and exposed to continuous flow at 5 ml/min. As a result of our study, the high yield of hepatocytes attachment was accomplished by increasing the concentration of PLGA-AGA conjugate in polymer scaffolds and cells in the scaffolds under continuos flow condition maintained a high level of viability and albumin secretion rate of cultured hepatocytes showed a higher level that of control groups.

Large-scale resolution of epoxides by the yeast Rhodotorula glutinis was demonstrated in an aqueous/organic two-phase cascade membrane bioreactor. Due to the chemical instability and low solubility of epoxides in aqueous phases, an organic solvent was introduced into the reaction mixture in order to enhance resolution of epoxide. A cascade hollow-fiber membrane bioreactor was used (i) to minimize the toxicity of organic solvents towards the epoxide hydrolase of Rhodotorula glutinis, and (ii) to remove inhibitory amounts of formed diol from the yeast cell containing aqueous phase. Dodecane was selected as a suitable solvent and 1,2-epoxyhexane as a model substrate. By use of this membrane bioreactor, highly concentrated (0.9 M in dodecane) enantiopure (>98% ee) (S)-1,2-epoxyhexane (6.5 g, 30% yield) was obtained from its racemic mixture.

GFP는 박막상태에서 쉽게 전자소자로 제작할 수 있으며, 제작된 전자소자는 빛의 여부에 따라 흐르는 전류의 조절이 가능한 광다이오드 특성을 나타내는 것으로 확인되었다. 또한 박막상태에서의 일방향 전자전달 속도와 전자전달 메커니즘이 확인되어, 향후 분자전자소자의 제작시 기반기술로 제공될 수 있을 것으로 사료된다.

1. A. ficuum의 genomic library 검색을 통해 Axe 유전자를 포함하고 있는 5.0 kb의 XbaI DNA 절편을 cloning 했다. Cloning 된 절편의 부분 염기서열 결정 결과 약 1.4 kb의 AXE coding 부위를 확인했으며, cDNA cloning과 그 염기서열의 결정을 통해 AXE coding 부위 내에는 두 개의 intron 이 존재함이 확인되었다. 2. AXE coding 부위의 아미노산 잔기 서열 검색 결과 A. awamori의 AXE와 약 92%의 상동성과 95%의 유사성이 있음이 확인 되었다. 3. 약 900 kb의 AXE의 cDNA를 yeast의 YEp352 vector의 GAL1 promoter의 전사 방향으로 cloning한 후 발현시킨 결과 형질전환체에서 acetyl esterase 활성을 확인했으며, 활성도는 숙주균주에 비해 약 4-5배의 높은 OD unit로 나타내는 것을 확인할 수 있었다.

본 연구에서는 $20^{\circ}C$에서 음이온 계면활성제인 AOT와 isooctane으로부터 형성된 reverse micelle을 이용하여 수용액상에 녹아있는 BSA인 모델 단백질을 분리하는 실험을 행하였다. 염의 농도가 0.1M에서 최고 가용화 효율은 KCl, NaCl, $MgCl_2$ 그리고 $CaCl_2$의 경우 각각 pH=5, 5.5, 7, 5 근처임을 알 수 있었다. 또한 매우 낮은 염 농도에서는 가용화는 일어나지 못하고 계면이 혼탁한 현상을 보였다. 특히 $CaCl_2$의 경우에 염 농도에 대하여 최고의 가용화 효율을 보였다.

In hybridoma cell culture, $NH_4{^+}$ is the most important toxic byproduct so far identified. It has been postulated that $NH_4{^+}$, which is similar to $K^+$ in size, is taken up non-specifically by the cells through a potassium transport system, and that the addition of $K^+$ to the culture medium may have a detoxifying effect of $NH_4{^+}$. Thus, in this article the effects of high $K^+$ concentrations in the range of 10 mM to 60mM on hybridoma cell growth and metabolism were investigated. No significant differences in growth were found for $K^+$ concentrations up to 40 mM, but cell death in the death phase was slightly delayed in the cultures with $K^+$ addition. At 60mM, growth was initially poor but the cells could be adapted after approximately 13 passages. With similar growth levels for high $K^+$ concentrations having been Identified in batch cultivations using basal medium, we are currently investigating how such high levels of $K^+$ will affect cell growth in fortified batch cultures where the accumulation of $NH_4{^+}$ is more problematical.

Studies on extraction of red pigment was performed to provide the basic information for the utilization of red pigment as s new source of natural food colorant. A bacterium isolated from marine resources were carried out the test for excretion of red pigment. One strain of a marine bacterium, KSR-97 showed a high production of red pigment on the medium of colloidal chitin, peptone-yeast extract with minerals. In physicochemical and sensory properties in aqueous solution of red pigment was investigated at various condition of pH, temperature, concentration of ethanol and stability of storage. Potent antioxidative of red pigment was separated by thin layer chromatograpy, silica gel chromatography and reverse high performance liquid chromatography using ODS hypersil column.

A neural network based controller (NN controller) was studied for the control of ammonia concentrations in biological processes. An ammonia FIA has been employed to on-line monitor the concentrations of ammonia in a bioreactor. The optimal neural network structure was investigated by computer simulation and found to be a 3(inputlayer)-2(hidden layer)-1(output layer). The NN controller had advantage over the PID controller, even though the former is more time consuming. The 3-2-1 NN controller has been used to control the ammonia concentrations in a simulated bioprocess and also in a real cultivation process of yeast, and its performance were investigated.

An algorithm for modeling of yeast fermentation process using genetic-fuzzy algorithm is presented in this work. The algorithm involves developing the fuzzy modeling of the process and model update capability against the system change. The membership functions of state variables and specific rates and the decision table were generated using genetic algorithm. This algorithm could replace the complex mathematical model to simple fuzzy model and cope with the change of process characteristics well.

The cooling rate of a bioreactor was measured to estimate the heat generation by microbial cultivation production. The estimated heat production was calculated from the varying temperature of cooling water. It was used for monitoring growth and specific metabolic events for microbial cultivations. Metabolic heat measured was also adopted for a control parameter for fed-batch cultivation.

Disinfection and molecular weight changes by electron beam irradiation has been examined. Before radiation, the viscosity of oven-dried HA decreased about 30% but the viscosity of vacuum-dried HA did not decrease. The number of viable cell depends on the relative humidity(RH). The cell number decreased about 30% in RH 0% and about 10% in RH 100%. After radiation, the decrease in the molecular weight of the hyaluronic acid was larger in the RH=0% than in the RH=100% and was larger in the vacuum-dried HA than in the oven-dried HA. The extent of disinfection were similar for all form.

광합성 홍색세균의 light harvesting complex II (LHC II)발현 유전자가 제거된 돌연변이종을 halogen 램프 하에서 거리에 따라 광도를 달리하며 배양하여 wild type과 생산성을 비교한 결과 낮은 광도(34 ${\mu}E/m^2/s$)에서는 wild type가 높은 광도 (376, 118 ${\mu}E/m^2/s$)에서는 mutant가 높은 세포농도를 나타내었다. 특히 118 ${\mu}E/m^2/s$에서 LHC $II^-$ mutant가 56% 높은 세포생산량을 보였다. 이는 세포내 pigment양의 감소로 mutual shading effect가 감소하였기 때문으로 판단되었다.

Production of eight avermectin components was improved in Streptomyces avermitilis wild type strain (ATCC31267) and high producing mutant strain (ATCC31780) when transformed with a foreign regulatory gene, afsR2 of Streptomyces lividans. Wild type and the high producing strain of S. avermitilis transformed with multiple copies of afsR2 improved total avermectin productions by 2.3 fold and 1.5 fold, respectively. In both of wild type and the high producing transformants carrying afsR2, glycerol was proved to be the best carbon source for the stimulation of avermectin production.

Optimization of submerged culture conditions for the production of exo-biopolymer from Paecilomyces japonica was studied. Maltose, yeast extract and potassium phosphate were the most suitable sources of carbon, nitrogen, and inorganic salt, respectively, for both production of the exo-biopolymer and mycelial growth. The optimal culture conditions in flask culture were pH 5.0, $25^{\circ}C$ and 150 rpm in a meidum containing of 30 g maltose, 6 g yeast extract, 2 g polypeptone, 0.5 g $K_2HPO_4$, 0.2 g $KH_2PO_4$, 0.2 g $MnSo_4\;{\cdot}\;5H_2O$, 0.2 g $MgSO_4\;{\cdot}\;7H_2O$ in 1-L distilled water. Exo-biopolymer production and mycelial growth in the suggested medium were significantly increased in a 2.5-L jar fermentor, where the maximum biopolymer concentration was 8 g/1.

Bifidobacteria의 배양효율을 증대하기 위하여 완충제로써 $CaCO_3$를 Ca-alginate에 고정화한 비드를 제조하여 사용하였다. B. longum KCTC 3218과 한국인 분변에서 분리한 B. longum HLC 3742 균주를 $CaCO_3$ 비드, NaOH, $Na_2CO_3$, $NH_4OH$ 등을 완충제로 각각 사용한 2.5-liter 발효기에서 각각 배양하여 균 증식과 저장에 따른 생존력을 조사한 결과 $CaCO_3$ 비드를 완충제로 사용한 경우가 다른 완충제를 사용한 경우보다 균 증식과 저장 안정성에서 효과적임을 알 수 있었다. 따라서 완충제로써 $CaCO_3$ 비드는 bifidobacteria의 고농도 배양과 생존력 증대에 유용할 것으로 사료되었다.

