Earticle

The theoretical research of olfaction began about a hundred years ago and the electrophysiological expermental techniques have been used for the olfaction research from 1950's. However, olfaction has not been studied so much as other senses. Recently interest in the offaction mereases for its industrial applications. We descenbe the companson of vertibrate and insect olfactory organs, smell perception mechanism, olfactory signaing transduction, and industnal applications f olfactory system, it is expected that the vanous ongeing researches on the olfactory system will contribute to sensor and scent industnes.

An investigation was made to develop new biofilm medium which could be applied to the Sequencing Batch Biofilm Reactor(SBBR) system for enhanced nutrient removal. 21 kinds of polyurethane media were tested fro adhesion ability for nitrifying bacteria. Nitrification rates were also tested by introducing synthetic wastewater containing ammonium-nitrogen to reactors with biofilm media. It was found that Z96-06 medium had higher selective adhension ability for nitrifying bacteria than the other biofilm media. The nitrification rate was 2.21 mg {{{{ { NH}`_{4 } ^{ +} }}}}-N /Lhg MLSS when we operated the SBBR system containing Z96-06. Nitrification rate of the SBBR system increased approximately by 30% compared with that of the Sequencing Batch Reactor(SBR) system which did not contain biological carrier.

In the production of a low cost bacterial insecticide, it is important to produce a high spore concentration using low price substrates. Experiments were carried out to investigate the effects of the addition of mineral salts and glucose, and of dissolved oxygen concentration on the cell growth and spore formation of Bacillus thuringiensis var aizawai using a cheap wheat and soybean meal in the batch culture. The maximum viable cell number was 1.2109 CFU/mL at 12 hr culture and spore yield was 54.2% at 74 hr culture using an industrial medium containing 20 g/L wheat meal and 30 g/L soybean meal under 1.0 vvm aeration and 200 rpm agitation. The cell growth and the spore formation were not enhanced by the addition of mineral salts in industrial medium, whereas th addition of 10g/L glucose decreased the cell growth and spore formation. We could obtain a maximum viable cell number of 2.2109 CFU/mL and spore number of 1.9109 CFU/mL at the dissolved oxygen concentration of 60% of saturation. The spore concentration was enhanced approximately by 2 times as compared to the dissolved oxygen concentration of 50%. In the bench-scale culture, the maximum viable cell and spore number were 2.5109 CFU/mL, and 2.2109 CFU/mL, respectively under 1.0 vvm aeration and 400 rpm agitation. The spore yield was 88% based on the maximum viable cell number. As a result, it was confirmed that the production of high spore concentration could be obtained by a bench-scale culture using an industrial medium.

Hantaviruses are rodent hosts-borne viruses belonging to the family Bunyaviridae, and are etiologic agents for two acute diseases, i.e., Haemorrhagic Fever with Renal Syndrome (HFRS) and Hantavirus Pulmonary Syndrome (HPS). There have been a lot of reports on prophylactic vaccines and diagnostics for the diseases, but most of viral antigens have been prepared by eukaryotic cell culture. Nucleocapsid proteins of Hantaviruses are known as the major viral antigens. Thereby, we prepared nucleocapsid genes of Hantaan virus and Seoul virus by RT-PCR and cloned into plasmid vectors, pET-3a and pKK223-3. Both genes were expressed in Escherichia coli with higher expression level of Seoul viral nucleocapsid protein compared to that of Hantaan in pET-3a. Hantaan viral gene was expressed much higher level in plasmid pET-3a that in pKK223-3. About 30% of expressed nucleocapsid protein was soluble and the rest was remained in insoluble fraction.

Hantaan virus belonging to the genus Hantavirus and family Bunyaviridae causes an acute severe illness of human, Haemorrhagic Fever with Renal Syndrome (HFRS). It is a rodent host-borne pathogen and distributed in Asia and Eastern Europe. Hantaviruses have three major antigens, i.e., G1, G2 glycoproteins and nucleocapsid protein (N). Among them, nucleocapsid protein was reported to be the most invaluable antigen as for diagnosis. We have cloned and expressed Hantaan viral nucleocapsid gene in E. coli BL21(DE3). In this study, we have tried to purify the nucleocapsid protein produced by recombinant E. coli, and could attained a purity of >90% by anti-N monoclonal antibody-coupled immunoaffinity chromatography or phenyl sepharose hydrophobic interaction chromatography.

