Earticle

Biofiltration is a bioprocess treating volatile organic compounds (VOCs) in order to convert the VOCs to harmless products. This review on biofiltration is intended to provide an engineering concept such as removal efficiency, maximum load, elimination capacity and so on. Besides, modeling concept of biofilter is also supplied for designing biofilter system. Quantitative data generated in our research group is shown to explain the engineering concept as well as the modeling idea.

Immunosensors are of great interest because of their potential utility as specific, simple, label-free, direct detection means and provision of reduction in size, cost and time of analysis comparing with conventional immunoassay. In the last two decades, many reports have been published on the use of immunosensors for a wide range of applications to clinical diagnostics, pharmaceutical chemistry, environmental monitoring, biotechnology and food industries. There are also numerous transduction techniques developed such as electrochemical techniques, piezoelectric crystal, and surface plasmon resonance receiving much attention for the direct monitoring of immune reactions at solid surfaces. In this article, the principles, characteristics, structures, functions and clinical applications of immunosensors were reviewed

산가수분해법과 칼럼크로마토그라피법을 이용하여 레반 올리고당과 저분자량 레반을 생산하였다. 레반에 대한 산가수분해반응은 시간의존적으로 비례적으로 진행되었으며, 다른 미생물 유래의 레반을 사용시에도 동일한 결과를 나타내었다. 레반 올리고당의 제조를 위한 최적화된 조건은 5% 레반을 0.38 M 황산, 95oC, 4분간 처리하였을 때 였으며, 최종생산물로 중합도 3-6의 레반 올리고당을 얻었다. 생성된 레반 올리고당을 탄소원으로 이용하여 두 젖산 생성균주(Lactobacillus plantarum KCTC 3104와 Pediococcus pentosaceus KCTC 3507)의 생장배지에 첨가하고 레반 자체를 첨가한 배지와 비교시, 뚜렷한 생육촉진효과와 pH 감소효과가 나타났다.

Levan oligosaccharides and low molecular weight levan were produced from levan by acid hydrolysis and following column chromatography. Levan hydrolysis was progressed proportionally as increased incubation time. In terms of levan hdrolysis reaction, no differences were found from the sources of levan. Optimum hydrolysis conditions for the formation of levan oligosaccharides were, 0.38 M H2SO4; and incubation at 95oC for 4 min. The purified products were determined as the mixture of oligosaccharides (degree of polymerization (DP) of 3-6). Two of lactic acid-producing bacteria, Lactobacillus plantarum KCTC 3104 and Pediococcus pentosaceus KCTC 3507, were studied in vitro for their ability to metabolize levan oligosaccharides. Apparently, the growth of both cells were increased by levan oligosaccharide diet, compared with those of levan diets, suggesting that levan oligosaccharides may be beneficial in selectively growth of lactic acid-producing bacteria.

기포탑 회분식 xanthan 발효에서 xanthan 생성속도와 분자량에 영향을 미치는 산소전달제한을 피하기 위하여 과산화수소수를 산소공급 보조수단으로 사용하였다. 과산화수소수 주입은 xanthan 생성을 가역적으로 저해하였다. 반면에 직경 8 mm 유리구슬 유동화 입자는 기-액 산소전달속도를 증가시켜서 기-액-고 삼상유동층 생물반응기에서의 xanthan 발효는 기포탑 발효에 비하여 높은 단위균체량 당 xanthan 생성속도 및 점도수율 그리고 반응기 공간-시간 수율(space-time yield)을 보였다.

The decomposition of hydrogen peroxide was used for supplementing the oxygen during batch xanthan fermentations in a bubble column bioreactor in order to escape the oxygen transfer limitation that occurred at the high viscosity of culture broths. The xanthan production, however, was inhibited reversibly by dosing hydrogen peroxide. On the other hand, fluidized particles of glass beads with 8 mm diameter led to high gas-liquid oxygen transfer rates in three-phase fluidized beds, which resulted in higher space-time yields of the xanthan production compared to in the bubble column bioreactors.