P. oleovorans의 유가식 배양에서 탄소원으로 octanoic acid, 질소원으로 $NH_4NO_3$를 이용한 혼합기질을 배양액의 pH 변화에 따라 공급하는 pH-stat 기질공급전략을 개발하였다. 공급기질의 탄소원/질소원 비 (C/N 비)를 변화시킴으로써 최종 균체농도, PHA 농도, PHA 함량 등을 변화시킬 수 있었으며, 최대 균체농도는 C/N 비가 10 (g octanoic acid/g $NH_4NO_3$)일 때 65 g/L, 최대 PHA 농도는 C/N 비가 20일 때 41 g/L, 최대 PHA 함량은 C/N 비가 20일 때 75%였으며 최대 PHA 생산성은 C/N 비가 10일 때 1.03 g/L/h였다.

Marine Microorganism strain 96CJ10356 produced extracellular polysaccharide (EPS-R) accompanied with cell growth. To improve the production of EPS-R, the effect of aeration rate was tested in a 5-liter jar fermentor with STN medium. The production of EPS-R was increased with aeration rate and after 72 hour cultivation, 12.20 g/l of EPS-R was obtained with an aeration rate of 1.5 vvm and the apparent viscosity was measured to be about 1000 cp with culture broth.

A. oryzae에 있어서 protoplast를 이용한 형질전환이 아닌 세포벽이 부분적으로 분해된 cell을 이용하여 electroporation으로 형질전환시켰고, novozyme234, hemicellulase와 celluclast를 사용하여 형질전환 효율이 어떻게 다른지를 비교 분석 하였다. Hemicellulase를 $^{\sim}10^8$ cell에 처리하여 A. oryzae에서 83 transformants/10ug of DNA를 얻었고, novozyme234와 celluclast를 사용하였을 때는 4.3 transformants/10ug of DNA를 얻었다.

Physiological changes of Escherichia coli during the fed-batch fermentation process were characterized in this study. Overall cellular protein samples prepared at the different stage of fermentation were separated by 2-dimensional gel electrophoresis (2-DE), and differently expressed 15 proteins, Phosphotransferase enzyme I, GroEL, Trigger factor, ${\beta}$ subunit of ATP synthase, Transcriptional regulator KDGR, Phosphoglycerate mutase 1, Inorganic pyrophosphatase, Serine Hydroxymethyl-transferase, ${\alpha}$ subunit of RNA polymerase, Elongation factor Tu, Elongation factor Ts, Tyrosine-tRNA ligase, DnaK suppressor protein, Transcriptional elongation factor, 30S ribosomal protein S6 were identified using matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). When bacterial cells grow to high cell density, and IPTG-inducible heterologous protein is produced, expression level of overall cellular proteins was decreased. According to their functions in the cell, identified proteins were classified into three groups, proteins involved in transport process, small-molecule metabolism, and synthesis and modification of macromolecules.

Batch cultivations of Anaerobiospirillum succiniciproducens have been systematically studied for the economical production of succinic acid from wood hydrolysate with corn steep liquor(CSL) as a nitrogen source. CSL was found to be an alternative complex nitrogen source for A. succiniciproducens when glucose and wood hydrolysate were used as carbon sources. Compared with polypeptone and/or yeast extract, CSL had similar effects on fermentation performance such as succinic acid yield and a ratio of succinic acid to acetic acid in the fermentation of wood hydrolysate as well as glucose. This means that succinic acid can be produced more economically from wood hydrolysate and CSL than relatively expensive carbon and nitrogen sources. Besides its low cost, the alternative medium served as a green technology for succinic acid production because it gives a net-zero effect on global warming.

본 연구에서는 담자균류의 일종인 Agaricus blazei를 이용한 polysaccharide의 생성을 증가시키고자 glutamic acid의 첨가시기의 영향을 비교${\cdot}$검토하였다. 그결과 2 g/L의 glutamic acid를 배양 4일째인 대수증식기에 첨가하여 polysaccharide 생성을 유도한 경우 배양 7일째에 12.9 g/L의 균체량과 9.1 g/L의 polysaccharide를 생성하였다. 그러나 이런한결과는 배양 초기에 glutamic acid를 첨가했을경우와 비교하여 볼 때 균체량은 다소 증가 했으나 polysaccharide 생성량에 있어서는 거의 유사한 결과를 나타내는 것이다. 생물반응기를 이용한 회분배양에 있어서도 2 g/L의 glutamic acid를 대수증식기인 배양 4일째에 첨가한 경우 균체량은 13.5 g/L, polysaccharide 생성량은 9.9 g/L를 생산하여 배양결과 최대 생산량을 나타내었다.

Paecilomyces japonica의 균체 생육 및 다당체 생성에 미치는 기본배지 및 물리적인 조건에 대한 영향을 조사하였다. 기본배지로는 균체량과 다당체 생성이 가장 좋은 YMP medium로 결정하였고 배양온도는 $27^{\circ}C$, pH는 9, 접종량은 2%로 선택하였다.

호기성 균주인 Streptoverticillium mobaraense에 의해 생산되는 Transglutaminase의 고수율을 위한 실험을 하였다. 라틴방격법에 의해 실험을 설계하여 최적 배지를 확정하였고, 호기성 균주인만큼 공기공급을 위한 통기 및 교반이 중요하게 작용됨을 관찰하였다. 이를 위해 임펠러의 형태 및 크기에 관한 실험을 수행하였고, 향후 부피산소공기전달 속도를 vvm과 교반속도에 따라 측정하도록 할 것이다. 또한 미생물의 증식 및 효소생산성에 미치는 온도 및 pH의 조건에 대해 실험한 결과 온도보다는 산성 및 중성에서의 pH가 효소생산성에 높은 영향을 미치고 있다. 플라스크 배양에서 평균적으로 1.3 U/mL 활성의 효소생산이 가능했으며 동일조건의 발효조 배양시 0.7 U/mL로 생산성이 감소하였다.

Clostridium.aetobutylicum Bl8에 의한 부탄올 발효에서 농도, extra nutrient, pH대한 부탄올 생성량을 알아보았다. 실험결과 extra nutrient를 주입했을 때 부탄올의 농도가 주입하지 않았던 배양에서보다 약 17%정도 증가하는 결과를 보였고, pH를 control 했을 때에는 glucose 60g/L 회분 실험을 기준으로 부탄올 생산량이 약 41%정도가 증가하는 것으로 나타났다.

Extracellular polysaccharide was produced by Pseudomonas elodea ATCC 31461 under aerobic condition. Nitrogen sources in medium affected cell growth and production of exopolymer. Ammonium nitrate limitation was found to be essential for higher production of exopolymer. Conversion rate of exopolymer from glucose under ammonium nitrate limitation was about 5 times higher than with ammonium nitrate.

Effect of carbon sources including agro-industrial byproduct on cell growth and production of curdlan by Agrobacterium sp. ATCC 31749 was investigated. Maximal production of curdlan was obtained when the carbon source was sucrose. The conversion rate of curdlan from 2% (w/v) sucrose was 59%. Glucose, mannose and maltose were also found to be good carbon sources for production of curdlan. Production of curdlan increased up to 3% (w/v) glucose as the carbon source and then decrease as the concentration of glucose increased. The major components of agro-industrial byproduct (AIB) were glucose, maltose, and maltose, and maltotriose. Agrobacterium sp.ATCC 31749 utilized up to 25% (v/v) AIB and produced curdlan with 29.8g/1.

The effect of phosphate on the production of erythritol and gluconic acid during the batch fermentation of Candida magnoliae SR101 was investigated. In the flask culture experiments, the results showed that phosphate concentration affected the production of erythritol and gluconic acid in Candida magnoliae. In the tormentor experiments, the increase of phosphate concentration of medium up to 10 g/L increased the gluconic acid, while the maximum erythritol concentration was 121.7 g/L from 250 g/L glucose and 3 g/L $KH_2PO_4$

The production of pullulan by Aureobasidium pullulans HP-2001 was investigated under various ratios of glucose as carbon source and yeast extract as the nitrogen source, Highest conversion rate (productivity) of glucose to pullulan was 40.0% when concentrations of glucose and yeast extract were 5% and 0.15%, respectively. Maximal production of pullulan was 29.3g/1 when the concentration of glucose was 8%(w/v) and that of yeast extract was 40:1. On basis of the result that production of pullulan was found in a medium which concentration of glucose as carbon source was up to 20%(w/v), Aureobasidium pullulans HP-2001 seemed to overcome the catabolite repression. Conversion rate of pullulan from 20%(w/v) of glucose was 11.1%.

The purified biosurfactant $3.16g/{\ell}$ was obtained after cultivation for 104hr at $25^{\circ}C$ with an optimal agitation speed of 200rpm, an aeration rate of 2vvm in a $14{\ell}$ fermenter containing $5.5{\ell}$ of LB medium and 1%(w/v) olive oil as a carbon source. For the kinetic studies, the optimal substrate concentration was analyzed on different olive oil concentrations(0.1, 0.5, 1.0, 1.5, 2.0%(w/v)) and optimal culture conditions(MLBM, 200rpm, 2vvm at $25^{\circ}C$) in a $14{\ell}$ jar fermenter. The results obtained indicate that $K_s$=0.0086 $g/{\ell}$, $q_s$= 0.664 $g/g{\cdot}h$, $q_p$= $4.2{\times}10^{-3}$ $g/g{\cdot}h$, and ${\mu}_{max}$ was determined as $0.1449h^{-1}$.