Viruses belonging to the Hantavirus genus cause two acute severe illness in humans, i.e., Haemorrhagic Fever with Renal Syndrome (HFRS) and Hantavirus Pulmonary Syndrome(HPS). Among them, Hantaan virus is one of the most important viruses causing HFRS. Recombinant expression vectors, pKK-NP and pET-NP, with Hantaan viral nucleocapsid gene were constructed, and used to transform Eschericia coli BL21(DE3). Stability of the vectors in the host strain, and effects of some environmental conditions on the expression of nucleocapsid gene were studied. Expression vector, pKK-NP, was very unstable, and the expression level of nucleocapsid gene was very low compared to that of pET-NP. BL21(pET-NP) produced about 100 mg of N protein per liter of culture broth. Induction time did not show any significant difference on the expression level of nucleocapsid gen and cell growth. BL21(pET-NP) culture at 35℃ showed a little higher expression level than at 30℃ during growth phase, but reached to the same level at stationary phase. Total expression level was proportional to supplemented glucose concentration of media up to 0.5% along with cell growth, but expression level per unit cell mass was inversely proportional to glucose concentration and maximal when glucose was not supplemented at all.

This study was carried out particularly focusing on he antitumor effect in combination with 5-fluorocytosine(5-FC), antifungal agent, and extracellular cytosine deaminase from Chromobacterium violaceum YK 391 against U-937, K-562 and SNU-C4 cells. While the addition of 10ug/100ul of anticancer agent, 5-fluorouracil(5-FU), to U-937, K-562 and SNU-C4 caused the decrease of proliferation 90%, 75% and 93% respectively, the addition of 20ug /100ul of the extracellular cytosine deaminase and 10ug /100ul of antifungal agent 5-FC caused the decrease of proliferation 80%, 70% and 90%, respectively. These results, therefore, reveal that this enzyme has the similar clinical effect for considering of adjuvant antitumor effect. From the above results, the treatment of 5-FC and the cytosine deaminase was very effective and showed the possibility to remove side effects which easily occur by the treatment of 5-FU only. An extracellular cytosine deaminase.

In both 7L and 20L fermentor experiments the level of dissolved oxygen (D.O) strongly affected growth and PHBV production of Azotobacter vinelandii UWD. A higher D.O. increased carbon substrate consumption rate and cell growth rate with a similar residual biomass production. However, a lower D.O. was a much better condition for PHBV production. In a 20L fermentor experiments controlled at 5% D.O. cell growth rate was about twice faster(0.555 hr-1 and 0.260 hr-1 at the acid and the glucose phase, respectively) with an equal amount(4.5 g/L) of residual biomass production. However, PHBV content in the cell(62.3 wt%) increased 17.3 times at 1% D.O.

5L 발효기에서 글루코스의 투입방법과 배지의 초기 글루코스 농도에 따른 유가식 배양에 의한 S. marcescens의 증식에 미치는 영향에 대해서 연구하였다. 발효기 조건을 일정하게 유지시켜준 각 유가식 배양에서의 최종란체량이 회분식배양에 비해서 훨씬 높았으며, 유기식 배양종에서도 기질인 글루코스의 공급을 달리하였을 때, 최종균체량에서 차이를 나타내었다. 회분식 배양과 유가식 배양중에서 글루코스의 일정비율공급밤법, 글루코스 결핍상태에서의 일정비율공급방법 그리고 지수적 공급방법의 최종균체량은 1.40, 5.07, 6.93 및 7.60g/L 이고, 그 때의 배양시간은 40, 41. 24 및 40hr이었다. 그리고 균체 생산성은 0.035, 0.124, 0.239 및 0.1909/L.h 이었다. 최종 균체량만을 가지고 비교하였올 경우에는 유가식 배양줌에서 지수적 공급방법이 가장 효율적이지만 당위시간당 균체 생산성에서는 글루코스 결핍 상태에서의 일정비율공급방법이 1.52배 정도 높게 나타냈으므로 가장 효과적인 배양 방법임을 알 수 있다. 용존산소와 글루코스 루입시가의 상관관계에서 나타난 바와 같이 글루코스 결핍상태에서 일정비율로 공급한 방법과 지수적으로 공급한 방법에서 좋은 일치를 보여주었다