초임계 이산화탄소를 이용한 유자과피로 부터의 휘발성 정유성분의 추출은 entrainer를 사용하지 않을 경우 압력 13.8 MPa, 온도 40℃에서 786.61 mg/sample 30 g이 추출되었으며 entrainer를 0.4 mL/min을 첨가하였을 경우 압력이 13.8 MPa, 온도 40℃에서 5416.64 mg/sample 30 g이 추출되어 약 7배의 추 출량의 증가를 볼 수 있었다. 이는 보조용매를 첨가함으로서 초임계 이산화탄소의 유자과피의 휘발성 정유성분에 대한 용해력을 증가시키는데 큰 영향을 미치는 것으로 알 수 있다.

The extraction of volatile essential oil using supercritical carbon dioxide with 2%(v/v) ethanol and non-ethanol was performed in a semi-continuous flow extractor at the range of pressures and temperatures 6.9 to 17.2 MPa and 30 to 45oC, respectively. When ethanol was added to the co-solvent, the solubility of volatile essential oils was increased up to 90% over the neat CO2 value. The most efficiency of extraction of the voilatile essential oils was achieved at 13.8 ㎫ and 40oC by supercritical carbon dioxide with entrainer from Citrus junos peel.

Poly-lysine이 tagging된 hEGF와 angiogenin(6L10E5A)의 융합단백질의 고체상 재접힘이 heparin-Sepharose column에서 수행되었을 때, untagging 단백질(E5A)의 기존의 액상 재접힘방법과 비교하여 재접힘 수율은 약 13배 정도 증가하였다. 게다가 poly-lysine tagging된angiogenin은 heparin에 친화도를 높여주므로 2.5배에서 3배 정도의 흡착 수율이 증가한다. 재접힘 수율은 고체상 반응으로 인해 높은 재현성을 보였다. 재접힘 공정시간은 대략 8배 단축되었다. 고체상 재접힘된 단백질은 자신의 생물학적 역가를 유지하였다. 따라서 이연구는 고체상 재접힘 방법이 분자간의 상호작용을 억제하여 응집현상을 현저히 줄였기 때문에 기인한 결과로 생각된다. 따라서 응집으로 인한 재접힘 수율이 낮은 단백질의 재접힘공정에 고체상 재접힘 공정을 사용하면 높은 재접힘 수율을 얻을 수 있다.

A fusion protein, consisting of a human epidermal growth factor as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as an inclusion body in recombinant E. coli, yet when the conventional solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably because of the opposite surface charge resulting from the vastly different pI values of each domain. Accordingly the solid-phase refolding process, which exploits the ionic interactions between a solid matrix and the protein, was tried, however the ionic binding yield was also very low regardless of the resins and pH conditions used. Therefore, to provide a higher affinity toward the solid matrix, six lysine residues were tagged to the N-terminus of the hEGF domain. When cation exchange resins, such as heparin- or CM-Sepharose, were used as the matrix, the adsorption capacity increased 2.5～3-fold and the subsequent refolding yield increased nearly 15-fold compared to the conventional process. A similar result was also obtained when an Ni-NTA metal affinity resin was used.

본 연구에서는 화학무기, 농약 등에 사용되는 신경독성물질인 유기인화합물의 측정을 위하여 유기인화합물의 효소반응 저해작용을 이용한 광바이오센서장치의 시제품을 제작하였다. 효소반응을 위하여 효소로는 신경세포의 필수효소인 acetylcholinesterase, 기질로는 acetylthiocholine iodide을 사용하였으며 효소반응의 저해제인 유기인화합물로는 paraoxon을 사용하였다. 센서의 측정원리는 유기인화합물에 의해 저해된 효소반응정도를 효소반응의 생성물인 아세트산의 정량적 측정으로 분석하였으며, pH에 의하여 최대 흡광파장의 변화가 일어나는 litmus를 사용하여 흡광도 측정으로 아세트산의 정량분석을 수행하였다. 광바이오센서 시제품의 제작은 광원으로 고휘도 LED와 광세기 측정을 위한 photodiode로 구성하였으며, 제작된 센서를 이용한 실험결과로부터 0 ppm에서 2 ppm의 paraoxon 농도에서 구성된 센서시스템의 선형적 신호 변화를 관찰하였다. 이상의 실험결과로부터 광바이오센서시제품은 2분의 반응시간으로 신속하고 정확한 유기인화합물의 정량분석이 가능함을 확인하였다.