이차대사산물의 생산에 영향을 미치는 인자 중 통기성의 조절을 통한 Monascus 적색색소의 생산을 증가 시켰다. 높은 산소공급조절은 균체량을 증가 시켰으며, 낮은 산소공급조절은 고농도의 적색색소를 생산하였다. 따라서, 산소 공급량의 조절을 통하여 적색색소 생합성 과정을 활성화 시킬 수 있다.

A study was carried out to select a nitrogen source and the optimize the C/N ratio for the maximum cell growth of Phaffia rhodozyma in fed-batch culture. The yeast extract was the best organic nitrogen source. In the batch culture experiments, the highest cell yield was obtained 0.575 g-cell/g-glucose from 10 g/L and 10 g/L yeast extract. In the fed-batch experiments, the maximum cell concentration was obtained 33.1 g/L from the C/N ratio of 2:1 while the astaxanthin concentration of cell was Increased by increasing the C/N ratio, of feed medium.

본 연구에서 비스판균에 비해 아포형성능이 우수한 변이주를 선별하고, 발효조 배양에서 활성아포 생산성의 증가를 확인하였다. 최종 선별된 변이주 KD21균주를 5 L jar fermenter에서 배양한 결과, 36시간만에 총균수는 $5.5{\times}10^9$ CFU/mL에 도달하고, 최대 활성아포수는 $2.7{\times}10^9$ CFU/mL에 도달하였다. 이는 비스판 친균주에 비해 활성아포의 생산성이 77%증가하였음 확인 할 수 있었다. 장질환 치료제로 판매되고 있는 비스판 제제의 활성아포 생산성이 우수한 변이주를 획득함으로서 친균주에 비해 가격경쟁력을 가질수 있을것으로 판단된다.

S. lincolnensis 배양시 나타나는 거품의 문제를 해결하기위해 UP-1과 OP-1배지에 소포제 종류를 PPG, PEG 및 CA-110을 첨가하여 실험하였다. OP-1배지를 이용한 플라스크 배양의 경우 PPG와 PEG 각각 402 mg/L, 394 mg/L로 높은 Lincomycin 생산농도를 나타내었으나, UP-1배지의 경우 대조구에 비해 Lincomycin 생산농도는 큰 차이가 없었다. 두 가지 배지에서 소포제 농도의 변화에 대한 Lincomycin생산을 통계처리한 결과는 실험한 농도 범위에서 큰 차이를 나타내지 않았다. 충분한 산소공급은 Lincomycin 생산에 필수적이고, rpm의 변화에 따른 실험 결과 300 rpm의 경우 가장 효과적이었다. 발효조에서 PPG 농도 5, 1, 0.5 g/L를 첨가하여 발효한 결과 상당한 차이를 나타내었다. Lincomycin 생산을 위한 소포제로서 PPG가 가장 좋았으며 농도는 0.5 g/L가 적절하였다.

A. pullulans ATCC 42023를 사용하여 고분자량의 pullulan을 대량생산하기 위한 연구로써, 용존산소량과 pH가 균체 성장 및 pullulan 생산에 미치는 영향에 대하여 연구하였다. pullulan의 분해 효소에 대한 저항성을 가지고 산업적으로 유용한 고분자량의 pullulan을 생산하기 위해서 pH를 6.5로 조절할 경우, 분자량 20만 이상의 high molecular weight의 productivity가 가장 높았으며 용존산소량의 증가로 pullulan productivity를 증가시킬 수 있었다.

A. pullulans ATCC 42023의 pullulan생산과 형태학적인 변화를 알아보기 위해서 삼각플라스크와 회분식 발효를 이용하여 실험하였다. 삼각플라스크 내에서 초기 pH 6.5는 $11.98g/{\ell}$의 pullulan을 생산하였고, 회분식 발효에서는 pH 2.5-2.7의 범위에서 pH를 조절하면서 실험한 결과, pH 4.5에서 pullulan생산 $13.31g/{\ell}$의 최대값을 나타내었다. 그러나 균체성장은 pH 6.5에서 가장 높았다.

Thraustochytrium aureum ATCC 34034를 인공 해수 배지에서 $24^{\circ}C$, 200rpm, 초기 pH 6으로 고정하여 빛을 가하면서 3일간 배양을 한 결과, 최대 건조 균체량은 1.34g/L이었으며, 최대 DHA생산량은 41.4mg/L이었다. 모든 실험에서 탄소원으로 glucose를 5g/L로 고정하였다. YM배지와 YPG배지에서는 Thraustochytrium aureum이 DHA를 생산하지 못하였으며, 이 균주가 해양 미생물이라는 점을 고려해보면, 성장과 DHA생산에 있어서 염분이 중요한 영향을 미친다고 생각된다.

In order to maximize the RNA accumulation and biomass production in Saccharomyces cerevisiae MTY62, a high-content RNA yeast strain, fed-bach cultures were performed with optimized industrial medium including molasses and corn steep liquor. Among the feeding modes examined, the constant feeding mode resulted in the cell concentration of 35.7 g-DCW/L and the RNA concentration of 5434 ${\mu}g-RNA/mL$, which were about 2-fold increased levels, compared to the results of bach culture. However, the RNA content (153 mg-RNA/g-DCW) in the fed-batch cultures was lower than that in the batch culture (171 mg-RNA/g-DCW).

Actinoplanes teichomyceticus ATCC 31121 was mutated with UV to obtain a superior mutant strain with increasing level of teicoplanin production. In this investigation lethal curve was obtained and the optimal condition to induce mutagenesis was determined and to isolate the desirable mutant strain. It was also confirmed that teicoplanin activities by agar diffusion method to be compared to the mother strain. One mutant strain, T991014-1 that had the highest teicoplanin productivity was finally selected for further investigation including fermentation pattern. The mutant was characterized by the various tests such as amylase activity, protease activity, antibiotic resistance, autotoxicity, and productivity.

This study was observed to the effect of the feeding Platycodon grandiflorum A. DC (3 years) extract on the bronchus diseases bacteria ( C. diphtheriae, S. aureus, Mycobacterium sp., F. nucleatum, S. pygogenes, K. pneumoniae and N. gonorrhoeae) and fungi(A. fumicatus). Platycodon grandiflorum A. DC was extracted ethanon, water, ethyl ether and petroleum ether. The extraction rate of Platycodon grandiflorum A. DC to the extract solution was identified 71.8%, 100%, 15.4% and 14.1%. Each extract solution was injected culture media into several concentrations and then investigated the bacteria cell growth during 32 hours. As a result antimicrobial activity was excellent an extract by ethyl ether and petroleum ether. Among several concentrations, bacteria cell growth inhibition was observed from 0.06% to 0.14%. The rate of antimicrobial activity was over 70%. The cell growth inhibition rate of each bacteria was appeared in order of ethyl ether > petroleum ether > water > ethanol.

레트로바이러스를 동결 건조과정에서 발생하는 물리적, 화학적인 스트레스로 인한 바이러스의 실활을 보완하기 위해 첨가하여 준 여러 가지 첨가물 중 trehalose가 가장 탁월한 protectant임을 알 수 있었고 최적 첨가농도는 300mM이었다. Trehalose의 역할을 보완하기 위하여 분자량이 큰 polymer들을 첨가 하였으나 trehalose만을 넣어 주었을 때 보다 poly(vinylpyrollidone)(PVP)의 경우 큰 영향은 볼수 없었고 dextran과 PEG를 첨가 하였을 경우엔 오히려 더욱 활성이 감소하였다.

When 3 recombinant Chinese hamster ovary (rCHO) cell lines, CHO/dhfr-B-22-4, $CS13-1.00^{\ast}$ and $CSl3-0.02^{\ast}$, were cultivated in hyperosmolar media resulting from NaCl addition, their specific foreign protein productivity increased with medium osmolality. Glycine betaine was found to have a strong osmoprotective effect on all 3 rCHO cell lines. Inclusion of 15 mM glycine betaine in hyperosmolar medim enabled rCHO cell lines to grow at 557-573 mOsm/kg where they could not grow in the absence of glycine betaine. However, effect of glycine betaine inclusion in hyperomolar medium on foreign protein production differed among rCHO cell lines. CHO/dhfr-B22-4 cells retained enhanced specific human thrombopoietin (hTPO) productivity in the presence of glycine betaine, and thereby, the maximum hTPO titer obtained at 573 mOsm/kg was increased by 72% over that obtained in the control culture with physiological osmolality (292 mOsm/kg). On the other hand, enhanced specific antibody productivity of $CSl3-1.00^{\ast}$ and $CSl3-0.02^{\ast}$ at elevated osmolality decreased significantly in the presence of glycine betaine at a cost of the recovery of cell growth. As a result, the maximum antibody titer at 557 mOsm/kg was similar to that obtained in the control culture with physiological osmolality. Taken together, efficacy of the simultanous use of hyperosmotic pressure and glycine betaine as a means to improve foreign protein production was variable among different rCHO cell lines.