본 연구는 초임계 이산화탄소를 이용한 섬유소의 효소 가수분해에서 효소의 안정성 및 반응조건에 관한 연구를 수행하였다. 초임계 이산화탄소에서 cellulase의 안정성에 대한 실헝결과 압력에 대한 영향에서는 80 atm에서 160 atm까지 효소의 안정성이 유지되었으며 200 atm 에서는 약간 감소하였다. 반응시간의 경우에는 150분까지 효소의 활성이 그대로 유지되었으며, 온도는 상압에서의 최적온도인 50℃ 까지는 효소의 활성이 유지되었으나 그 이상의 온도에서는 효소의 변성에 의하여 활성이 감소하였다, 초임계 이산화탄소에서 cellulose를 120 atm과 50℃ 애서 90분간 cellulase로 가수분해 반응을 수행한 결과 20 g/L 의 Avicel이 완전히 가수분해되어 100% 수율의 glucose블 얻을 수 있었으며, 이는 상압에서 보다 glucose 수율이 1.5배 증가한 결과이다. 반면에 cellulose fiber인 경우는 상압에서 보마 1.9배 증가하였다. 통일 조건에서 Avicel의 농드를 60 g/L로 한 경우에는 glucose 수율이 상압에서 보다 1.2배 증가하였다

Experimental studies were carried out on the use of supercritical fluid in enzymatic hydrolysis of cellulose. In order to effectively perform the hydrolysis the enzyme has to maintain stability and activity in the supercritical carbon dioxide solvent. In the experiment it was found that the stability of cellulase was maintained up to 160 atm for 90 min at 50℃. In the enzymatic hydrolysis of cellulose at supercritical conditions using carbon dioxide at 80 atm and 50℃ for 90 min, the results showed that glucose yield was 100%, which was 1.5 times as compared to that in atmospheric condition when the substrate (Avicel) concentration was 20 g/L. For the substrate concentration of 60 g/L, the glucose yield was increased by 1.2 times as compared to that in atmospheric condition.

고정화한 P. phosphoreum의 biolummescence악 독생물질과의 상관관계를 이용한 수계의 독성물질 momtonng 가능성을 조사하였다. P. phosphoreum을 이용한 수계의 독성물질 momtonng을 위한 적정 세포 농도는 최대 biolummescence intensnty를 보인 O.D660 1.0~1.2였으며 이 농도외 세포를 2.5% NaCl 용액에 희석하여 5.0%의 alginateo에 고정화 했을 때 biolummescence 유지도까 가장 좋았다. 이러한 방법으로 monitoring이 가능한 수계의 pH는 6.0~8.0이었다. Free cell과 고정화 세포로 CdCI2.H2O,PbCI2,Phenol 그리고 mtrophenol에 대한 bioluminescence를 조사하였을 때 고정화 방법에 의해 독성물질에 대한 민감성0] 증가되었다. 고정화 세포외 경우 각 독성 물질에 따라 20분 이내에 0.01~1.50ppm이하까지 mornitoring이 가능하였다. 뿐만 아니라 speciflC blolurninescence reclucLion rate, biolummescence mtenslLy의 ratio 그리고 Gamma value로 분석함으로서 독성물질의 농도와 biolummescence의 변화를 직선의 관계로 나타낼 수 있어 독성 물질의 농도 예측이 가능하다.