In this study, a prototype fiber-optic biosensor was fabricated using the inhibition of enzyme reaction by organophosphorus compounds to detect organophosphorus compounds, which is nervous toxic material and is used as chemical weapon and pesticide. Enzyme, substrate, and inhibitor for enzyme reaction were acetylcholinesterase (key enzyme in nervous cell), acetylthiocholine iodide, and paraoxon (a kind of organophosphorus compounds), respectively. The detection principle of sensor is the detection of enzyme reaction inhibited by organophosphorus compounds by the quantitative measurement of acetic acid, which was achieved by absorbance measurement using litmus solution that maximum absorbance band is changed by pH. To fabricate prototype fiber-optic biosensor, high bright LED and photodiode was used as light source and light intensity detector, respectively. From the experimental results using a prototype biosensor, the linear change of sensor signal was obtained in a range of 0-2 ppm inhibitor concentrations. From these results, it was verified that the quantitative measurement of organophosphorus compounds could be achieved fast (within 2 minutes) and accurately by a prototype fiber-optic biosensor.

본 연구에서는 유도성 promoter인 GAL1과 상시성 promoter인 GPD와 ADH1 promoter 하류에 사람 ferritin H-chain 유전자(hfH) 및 사람 ferritin L-chain 유전자(hfL)를 연결하여 재조합 plasmid를 구축하고 이들을 S. cerevisiae 2805에 형질 전환시켜 외래 유전자를 성공적으로 발현시켰다. Ferritin 발현에 미치는 promoter의 영향을 비교한 바, 이 두 단백질 생산에 있어서 GAL1 promoter가 GPD나 ADH1 promoter 보다 더 효율적이었다. GAL1 promoter를 사용한 형질 전환체(YG-H와 YG-L)에서 H-chain의 발현율은 전체 수용성 단백질 중 4.5%에 해당하였고, L-chain의 발현율은 9.8%에 이르렀다. 각 균주에서 발현된 H 및 L subunit들은 비변성 젤을 사용한 전기 영동의 결과 대장균에서 생산된 재조합 단백질과 마찬가지로 자발적으로 holoprotein으로 조합되어졌다. 재조합 H-와 L-ferritin들은 단백질 내공에 철을 축적할 수 있음이 Prussian blue 염색으로 확인되었다. 그리고 효모 세포를 10 mM ferric citrate를 함유한 배지에서 배양했을 때, H-ferritin과 L-ferritin을 생산하는 재조합 효모 균주에 있어서의 철의 농도는 각각 174.9 μg per gram(dry cell weight)과 148.8 μg per gram(dry cell weight)이었고 야생형 효모 균주에 있어서의 철의 농도는 49.4 μg per gram(dry cell weight)이었다. 이것은 사람 ferritin 유전자를 효모 균주에 발현시킴으로써 효모의 철 함량이 증진되었음을 유추하는 결과이다.

Human ferritin H- and L-chain genes(hfH and hfL) were cloned into the yeast shuttle vector YEp352 with various promoters, and the vectors constructed were used to transform Saccharomyces cerevisiae 2805. Three different promoters fused to hfH and hfL were used: galactokinase 1(GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase(GPD) promoter and alcohol dehydrogenase I(ADH1) promoter. SDS-polyacrylamide gel electrophoresis and Western blotting analyses displayed expression of the introduced hfH and hfL. In the production of both ferritin H and L subunits GAL1 promoter was more effective than GPD promoter or ADH1 promoter. Ferritin H and L subunits produced in S. cerevisiae were spontaneously assembled into its holoproteins as proven on native polyacrylamide gels. Both recombinant H and L-chain ferritins were catalytically active in forming iron core. When the cells were cultured in the medium containing 10 mM ferric citrate, the cell-associated concentration of iron was 174.9 μg per gram(dry cell weight) for the recombinant yeast YG-H and 148.8 μg per gram(dry cell weight) for the recombinant yeast YG-L but was 49.4 μg per gram(dry cell weight) in the wild type, indicating that the iron contents of yeast is improved by heterologous expression of human ferritin H-chain or L-chain genes.