유전자 재조합된 Nicotiana tabacum을 이용하여 동결 보존 후의 세포 생장을 관찰하였다. 형질 전환된 Nicotiana tabacum의 동결 보존에서 회복배지에 계면 활성제와 산소 전달 물질을 첨가한 경우 첨가하지 않은 경우에 비해 세포 생장을 증대 시켰고 산소 전달 물질 보다 계면활성제의 영향이 크다는 결과를 관찰하였다.

식물세포의 장기보존방법 연구를 위하여 참당귀(Angelica gigas Nakai) 현탁세포를 이용하여 동결보존을 수행하였다. 최적의 예비성장배지로는 0.7 M sucrose를 첨가하여 6일간 배양한 것이 가장 좋았으며, 동결보호제로는 1 M sucrose, 1 M glycerol, 1 M DMSO를 혼합하여 사용하였을 때 가장 좋은 결과를 나타내었다. 동결 시에는 처음 8시간 동안 deep freezer에서 천천히 냉동을 한 후 액체질소에 넣는 것이 가장 좋은 것으로 나타났다. 또한 여러 가지 삼투압 증진제를 사용한 실험을 수행한 결과 sucrose가 가장 좋은 결과를 나타내었는데 이것은 sucrose가 삼투압 증진제 역할뿐만 아니라 cell membrane을 보호하기 때문으로 사료된다.

본 연구에서는, gas 성분의 조성을 on-line으로 자동제어할 수 있는 시스템을 확립하고, 이를 이용하여 각 성분들이 식물세포에 미치는 영향에 대해 조사하였다. 실험결과, $CO_2$가 첨가되었을 때, 세포의 생장에는 큰 차이를 볼 수 없었으나, 2차 대사물 및 polysaccharide의 생산성을 증대시켰다. 특히, polysaccharide의 경우, $CO_2$가 참가되었을 때 1.8배의 생산성 증대를 나타내었다.

BTX를 분해하는 미생물균주를 개발하기 위해 하수처리장과 VOC배출업소의 하수처리장에서 4종류의 활성슬러지를 채취하여 500 mg/L의 BTX에 3개월간 적응하였다. Benzene, toluene의 분해에는 Y혼합균주, p-, m-, o-xylene의 분해에는 A혼합 균주를 선별하였으며 각각은 단일 및 복합성분의 BTX를 빠르게 분해하였다.

황화수소와 암모니아 함유악취의 동시처리를 위해 Thiobacillus sp.IW가 함유된 3상 생물막유동층반응기를 사용하였다. 본 연구에서 황화수소는 미생물에 의해 산으로 전환되었으며 이어서 유입된 암모니아와 반응하여 황산암모늄이 생성됨으로서 양성분은 제거되었다. 황화수소의 경우 45 mg/l h의 유입량에서도 거의 완전한 처리 효율을 보여주었으나, 암모니아의 경우 유입량이 10 mg/l h에서 증가함에 따라 처리효율은 점차 감소하였다.

본 BAF 반응기 연구에서의 C/N비의 증가는 산소의 친화도가 큰 호기성 종속영양 미생물에게 유리한 조건을 형성하여 질산화의 효율을 감소시켰으나 공기양의 증가로 인해 질산화 미생물의 활성도는 다시 증가하였다. 총 질소의 제거율은 동일한 C/N비에서 공기량을 증가하여 많은 양의 암모니아성 질소가 질산성 질소로 산화될 때 증가하였으며 이 결과는 무산소조에서 많은 다공성의 여재에 미생물이 두꺼운 생물막을 형성하여 생물막 안쪽으로 용존산소가 쉽게 전달되지 않아 탈질에 큰 영향을 미치지 않은 결과라고 볼 수 있다.

Enzymatic deinking of office-waste paper was studied using crude cellulase and papain-hydrolyzed cellulase from Trichoderma reesei Rut C-30 in small-scale and mid-scale. The results were compared with deinkings using commercial enzyme(Novozym 342) and conventional chemical methods. Maximum brightness and freeness were obtained at 3 units/g Oven Dry Paper(ODP) of CMCase activity using crude cellulase in mid-scale deinking experiments. The deinked pulp had higher physical strength and brightness, and lower freeness and yield than the pulp deinked in small scale. In small scale deinking, maximum brightness and freeness were obtained at 2 unit/g ODP. Deinking by papain-hydrolyzed cellulase showed similar results with one by Novozym 342. It was better in brightness and freeness, but showed lower physical strength and yield, than the conventional deinking by sodium hydroxide. The ratio of endo-1,4-glucanase and exo-1,4-glucanase components in papain hydrolyzed cellulase from T. reesei Rut C-30 was similar to that of commercial enzyme, Novozym 342, implicating a successful application as a deinking enzyme.

1.3-Propanediol as a bifunctional organic compound could potentially be used for many synthesis reactions, in particular as a monomer for polycondensations to produce polyesters, polyethers and polyurethanes. Wastewater containing high concentration of glycerol was used to produce 1,3-propanediol in lower production cost which inturn the quantity of wastewater to be treated. In this study, various attempts were made to increase 1,3-propanediol production under different conditions by Klebsiella pneumoniae ATCC 15380. 1,3-Propanedio conversion yield and by-product formation were influenced significantly by pH and temperature. The Optimal glycerol and nitrogen concentration for 1,3-propanediol production were found to be 25g/L and 1%(w/v), respectively. The production formation of 1,3-propanediol was optimal at pH 6.0 and temperature $35^{\circ}C$.

Three 'stress probe' plasmids were constructed and characterized which utilize a green fluorescent protein (CFP) as a non-invasive reporter to elucidate Escherichia coli cellular stress responses in quiescent or 'resting' cells. Facile detection of cellular stress levels was achieved by fusion of three heat shock stress protein promoter elements, those of the heat shock transcription factor ${\sigma}^{32}$, pretense subunit ClpB, and chaperone DnaK, to the reporter gene $gfp_{uv}$. When perturbed by chemical or physical stress (such as heat shock, nutrient (amino acid) limitation, addition of IPTG, acetic acid, ethanol, phenol, antifoam, and salt (osmotic shock), the E. coli cells produced GFPuv which was easily detected from within the cells as emitted green fluorescence. A temporal and amplitudinal mapping of these responses was performed, demonstrating regions where quantitative delineation of cell stress was afforded.

Biosorption process is an economic and potential process for metal sequestering from the water. The oxidized Undaria pinnatifida by nitric acid had high uptake capacity for heavy metals of 4 - 6 meq / g dry mass. For the application of oxidized Undaria pinnatifida, recovery of metal in plating wastewater was studied. The uptake capacity of the oxidized Undaria pinnatifida was high compared to the ion exchanger IR-120 plus. The treatment efficiency of chromium and copper in the wastewater was 85% In batch. Activated carbon was used to assist the recovery of water by removing organic matters of the wastewater.

To remove nitrogen compound from wastewater six kinds of bacillus were isolated from sludge. Each bacillus was identified as B. subtillis $I{cdot}II$, B. cereus $I{cdot}II$, B. anthracis, B. circulans. The test of effect of nutrient and cofector on the nitrogen removal showed that peptone, yeast extract, magnesium, iron, and calsium accerated the efficiency of nitrogen removal. In syringe test aerobic nitrification and denitrification was occured.

반응기의 performance를 결정하는 중요한 요소는 methane 생성균의 활성을 측정하는 것이다. Methanogen의 활성을 측정하는 방법으로 SMA가 유일하게 사용되고 있다. 따라서 본 연구에서는 methanogen의 활성을 측정하는 다른 대안으로 CLSM image의 정량 분석 방법을 확립하였다. CLSM의 결과는 약 5시간 안에 얻을 수 있을 뿐만 아니라, 분자 생물학적인 기술을 많이 요하지 않는다. 또한, sludge내부의 상태와 methanogen의 분포정도를 사진으로 볼 수 있는 장점이 있다. 따라서 SMA와 함께 CLSM image 정량 분석은 UASB반응기의 start-up 동안의 methanogen의 활성을 측정하기 위한 유용한 방법으로 제안할 수 있다.

In this work, an extractive membrane bioreactor containing coulture broth of Burkholderia cepacia G4 PR1 constitutively expressing the TCE-degrading enzyme, tolune-ortho-monooxygenase(TOM), was used for the degradation of TCE. The membrane bioreactor operates by seperating the TCE-containing waste gas from the aerated biomedium, by which the air-stripping of TCE without degradation was overcome that could occur in conventional aerobic biological treatments of TCE-contaminated waste gases. This was achieved by a silicone rubber membrane which was coiled around a perspex draft tube. TCE from the gas phase diffuses across the silicone rubber membrane into microbial culture broth that was continuously fed from a separate aerobic CSTR. Therefore, TCE degradation occured without the TCE being directly exposed to the aerating gas stream. Of the TCE supplied to the membrane bioreactor, 72.6% was biodegraded during the operation of this system. To construct a mathematical model for this system, parameters describing microbial growth kinetics on TCE were determined using a CSTR bioreactor. Else parameters used for numerical simulation were determined from either indepedent experiments or values reported in the literature. The model was compared with the experimental data, and there was a good agreement between the predicted and the measured TCE concentrations in the system. To achieve a higher treatment efficiency, various operating conditions were simulated as well.