The relationship between bioluminescence of immobilized Photobacterium phosphoreum and toxic substances was investigated to monitor toxic substances in aqueous solution. The sodium alginate was used as an immobilization matrix. A bioluminescence intensity was maximum when OD660 for cell concentration were between 1.0 and 1.2 and the biolumescence was stable at the pH range of between 6.0 and 8.0. The optimum concentration of alginate for immobilization was found to be 5.0%(w/v) in which dilution was carried out with 2.5%(w/v) NaCl solution that is an optimum environmental condition for the growth of P. phosphoreum. The bioluminescence intensity responded against the toxic substances was proportional to the concentration and a regression curve were established with linearity by using specific bioluminescence reduction rate and Gamma values. It was also found that the response was very rapid and sensitive. The response with such rapidity and sensitivity is a very important factor for the real time monitoring. The immobilized cells showed higher sensitive response to the toxic substances than free cells.

Studies were made to investigate effects of the scale-up of stirred tank bioreactors on cell growth and alkaloids production for suspension cultures of Eschscholtzia californica. In the 1.5 L STR, cell lysis was observed at 110 rpm or higher agitation speed. The agitation speed of 30 L STR was 43.7 rpm to maintain the same shear stress developed in 1.5 L STR of 100 rpm. As a result of scale-up from 1.5 L to 30 L STR, the specific growth rate was decreased from 0.12 to 0.07 day-1. The alkaloids productivity was also decreased from 0.24 to 0.14 mg/L-day. Changes of mixing performance and oxygen transfer were studied to explain the decrease of cell growth and alkaloids production. Decreased oxygen transfer rate coefficient(KLa) and increased mixing time by the scale-up was observed at various aeration rates.

과당계열 배당체인 메틸프룩토시드를 반응출발문질로하여 과당계열 당 지방산 에스테르인 메틸프룩토시드 올레산 모노 및 디에스테르를 효소적 용매 및 무용매공정에 의하여 합성하였다. 용매공정으로부터 머틸프룩토시드 올레산 모노에스테르를 합성한 결과, 최적 상용용매로는 2-메틸-2-프로관올이 우수하게 나타났으며 매틸프룩토시드의 용해도는 애서 400g/L이상에 상당하였다. 최적반응조건을 검토한 결과, 에틸프룩토시드 올레산의 최적올비 3.1, 에틸프룩토시드의 최적 초기농도 200g/L, Novozym 435의 최적 촉매함량 1%(w!v), 반응온도 60℃에서 반응시간 8시간만에 70%의 평 형 젠화율애 도달하였으며 12.6g/L-hr의 생산성으로 매렐프룩토시드 올레산 모노에스테프 블 생싼할 수 았었다. 그리고, 무용매공정에서 메틸프룩토시드 올레산 디에스터르를 합생한 결과, 메틸프룩토시드와 올레산의 최적돌에 l.2, Novozym 435의 최적 촉매함령, 10%(w/w), 반응온도 , 진공도 20 - 200mmHg 에서 반응시간 10시간만에 95%이상의 전화율로 140g;L-hr의 생산성으로 메틸프룩토시드 올레산 디에스테르를 생산할 수 있었다.

Enzymatic synthesis of fructose-based sugar fatty acid esters, such as methyl fructoside oleic acid mono and diester, was investigated using methyl fructoside as a sugar starting material. For the production of methyl fructoside fatty acid monoester by solvent process, 2-methyl 2-propanol was found to be a god reaction medium resulting a higher yield and productivity due to its high sugar solubility. The yield and productivity of methyl fructoside oleic acid monoester were 70% and 12.6g/L-hr, respectively, when molar ratio of methyl fructoside, initial concentration of methyl fructoside, enzyme(Novozym 435) content, and reaction temperature were 3:1, 200g/L, 1%(w/v), and 60℃, respectively. Methyl fructoside oleic acid diester was prepared by lipase-catalyzed diacylation of methyl fructoside and oleic acid in the solvent-free process. Maximum yield of 98% and productivity of 140g/L-hr were achieved when molar ratio(methyl fructoside and oleic acid) of 1:2 enzyme content of 10%(w/v) and reaction temperature of were applied for the operating conditions under a reduced pressure of 20∼200 mmHg.