원유로부터 다당류를 생산하는 유산균을 분리, 선발하여 균의 배양조건을 조사하고 Lactobacillus ssp. SCU-M으로 동정, 명명된 최적배지의 탄소원은 galactose 1.5% 첨가시 553 mg/L의 EPS를 생성하였으며 whey의 이용성 검토를 위한 deproteinized whey와 lactic acidified whey를 탄소원으로 사용하였을 때 각각 639, 550 mg/L를 생성하였다. 질소원으로 yeast extract 1.5%, peptone 0.5%를 첨가한 경우 934 mg/L, 미량성분으로 MgSO4, 0.15%, K2HPO4 0.15% 첨가시 1,076 mg/L의 다당생산성을 나타내었다. 따라서 이들 결과를 종합하여 galactose 1.50%, yeast extract 1.00%, peptone 0.25%, MgSO4․7H2O 0.15%, K2HPO4 0.15%, tween 80, 0.10%를 적정 배지로 하여 pH 6.5, 배양온도 37℃에서 배양시 1,680 mg/L의 EPS를 생성하였다.

A lactic acid bacterial isolate Lactobacillus ssp. SCU-M which produces exopolysaccharide was identified and its cultural condition was investigated. The optimum culture conditions for exopolysaccharide(EPS) production of Lactobacillus ssp. SCU-M were 37oC, pH 6.5, using medium composed of 1.5% galactose, 1.0% yeast extract, 0.25% peptone, 0.15% MgSO4․7H2O, 0.15% K2HPO4 and 0.1% tween 80 in distilled water. The EPS concentration after 48 hours at the initial pH 6.5, 37oC in a flask culture was 1,680 mg/ℓ.

Astaxanthin (3,3'-dihydroxy-β,β-carotene-4-4'-dione), a natural pigment of pink to red color, is widely distributed in nature particularly in the skin layer of salmonoids and the crust of shrimp, lobster, etc. Recently, it was produced from the yeast culture of Phaffia rhodozyma. Because of its high thermal stability and antioxidant functionality, its applications can be extended into food, cosmetics, and pharmaceutical ingredient beyond the traditional feed additive. Because of its very high lipophilicity, astaxanthin has been extracted traditionally by strong organic solvents such as chloroform, petroleum ether, acetone, etc. In this study, we developed a surfactant-based solubilization system for astaxanthin, and used it to extract astaxanthin from disrupted yeast cells. Among Tween 20, Triton X-100 and SDS, Tween 20 was identified as the most suitable surfactant in terms of extraction capacity and safety. The ethylene oxide group of Tween 20 was identified as the most significant factor to increase the HLB value that determined the extraction capacity. The effects of micelle formation condition, such as the molar ratio of astaxanthin and Tween 20, pH, and ionic strength were also investigated. pH and ionic strength showed no significant effects. The optimal molar ratio between astaxanthin and Tween 20 was 1：12. Antioxidant activity of astaxanthin was higher than β-carotene and α-tocopherol. Astaxanthin in the crude extract from the yeast cell was more resistant to air and/or light degradation than pure astaxanthin, probably because of the presence of other carotenoids and lipids.

푸마르산으로부터 숙신산 생물전환을 위하여 E. faecalis RKY1을 HFBR에 고정화하여 연속생산 공정에 대한 가능성을 모색하였다. E. faecalis RKY1은 실관의 spongy 부분에 효과적으로 고정화되었으며, transverse mode로 HFBR 조업시 실관가닥이 50개 및 200개일 경우 거의 비슷한 경향을 나타냈다. 또한 배지의 공급속도를 0.25, 0.5, 1.0 mL/min로 증가시키면서 조업한 결과, 정상상태에 도달하는 시간이 각각 24,12, 9시간으로서 유속이 증가할수록 정상상태에 도달하는 시간은 단축되었지만, 반응기 내의 기질 및 생성물 분포는 그 변화가 심하였다. 실관 생물반응기를 이용하여 숙신산 생물 전환시 배지의 공급 유속이 증가할수록 숙신산 생산성은 증가하였지만 전환수율이 감소하여 미반응 푸마르산은 증가하였다. 최대 숙신산 생산성 및 이 때의 수율은 푸마르산염 농도 80 g/L 및 배지 공급 유속 2.0 mL/min일 때 17.1 g/L․hr 및 0.54 g/g이었으며, 최적 숙신산 생산성 및 이 때의 수율은 푸마르산염 농도 50 g/L 및 배지 공급 유속 1.0 mL/min일때 9.0 g/L․hr 및 0.90 g/g으로서 회분식 생물전환의 경우보 다 더 우수하였다.