본 연구에 사용한 Pseudomonas sp. BCNU 154는 Gram 음성 toluene 내성세균으로 형태적${\cdot}$생리적 특성을 비교하여 보았을 때 Pseudomonas 속임이 확인되었다. 폭넓은 유기용매에 대한 내성한계를 가지고 있으며 Pseudomonas sp. BCNU 154 toluene 내성균주는 Modified Luria-Bertani(LBM) 배지에서 잘 자랐으며, $30^{\circ}C$에서 최적의 생육상태를 나타내었다. 최적 pH는 6에서 7로 나타났으며 1%의 toluene을 첨가하였을 경우에는 pH 2에서 8에 이르는 넓은 pH대에서 잘 자랐으며, cumene은 pH 6에서 10까지의 범위에서 좋은 성장을 보였다. Pseudomonas sp. BCNU 154 균주의 toluene, ethylbenzene, ${\rho}$ -xylene, cumene분해는 $1g/{\ell}$의 농도로 첨가시 toluene의 경우 97%이상이 분해되었고, ${\rho}$은 92%, ethylbenzene은 80%이며, cumene은 90% 이상이 분해되는 것으로 나타났다. 따라서 각 유기용매에 대해 대체로 12시간 내에 90%이상의 분해가 이루어졌으며, ethylbenzene의 경우에는 성장곡선과 마찬가지로 일정시간이 지난 후 성장과 분해가 둔화되다가 다시 성장과 분해가 일어나는 현상을 보였다.

The optimal conditions and properties for the immobilization of marine bacterium Pseudomonas aeruginosa BYK-2 have been determined. For the high productioon of biosurfactant, Na-alginate, PVA, modified PVA were used as a carrier. The optimal emulsifying activity on immobilized Pseudomonas aeruginosa BYK-2 showed 1036Unit (about 2.2g/L biosurfactant) in Basal salt medium(B.S.M.) at $25^{\circ}C$, 100rpm. Ca-alginate was selected the optimal bead among PVA, modified PVA and Ca-alginate. The optimal cell load in alginate bead was 10 gCWW/100g carrier. As the results of incubation of immobilized 5g Ca-alginate bead (conditions; 3% alginate, bead diameter: 2.3mm, 10% cell load) in 50m1 production medium, The emulsifying activity of 1407Unit, about 3.0g/L biosurfactant was obtained from immobilized cell after cultivation of 92hr at $25^{\circ}C$, 100rpm.

본 연구에서는 염색폐수의 생물학적 처리를 위하여 염색폐수에서 선별된 균주와 현재 상용화된 담체를 이용하여 실험하였다. 그 결과 단일 균주에서는 평균 45%의 색도 제거율을 보였다. 2균주 조합을 사용했을 때는 색도 제거율이 86%까지 증가하였다. 담체는 충진율 15%일 때 최적을 보였다.

Polysaccharides are very effective adsorbents for heavy metals. In this study, the adsorption characteristics of various polysaccharides for heavy metal adsorption were investigated. Tested polysaccharides were homogeneous polysaccharides such as curdlan, chitin, starch, cellulose, Avicel, and Solka floc and heterogeneous polysaccharides such as zooglan, locust bean gum, ghatti gum, pectin, and xylan. Lead(II) adsoption characteristic on these polysaccharides followed Freundlich isotherm and the isotherm parameters were calculated. For adsorption of lead(ll), Avicel, starch, and zooglan were found to be good adsorbents.

재조합 발광미생물인 UV2와 YH9을 보존제로서 trehalose를 혼합한 현탁액을 바로 동결건조한 것과 이 현탁액을 $30^{\circ}C$ incubator에서 15분 동안 진탕배양한 후 동결건조한 것과 trehalose가 첨가된 배지에 약 $6{\sim}7$시간 정도 배양한 배양액을 동결건조한 결과, trehalose를 첨가한 후 15분 동안 진탕배양했을 경우가 UV2는 0.08M의 이상에서 약 45.0%, 50.0%, YH9은 0.16M에서 48.7%, 54.1%로 회복율을 가장 높이는 최적의 조건으로 조사되었다.

Styrene을 제거하기 위한 biofilter가 연구되었다. 운전 초기에 활성슬러지를 접종함으로써 start-up 기간을 24시간으로 줄일 수 있었다. 운전중 질소원 고갈 현상이 발생하였고 첨가된 ammonium sulfate양에 따라 제거된 styrene의 양을 정량적으로 구하여 이것을 biofilter의 장기운전에 이용할 수 있었다. Styrene의 maximum elimination $capacity(EC_{max})$와 critical elimination $capacity(EC_{cr})$는 각각 4.8kg $C\;/m^3{cdot}day$, 1.248kg $C\;/m^3{cdot}day$이었으며 styrene 농도 400ppmv까지를 분해하는데 EBRT 1min으로 제거율 95% 이상을 달성할 수 있었다.

A device to measure the temperature difference between the supernatant and the sediment blanket in the course of SV30 measurement in processing of activated sludge process. The temperature elevation in the sludge sediment represent the metabolic heat production by the microorganisms and can be an indicator for the capability of waste treatment. The utilities of the device for the analysis of activated sludge process were demonstrated in this report.

Batch culture system and continuous culture system were used to investigate the removal of disperse dye using several white rot fungi. White rot fungi used in the study were Coriolus hirsutus IFO 4917, Lenzites betulina IFO 6266, Coriolus versicolor IFO 30340 and Phanerochaete chrysosporium IFO 31249. The results of the batch culture experiment showed that white rot fungi used in this study had excellent dye removal abilities. Phanerochaete chrysosporium IFO 31249 was especially effective on the removal of disperse dyes. And continuous treatment of disperse red-60 was studied under bioreactor with vertical matrix using Phanerochaete chrysosporium IFO 31249. The removal efficiency of disperse red-60 were more than 95% in 0.20 ${\sim} 1.50 $hr^{-1}$ dilution rate and 90% in $1.83h^{-1}$ dilution rate.

본 연구는 T시에서 다량으로 배출되고 있는 종합염색폐수의 생물학적인 처리방법을 기존의 활성슬러지조에 비해 별도의 산기장치가 없는 상태에서도 산소전달효율이 뛰어나며, 기포의 표면적이 작아 용존 되어지는 산소의 양이 많고 또한, 저동력비을 갖는 JLR를 사용하여 새로운 생물학적 처리 공정을 확립하고자 하였다. 이를 이용하여 BOD의 제거효율이 짧은 8시간의 체류시간에서도 99%에 이르는 뛰어난 유기오염원의 제거효율을 얻었다. 산업의 발달로 의식수준이 점차 높아져감에 따라 생활환경이 개선되어 다량의 난분해성 물질이 주요원인인 색도가 수질의 큰 문제점으로 대두되었는데, 이를 제거하는 방법에 있어서도 기존의 1차 응집공정의 과량의 응집제 사용으로 인한 대량의 슬러지생산과 처리비용 등에 관한 문제점을 효율이 뛰어난 활성탄 담체를 포함한 JLR로서 1차 처리함으로써 응집제의 양의 감소와 87%에 달하는 색도의 제거효율을 얻을 수 있었다. 이를 다시 2차 응집하면 약 2,300의 유입수를 150으로 94%정도의 제거효율을 얻어 수질의 향상시킬 수 있었다.

TBR에 공급되는 성장기질인 phenol의 농도에 따른 TCE 분해 효율 변화를 살펴보았다. Phenol 공급이 없는 경우에서의 TCE 제거효율은 평균 42.8%로 유지된 반면, phenol을 0.94 ppm 농도로 공급해줌으로써 TCE 분해 효율을 16% 정도 증가시킬 수 있었다. 따라서 TBR에 알맞은 양의 성장기질을 공급하면, TCE 분해 과정에서 불활성화되는 효소 및 세포의 재활성화를 통해 TCE 분해 효율이 증가시킬 수 있었다.

기존의 BACC process의 가장 큰 단점은 탈질이 잘 이루어지지 않는다는 점인데 이것을 보완한 modified BACC process의 경우 실제오수를 사용하여 외부 반송비 따른 질소 및 인의 제거율을 살펴보면 외부 반송비가 200%일 때 $CODC_{Cr}$의 제거율은 평균 $96.3{\sim}95.7%$ 기존의 BACC process와 비슷하나 T-N 제거율은 $88.3{\sim}95.7%$로 월등히 우수한 결과를 보여주고 있다. 충진율 실험에서는 Table 2에서 보는 바와 같이 큰 차이는 없었다.

A model was developed to describe the microbial decontamination of diesel contaminated soil in a soil column. The biodegradation rate of diesel in nature depends on temperature and the pH of soil, availability of nutrients, oxygen and water. The soil moisture content is one of the essential factors because it characterizes the availability not only of water to microorganisms but also of oxygen and nutrient dissolved in soil. In this work, the rate of biodegradation was modeled by coupling Michaelis-Menten kinetics for the aqueous-phase solute with adsoption-desoption equation for diesel sorption and desorption from soil.