The effect of glycyrrhizin on melanoma cells was investigated. DNA fragmentation in cultured melanoma cells was promoted by the addition of glycyrrhizin in a dose dependent manner. Administration(i.m.) of glycyrrhizin to Mel/ret transgenic mice resulted in apoptosis induction, reduction of mitochondrial transmembrane potential in melanoma cells. Decreased B220+ B cells were recovered by the treatment of glycyrrhizin in splenocytes and mesenteric lymph node cells, while Thy-1+ T cells were not influenced. Results suggested that glycyrrhizin acted as an inducer of apoptosis of melanoma cells and an immuno-potentiator via recovered B lymphocyte population in Mel/ret transgenic mice.

To develop a large scale countercurrent single pass heat exchanger for continuous transportation and sterilization of soybean paste, microflora and color value of sterilized soybean paste were examined at various sterile condition. Aerobes, anaerobes, molds, yeasts and lactic acid bacteria were 5.1 x 107 CFU/g, 7.1 x 107 CFU/g, 2.6 x 105 CFU/g, 4.3 x 106 CFU/g, 1.3 x 107 CFU/g, respectively in raw soybean paste. In gold band ampoule test, aerobes and anaerobes of soybean paste were viable up to 90℃, but become unviable at 100℃. Molds decreased rapidly and yeasts decreased slowly from 70℃. Lactic acid bacteria were unviable at 60℃ within 10 min. In color test, Hunter L, a, and b values of soybean paste were 50.2, +5.6, and +17.8, respectively. After heating in polyethylene film bag at 80℃, Hunter values were not so much changed and become 50.2, +4.7, and +19.7, respectively. The micorflora and color of soybean paste sterilized in a large scale heat exchanger system resulted in very similar to those of gold band ampoule and polyethylene film bag by effective heat transfer.

An이ew식과 Aiba삭을 조합하여 에탄올 생산단주인 Saccharomyces cerevisiae ATCC 24858의 비성장속도를 당농도와 에탄올 농도의 함수로 표현하였다. 기침의 저해영향을 받지 않는 최대 당농도 은 150 g/L이며 기질의 저해영향은 기질농도 S와 S-Smax항의 함수로 표현되었다. 최대 비성장 속도 umax[?][?][?]0.49hr-1,Monod[?][?][?][?][?][?]K1는 19 g/L, Andrew식의 기질저해상수 K1는 139 g/L이였다. 또한 비성장속도에 영향을 마치지 앓는 최대알콜농도 Pm이 존재하였으며 그 값은 2 g/L 이였다. 따라서 Aiba식에서 비성장속도에 영향을 미치는 에탄올 농도항은 P-Pm으보 표현되었다. 본 연구의 알코올생산균주에 대한 비성장속도의 완성된 수식은 디음과 같으며 이 수식에 위한 계산값은 평균오차 6% 내외의 범위에서 실험값과 일치하였다

The mathematical model of specific growth rate of Saccharomyces cerevisiae ATCC 24858 is proposed as a function of sugar and ethanol concentrations by the combination of Andrew's equation and Aiba's equation. The maximum concentration of sugar Sm, which was the highest concentration of sugar not having any effect on the growth inhibition, was 150 g/L and the substrate inhibition was expressed as a function of (S-Sm). The maximum specific growth inhibition, was 150 g/L and the substrate inhibition was expressed as a function of (S-Sm). The maximum specific growth rate um, Monod's constant Ks, and Andrew's inhibition constant KI were 0.49 hr-1, 19 g/L, and 139 g/L, respectively. The maximum ethanol concentration, Pm, which did not show any inhibition effect on the specific growth rate was found to be 2 g/L. Therefore, the ethanol inhibition was represented as a function of (P-Pm). The final mathematical model for the specific growth rate of the microorganism in this work is proposed as the following. And the average percent of errors between the calculated specific growth rate and the experimental values was 5.96%.