The protozoan parasite Giardia lamblia has been implicated as the causative agents of many outbreaks of waterborne intestinal illness. Accurate evaluation of Giardia lamblia removal in water treatment process requires a reliable method for measuring the concentrations of these pathogens in water. The relative recovery of Giardia cysts was assessed for seeded samples of distilled water. Cysts preparation was done by encystment in vitro. Membrane filtration was evaulated with cellulose acetate, polycarbonate, polypropylene, polyethersulfone, nylon membranes. Elution conditions were varied to improve cyst recovery.

The effects of the presence of commercial non-ionic surfactants on the cell growth and diesel degradation by Pseudomonas sp. OSD were studied. Most surfactants inhibited the diesel biodegradation at high concentration(1000mg/1). However, some surfactants showed no inhibition at lower concentrations. Tween 20, Brij 58, Brij 78 were not inhibitory to the diesel biodegradation even at high concentration. These chosen surfactants has relatively high HLB values. There exists complicated relationship for diesel bioremediation between cell hydrophobicity, surfactant HLB, contaminants, an soil.

Recombinant E. coli JM109(pZH3-5/pMT) harboring manganese transport gene(mntA) and metal sequestering protein, metallothionein(MT), was cultivated to accumulate cadmium in aqueous phase. Bioaccumulation followed Michaelis-Menten type kinetics. Equilibrium isotherm showed Langmuir type isotherm. The optimum pH for $Cd^{2+}$ uptake was 7-7.5.

The disinfection effects of Trichloro isocyanuric acid (TICA) and Calcium hypochlorite (Cal-hypo) against E. coli' in aqueous suspension were compared at various concentrations of disinfectants as well as exposure times. When E, coil($^{\sim}10^7$ CFU/ml) were exposed by TICA and Cal-hypo (12 ppm each), 90% of the initial cells were reduced in 4 sec and 390 sec, respectively. Although Cal-hypo lost its disinfection capability in about 1 hr under the sun light, TICA maintained its effect up to 6 hrs. This comparative studies clearly demonstrate that TICA is more effective than Cal-hypo in terms of sterilizing E. coli as well as maintaining the disinfection effects.

Streptomyces setonii(ATCC 39116) is a thermophilic gram-positive soil bacteria which undergoes an ortho-cleavage pathway in the presence of phenol or benzoate as a sole carbon and energy source. The specific activities of catechol-1,2-dioxygenase in S. setonii, a key enzyme in ortho-cleavage pathway, were induced by various aromatic compounds such as benzoate, phenol, m-hy-benzoate, p-hy-benzoate, catechol, o-cresol, m-cresol, p-cresol, benzene, toluene, ethyl-benzene, 2-chloro-phenol, and 4-chloro-phenol, among which the phenol showed the highest inducibility in the presence of 0.01% glucose. More than 0.1% glucose dramatically reduced the specific activities of catechol-1,2-dioxygenase induced by most of the single aromatic compounds tested.

Ozone has the advantages for the strong oxidant agents. Ozone is widely used as a disinfectant and deodorant in water treatment and biosafety cabinets. We attempted deodorization and disinfection in the piggery using ozone treatment. It was found that ozone affects deodorization of ammonia and decrease of microbial organism in the air. Concentration of ammonia decreased 50 percents and microbial organisms in the air were decreased below 10 percents. It will be useful for the application of the ozone to environmental treatment in the piggery.

Amphiphilic polyurethane(APU) particle is a polymeric surfactant, and could increase the solubility of 2-methylnaphthalene significantly. 2-Methylnaphthalene was recovered by the precipitation of APU particles and was degraded by Acinetobacter sp. K2-2. APU particle was recovered and reused after treatment of triethylamine.

In this study, we investigated the theoretical and experimental study for separation of solid-liquid suspensions of water and fine particles using acoustic standing wave. When the acoutic force was not applied, the separation efficiency was decreased as flow rate was increased. When it was applied, the separation efficiency was maintained over 95%.

일반적으로 시중에서 유통되는 콩나물은 시루 채 출하되거나 200-300g 정도의 소포장으로 팔려지게 되며 상품으로써의 생명력도 3-4일 정도로 짧다. 본 실험은 콩나물의 저장 및 유통기간 연장방법의 일환으로 이온수 처리에 따른 저장기간 중의 변화를 조사하였다. $4^{\circ}C$와 $10^{\circ}C$에서의 이온수 처리 콩나물은 1/2 ${\sim}$ 1/3배 정도의 낮은 부패균 밀도를 나타내었으며, 실온과 가까운 $25^{\circ}C$에서도 1/2배의 낮은 부패균 밀도를 나타냄으로써 저온저장에서의 저장성 향상뿐만 아니라 콩나물의 실온유통에서도 하루정도의 저장기간 연장효과를 가져올 것으로 평가된다.

감귤의 저온저장 중 부패방지를 위한 천연항균제 개발의 일환으로 감귤의 저온저장 중 과피로부터 분리한 곰팡이를 MEA와 PDA 배지에서 배양하면서 분생자병과 분생자의 형태적 특성을 조사한 결과 플라스크형의 phialide, 분생자병의 branching type이 simple type이며, conidial head가 columnar shape 등의 특징을 갖는 Penicillium sp. CF-301로 동정되었다. 또한 국내 자생식물을 채집하여 유기용매로 추출한 후 향균활성을 조사한 결과 측백나무 열매와 주목나무를 제외한 8종의 식물 추출액이 감귤부패균에 대하여 향균활성을 보였으며, 특히 측백나무 즙액은 8mm의 투명환을 보여 가장 높은 항균활성을 나타내었다.

The pretreatment of used newspaper for the enzymatic digestion preprocess was performed on a percolation reactor and a batch reactor. The test condition of percolation process was $170^{circ}C$, 60min, 1 mL/min, and 400psi, that of batch was $40^{circ}C$, 3hr. and latm Reaction solutions used in pretreatment process were aqueous ammonia, sulfuric acid, water, and hydrogen-peroxide as an oxidizing agent. As a result, the effect of pretreatment was similar to batch and percolation process, but the yield of enzymatic hydrolysis was higher in batch than percolation. This batch pretreatment enhanced enzymatic hydrolysis rate and increased glucose yield from about 15 to 20%. The inhibition factors influenced the rate of enzymatic hydrolysis was investigated, and the ink contented newspaper was the major factor.

부분 정제한 VHb-DAO와 CPC를 30분 이상 반응시키면 CPC가 GL-7ACA로 80%이상 변환되었다. VHb-DAO와 D-form의 아미노산이 반응한 결과 $K_m$은 D-Methionine, D-Valine이 낮으며, $V_{max}$는 D-${\alpha}$-aminophenylacetic acid, D-Leucine, D-Phenyalanine가 높은 수치를 나타내었다.

Indole은 미생물에 강한 독성을 가지고 있으므로 indole의 생물학적 전환이 어려우므로 단지 소수의 미생물들이 Indole이나 그것의 연관물질들로부터 Indigo와 Indirubin을 생산하는데 응용되어 왔다. 본 연구실에서 분리된 Pseudomonas savastanoi BCNU 106은 toluene에 강한 내성을 가졌으며 P. savastanoi BCNU 106의 Indole 내성 정도는 p-xylene이나 toluene이 배지 부피의 20% 로 첨가되었을 때 160 mg/ml로 매우 높은 내성 정도를 나타내었다. Indole (100 mg/ml) 과 p-xylene (0.2 ml/ml)을 포함한 two-phase culture system에서 자란 P. savastanoi BCNU 106은 indole로부터 푸른색 indigo나 보라색 indirubin의 미생물학적 전환을 보여주었다. indigo와 indirubin의 생산은 P. savastanoi BCNU 106가 적절한 농도의 유기용매가 중층된 two-phase system에서 자랐을 경우에만 형성되었다. 그러므로 본 연구는, 분리된 indole 내성 세균이 유기용매하에서 indirubin과 같은 산업적 가치를 가진 유용물질을 생산하는데 응용될 수 있을 것으로 사료된다.

P. savastanoi BCNU 106 균주에의해 cholesterol 이 전환됨을 확인하였고 toluene에서보다는 p-xylene 에서 전환율이 높음을 관찰할 수 있었다. 또 균주 배양후 cholesterol이 녹아있는 유기용매를 첨가하였을 때가 전환율이 높음을 확인 할 수 있었다

The INU2 gene encoding an endoinulinase of Aspergillus ficuum was expressed by the Kluyveromyces marxianus INU1 promoter in a SUC2-deleted Saccharomyces cerevisiae to produce the endoinulinase free of an exoinulinase and an extracellular invertase in the culture medium. When inulin was included in the medium, a recombinant yeast strain produced the sufficient amount of the enzyme to make a halo around its colony. An expression of endoinulinase was dependent on the culture temperature and shaking. The highest expression of endoinulinase was observed at $30^{\circ}C$, and 150rpm.

Thiobacillus thioparus TK-m에서 메칠머캅탄(MM) 가스를 기질로 메칠머캅탄 산화효소(MMO)를 유도할 수 있었으며 DEAE-Sephacel과 Superose 12 컬럼 크로마토그래피를 이용하여 정제하였다. 이때 얻어진 각 효소의 비활성은 각각 374과 1240.8 units/mg-protein이었으며 효소의 최적 온도는 $55\;^{circ}C$였다. 상기 정제된 효소액은 SDS-PAGE상에서 66.1 kDa의 분자량을 가지며 20 mM $(NH_4)_2SO_4$ 혹은 200 mM NaCl의 첨가에 대해 최대 활성을 보였다. 글리세린, 에탄올, 아세톤의 첨가에 대해 효소활성은 부분적으로 불활성화되었으며 메탄올에 대해서는 저해받지 않았다. Purpald 발색법을 이용한 흡광도의 변화는 구강내 존재하는 MM 가스로부터 생성될 수 있는 수 십 nmol의 범위의 포름알데하이드에 대해 눈으로 인지가능한 발색시스템에 사용할 수 있음을 확인하였다.

발효액으로부터 유산을 정제하기 위해서 나노여과를 이용할 때, hardness의 제거효율은 발효액내 이온성분의 분포와 밀접한 관련이 있다. 특히 음전하를 띠고 있는 막의 경우, 용액내 이가 음이온에 의해서 모든 이온들이 영향을 받으며 일가 음이온의 반발력은 거의 없는 것으로 관찰되었다. 따라서 발효액내 일가 음이온의 양을 줄이거나 더 높은 전하밀도를 가진 막을 사용한다면 나노여과를 이용하여 발효액내 hardness를 효과적으로 제거할 수 있으리라 생각된다.

The extraction efficiencies of panaxynol and panaxydol according to extraction temperature were the highest at $80^{cdot}C$ among 65, 80, $95^{cdot}C$ with soxhlet and increased with shaking method when temperature increased from $25^{cdot}C$ to $45^{cdot}C$. The amounts of panaxynol and panaxydol were determined by gas chromatography. In time dependence of extracted amounts of panaxynol and panaxydol using shaking method, the efficiencies of panaxynol and panaxydol were increased during 12 hour. The effect of water swelling on panaxynol and panaxydol extraction efficiency using soxhlet and shaking methods, the efficiencies of panaxynol and panaxydol were decreased as swelling time was increased.

유류오염토양에서 분리한 유류분해균주 중 계면장력 저하능이 우수한 균주를 분리하여 동정한 결과 Pseudomonas aeruginosa D2D2로 판명되었다. 탄소원으로 glucose와 olive oil을 각 1.5%씩 첨가하였을 때 표면장력이 27.9 dyne/cm로 표면장력 저하능력이 가장 우수한 것으로 나타냈다. 생산된 biosurfactant로 ethly acetate 추출하여 crude biosurfactant를 얻었으며, TLC 분석한 결과 glycolipid계열의 biosurfactant임을 확인하였다.

The purification of a thermostable esterase expressed in Escherichia coli was investigated using thermoprecipitation of unclarified cell homogenates followed by after applying the heat-treated lysate to phenyl-sepharose column, and elution with detergent. Heat treatment at $70^{cdot}C$ was capable of removing to E. coli proteins. Specially, the thermoprecipitation with 15% polyethylene glycol 8000 can remove host proteins and nucleic acids efficiently. Various detergents were used to recover the esterase, which was strongly bound to phenyl-sepharose resin. Triton X-100, non-ionic detergent, was found to be the most efficient of all tested detergents.

In order to determine the position and the content of fatty acids attached to glycerides and the migration degree of fatty acids in the migration reaction, hydrolysis of fish oil was carried out with lipolase-100T derived from Aspergillus oryzae. The content of fatty acids in the glyceride mixture was analyzed and compared with that of fish oil. The amounts of fatty acid in 2-position and the migration degree of the fatty acid in 2,3-DG (diglyceride) and 2-MG (monoglyceride) were calculated. The results showed that approximately 95% (w/w) of DHA (docosahexaenoic acid) and 65% of EPA (eicosapentaenoic acid) was attached to the 2-position of glycerides in the fish oil. Approximately 87% (w/w) of DHA and 75% of EPA remained in 2,3-DG and 88% of DHA and 65% of EPA in 2-MG were not involved in the migration reaction.

An antifungal antibiotic was produced from Bacillus sp. YJ-63, and antibiotic was purified to a homogeneity by butanol extraction, Silica-gel column chromatography, Sephadex LH-20 gel filtration column chromatography and HPLC. The purified antibiotic showed one spot on Thin Layer Chromatography, which was negative to ninhydrin and positive to iodine. This finding indicated that the antibiotic was a cyclopeptide structure. UV absorption spectrum of the antibiotic in methanol was maximal at 277 nm, which corresponded to the spectrum for all of the Iturin antibiotic. The antibiotic was stable up to $121^{cdot}C$ and in the range of pH $3.O{\sim}l2.0$ with a stable of ${\alpha}-amino$ acid and ${beta}-amino$ acid, and showed high activity for fungi and yeasts,

일정전류와 일정전압조건 모두에서 완충액 농도가 증가할수록 시료들의 이동시간이 증가하였으며 전압이 높을수록 시료들의 이동시간이 단축됨을 알 수 있었다. ALA, PBG, glycine, LA의 최적 분석조건은 borate 완충액 30mM, 20kV, $76{\mu}A$에서 가장 잘 분리 되었다. 본 연구를 통하여 ALA, PBG, glycine, LA를 동시에 분리할 수 있을 뿐만 아니라 ALA 2mM, PBG 0.25mM 범위내에서 선형적인 보정곡선을 보이므로 정량분석이 가능하였다. CE를 이용한 ALA와 PBG의 동시 정량분석으로 인체, 동식물 및 광합성 미생물의 대사과정에 수반되는 효소인 ALA synthetase, ALA dehydratase의 활성 연구, 광합성 홍색세균에 의한 ALA생합성, 의학, 생화학, 효소공학 등에 응용할 수 있을 것으로 기대된다.

초임계 이산화탄소에 의한 동결 건조된 췌장내의 지질 추출 속도는 추출도, 추출압력, 초임계 이산화탄소의 유량, 원료의 입자크기에 의존하였고, 추출평형은 충진된 원료의 양에 대한 초임 계 이산화탄소의 유량에 비례하여 추출시간의 조절이 가능하였다. 초임계 처리된 원료와 수입 판크레아틴의 효소 역가 측정결과 수입원료에 비해 약 1.8배 이상의 높은 역가를 보였으며, 추출된 콜레스테롤 함량을 분석한 결과 용매추출을 한 경우는 약 2.1mg/g, 초임계 처리하였을 때에는 약 $2.2{\sim}3.1mg/g$정도 함유되어 있었다.

A novel cell surface display system was developed by employing the Escherichia coli outer membrane protein C (OmpC) as an anchoring motif. Poly-histidine (poly-His) peptides of 19, 32, 45, 84, and 162 amino acids (aa) could be successfully displayed by inserting them into the seventh exposed loop(L7) of OmpC. Recombinant cells displaying poly-His of 19, 32, 45, and 84 aa could absorb 18.9, 23.9, 26.1, and 32.0 ${\mu}mol$ of $Cd^{2+}$ per gram cell dry weight, respectively and therefore, would be useful as the biosorbents of heavy metals.

중국산 야생키위로 total RNA를 추출하고 RT-PCR를 실시해서 증폭된 1.2kb fragment를 얻어 pGEM-T Easy에 cloning한 후 염기서열과 아미노산 서열을 비교 분석한 결과 actinidin gene인 것으로 분석되었다.

The fumB gene encoding anaerobic fumarase of Escherichia coli XL1-Blue was introduced to solve the malic acid accumulation problem. When NZN111 harboring pTrcMLFu was cultured, 7 g/L of succinic acid was produced and malic acid was not accumulated.

Human leptin is a 16 kDa (146 amino acids) protein secreted from adipocytes and influences body weight homeostasis. In this report, active human leptin was successfully produced in the culture medium of Bacillus subtilis. After simple purification steps consisting of ammonium sulfate precipitation and anion-exchange column chromatography, 2.3 mg of leptin with a purity of greater than 95% was obtained from the 0.5 L culture with the recovery yield of 54.9%. The purified leptin showed the correct folding structure with one disulfide bond.

For the sustained release formulation of recombinant human growth hormone (rhGH), dissociable rhGH aggregates were microencapsulated within poly(D,L-lactic-co-glycolic acid) [PLGA] microparticles. rhGH aggregates with 2 - 3 m Particle diameter were first produced by adding a small volume of aqueous rhGH solution into a partially water miscible organic solvent phase(ethyl acetate) containing PLGA. These rhGH aggregates were then microencapsulated within PLGA polymer phase by extracting ethyl acetate into an aqueous phase pre-saturated with ethyl acetate. The resultant microparticles were 2 - 3 m in diameter similar to the size of rhGH aggregates, suggesting that PLGA polymer was coated around the protein aggregates. Release profiles of rhGH from these microparticles were greatly affected by changing the volume of the incubation medium. The release rhGH species consisted of mostly monomeric form with having a correct conformation. This study reveals that sustained rhGH release could be achieved by microencapsulating reversibly dissociable protein aggregates within biodegradable polymers.

Avidin-biotin의 강한 결합력을 이용하여 금속 표면 위에 리포좀과 같은 유기 분자막의 다층 형성 과정을 QCA의 공진주파수와 공진저항의 변화를 측정하므로서 실시간 모니터링의 가능성을 검토하고, 유기 분자막이 금속 표면 위에 적층 됨에 따라 형성되는 적층 막에 대한 정보를 수집함으로서 바이오센서 시스템으로 QCA를 적용 가능함의 기초 데이터를 제공하고자 한다.

The polymeric matrices made with poly(D,L-lactide-co-glycolide) were prepared using copolymer of poly(D,L-lactide) and poly(ethylene glycol) for application of drug delivery systems. Catalyst made use of stannous actoate. Particle size were differ greatly$(435.3{\pm}11.2{\sim}2284.1{\pm}188.5)$ that nanoparticle made use of according to solvent of various kinds. Polymer could a sharp distinction with copolymerized among LE-1, LE-2 and LE-3 of PLA and PEG of content that to examine $^1H-NMR$ of copolymer make refine and reprecipitation. Drug delivery effect at PLGA nanoparticle : PLA amount more then proved highly drug delivery amount that each LE-1, LE-2, LE-3, drug and solvent was 40mg, 20mg and 10mg. Drug delivery effect proved higher 20mg that change(10mg, 20mg, 40mg) at drug feeding amount with LE-2. The first a lot of drug proved delivery. LE-3 most lactide content proved much delivery since biodegradable on PLGA copolymer result from lactide. Also biodegradable rate was highest at LE-3 much of lactide content, because influence at biodegradable effect of lactide by inclusive of soft PEG.

Strain JV3 was isolated from commercial Swiss cheese products and identified as a bacteriocin producer, which has bactericidal activity against Leuconostoc mesenteroides KCCM 11324. Strain JW3 was identified tentatively as Lactococcus lactis by the API test. The activity of lacticin JW3, named tentatively as the bacteriocin produced by Lactococcus lactis JW3, was detected during the mid-log growth phase, and reached a maximum during the early stationary phase, and decreased after the late stationary phase. Its antimicrobial activity on sensitive indicator cells was completely disappeared by protease IV. The inhibitory activities of lacticin JW3 were detected during treatments of up to $121\'^{circ}C$ for 15 min. Lacticin JW3 was very stable over a pH range of 2.0 to 9.0 The apparent molecular mass of lacticin JW3 was estimated to be in the region of 3-3.5kDa, which was determined by the direct detection of bactericidal activity after SDS-PAGE.

Strain JW3 was isolated from commercial Swiss cheese products and identified as a bacteriocin producer. Lactococcus lactis JW3 showed a broad spectrum of activity against most of the non-pathogenic and pathogenic microorganisms tested by the modified deferred method. Lacticin JW3 also showed a relatively broad spectrum of activity against non-pathogenic and pathogenic microorganisms as assessed using the spot-on-lawn method. It demonstrated a typical bactericidal mode of inhibition against Leuconostoc mesenteroides KCCM 11324.

pH 및 온도에 동시에 민감한 하이드로젤을 maleilated chitosan과 N-isopropylacrylamide를 이용하여 합성하였으며, 합성된 하이드로젤은 NIPAAm의 양에 관계없이 비슷한 pH 민감성을 나타내었으며 부피상 전이온도는 모두 NIPAAm의 LCST의 온도인 $32^{\circ}C$부근에서 나타났다. 반면에 MC의 양을 소량 첨가한 하이드로젤은 그만큼 pH의 민감성이 떨어졌다. lysozyme에 의한 MC의 생분해는 2시간이내에 80%가 분해되었다. 또한 indomethacin을 이용한 약물방출 실험에서는 제조된 하이드젤이 방출제어형 약물전달체로서 좋은 결과를 보인 제형으로 관찰되었다.

A fiber optic oxygen sensor has been employed to monitor the concentrations of dissolved oxygen in a bioreactor. The characteristics of fiber optic oxygen sensor was investigated, e.g. the dependency of agitation rate on the oxygen measurement and also the dependence of temperature on the performance of fiber optic oxygen sensor etc. We have also applied to monitor the concentrations of dissoved oxygen in real cultivation processes by using the fiber optic oxygens sensor. The fiber optic oxygen sensor can be applied to measure the concentration of metabolites by immobilizing some enzymes, e.g. glucose oxidase and also employed for the environmental technology.

The highly efficient photoelectric conversion of chlorophyll a (Chl a) monolayers and multilayers was investigated. Using the Langmuir-Blodgett (LB) technique, Chl a monolayers and multilayers were fabricated onto optically transparent electrode, such as ITO glass. The photocurrent could be observed according to the light illumination. The action spectrum of the Chl a LB films was well consistent with its absorption spectrum. The possible application of the proposed system as a constituent of the artificial color recognition device was suggested.

Enantiomerically pure hydroxycarboxylic acids have great potential as chiral building blocks in fine chemicals due to their easily modificable two functional groups. Microbial polyhydroxyalkanoates (PHAs) can have more than one hundred of hydroxycarboxylic acids as monomeric constituents by growing cells under various conditions. All of the monomeric constituents are enantiomerically pure in (R)-conformation if there is a chiral center. Therefore, efficient production of enantiomerically pure hydroxycarboxylic acids by degrading PHAs is possible. In this presentation, we report on the development of a novel method for the preparation of (R)-hydroxybutyric acid by in vivo depolymerization of Polyhydroxybutyrae.

The invasion genes from Salmonella typhimurium were identified by the construction of a cosmid library and subcloning genes into a plasmid vector, pGEM-7Z. The 4.65 kb fragment of the invasion-conferring genomic region of the subclone, pSV6235 was sequenced in both direction. The three open reading frames, which were located at downstream of a promoter region, were designated as sir (Salmonella invasion region)A coding for the 36 amino acids, sirB coding for the 132 amino acids and sirC for the 82 amino acids, respectively. Interesingly, the genomic region of pSV6235 was highly homologous to Yersinia enterocolitica genomic DNA for a high pathogenicity island and Salmonella enteritidis insertion element IS1351 and IS200 DNA. These results show that there could be a significant relationship between S. typhimurium, Y. enterocolitica and S. enteritidis with respect to horizontal evolution process and acquisition of virulence determinants by means of transposon, plasmid or bacteriophage.

Sphingoliped 대사의 율속효소인 serine palmitoyl transferase(SPT)와 acid ceramidase의 연구를 위하여 인간의 SPTII 유전자와 acid ceramidase 유전자의 5‘-upstream region을 얻었다. 사람의 대장암세포인 HT29 cell로부터 genomic DNA를 얻고 GenomeWalker kit를 이용하였으며 2690bp의 SPTII promoter와 2028bp 및 1034bp의 acid ceramidase promoter의 fragment들을 얻을 수 있었다. 이들 DNA 조각들을 T7Blue vector에 subcloning하여 sequencing하였으며 이들이 사람의 SPTII 및 acid ceramidase gene의 5’-upstream region임을 확인하였다. 동물세포에서의 promoter activity를 측정하기위하여 firefly luciferase를 reporter gene으로 하는 pGL2-enhancer vector와 pGL2-basic vector에 subcloning하였으며 pRL-TK vctor와 함께 HT29 cell 및 HepG2 cell에 cotransfection 시킨 후 luciferaseg활성을 측정한 결과 같은 양의 DNA로는 사람의 SPTII promoter와 acid ceramidase promoter는 pRL-TK에 비하여 transfection efficiency가 아주 낮았으며 promoter 연구를 위하여는 pRL-TK vector의 양을 1/100으로 줄이는 것이 적당하였으며 HT29 cell보다는 HepG2 cell에 더 높은 발현율을 보였다.

In human chromosome, a short sequence of DNA has been repeated a number of times. These repeats are called variable number of tandem repeat(VNTR) or short tandem repeat(STR) which has short repeat core. VNTR and STR are used in the field of forensic science, evolution, and anthropology. In this work, we examined allele frequencies of 3 VNTR(YNZ22, NeuR, D21S11) and one STR(Humth01) in a Korean population sample by polymerase chain reaction(PCR) followed by high-resolution polyacrylamide gelelectrophoresis(PAGE) with silver staining. Subsequently, the polymorphism information content(PIC) was calculated : the highest PIC was observed for the NeuR locus(0.95680) and lowest for the Humth01 locus(0.75809).

30 kinds of natural products were considered for developing natural antioxidants by improved D.O. analyzing method including simple calculation of Area Under Curve. Several natural products such as Cimicifuge Rhizoma, Epimedii Herba, Atractylodis Rhizoma Alba, Acori Graminei Rhizoma, Mori Cortex Radicis, Aurantii Nobilis Pericarpium were bore nearly same antioxidant effects compared to synthetic powerful antioxidant BHT and also expressed powerful antioxidant effect than ${\beta}-carotene$ such as Eucommiae Cortex, Cinnamomi Cortex, Angelicae Gigantis Radix, Lycii Furctus, Acanthopanacis Cortex, Sophorae Radix, Paeoniae Radix, Geranii Herba. Another method of DPPH was performed for searching natural antioxidant from natural product. Sophorae Radix, Puerariae Radix, Aurantii Nobilis Pericarpium, Atractylodis Rhizoma Alba, Acori Graminei Rhizoma, Corni Fructus, Angelicae Gigantis Radix, Paeoniae Radix were carried higher antioxidant capacity than ${\beta}-carotene$ by DPPH method.

DNA chip containing 207 E. coli genes related to important metabolisms such as (TCA cycle, glycolysis, fermentation and etc) were used to carry out a comprehensive investigation of the change in metabolism and physiology during high cell density culture of E. coli by fed-batch cultivation.