Enterococcus faecalis RKY1 cells were immobilized in an asymmetric hollow fiber bioreactor for application to the continuous production of succinic acid. The media was fed into shell-side of the module using a peristaltic pump, and the products were collected through lumen-side outlet. The number of hollow fibers within the module did not affect the bioreactor efficiency in the transverse operated hollow fiber bioreactor. The steady state at the outlet of hollow fiber bioreactor was reached after 24 hr cultivation at flow rate of 0.25 mL/min, 12 hr at 0.5 mL/min, and 9 hr at 1.0 mL/min, respectively. The succinate and fumarate concentrations within the hollow fiber bioreactor, however, were as changeful as increasing the flow rate. During continuous operation with the flow rates between 0.5 and 2.0 mL, the productivity of succinate was 8.0-10.9 g/L․hr at 30 g/L fumarate, 4.9-14.9 g/L․hr at 50 g/L fumarate, and 7.2-17.1 g/L․hr at 80 g/L fumarate, respectively.

Trametes sp.에서 생산되는 laccase는 CNBr-activated Sepharose-4B(CAS4B)에 고정화 되었고, 염료의 연속적인 분해를 위하여 테스트되었다. Laccase는 CAS4B에 효율적으로 고정화되었고, CAS4B에 고정화 된 laccase는 pH, 열, 단백질 구조적인 안정성 면에서 상당히 증가하였다. CAS4B에 고정화 된 laccase의 최적 pH는 5, 온도는 40℃로서 free laccase와 비교하여 변화가 없었다. 기질로서 Reactive Blue 19를 사용하였을 때 free laccase와 고정화 laccase의 Km(μmol/mL) 값은 각각 0.34와 2.0이었고, Vmax(μmol/mL․min)값은 각각 0.12, 0.1이었다. Repeated-batch 반응에서 효소의 안정성과 높은 분해효율 만족하는 조건은 pH 5, 30℃이였다. 또한 HBT에 의한 효소의 불활성은 크게 나타나지 않았다. Packed-bed reactor에서 최적으로 운전되었을 때 100 μM Reactive Blue 19과 0.1 mM HBT가 존재하는 50 μM Acid Red 57의 연속적인 분해에서 30시간 후에도 분해 효율이 70%로 유지되었다. 고정화 laccase는 Packed-bed reactor에서 azo 염료의 연속적인 분해를 매우 안정적으로 수행하였다.

Laccase produced from Trametes sp. was immobilized on CNBr-activated Sepharose-4B (CAS4B) and tested for degradation of azo dyes. Laccase was efficiently immobilized on CAS4B. Immobilization of laccase on CAS4B increased pH, thermal and proteolytic stabilities. Optimum pH and temperature of immobilized laccase were pH 3 and 40oC, respectively as same as those of free laccase. The Km(μmol/ml) values of free and immobilized laccase for Reactive Blue 19 as the substrate were 0.34 and 2.07, respectively. Vmax(μmol/mL․min) values of them were 0.12 and 0.1, respectively. In repeated batch reactions, conditions retained high stability and degradation of dye for immobilized laccase were pH 5 and 30oC. HBT didn't decrease highly activity of immobilized laccase. Immobilized laccase was very stable for degrading dyes continuously in a packed-bed reactor containing laccase immobilized on CAS4B. For continuous degradation of 100 μM Reactive Blue 19 and 50 μM Acid Red 57 in the presence of 0.1 mM HBT under optimum conditions, immobilized laccase retained 70% of degradation ability even after 30 hours.

사상균 FJ1의 섬유소 분해효소 생산조건을 최적화하기 위해 반응표면 분석법을 이용하였다. 반응표면 분석을 위한 실험 계획법은 중심합성법 계획법을 이용하였으며, 주요 배양인자로서 탄소원의 농도, 질소원의 농도, 혼합탄소원 비율, 그리고 배양시간에 대해 조사하였다. CMCase의 경우에서는 탄소원의 농도 3.5%, 질소원 농도 0.6%, 혼합탄소원인 avicel 및 CMC의 비율 52:48, 그리고 배양시간 5.4일에 33.5 U/mL로 최적화되었고, xylanase의 생산은 3.5%, 0.8%, 54:46의 배양조건에서 5.3일에 52.6 U/mL이었고, β-glucosidase의 생산은 5.0%, 1.0%, 83:17의 배양조건에서 7일에 2.88 U/mL이었고, avicelase의 생산은 4.0%, 0.9%, 64:36의 배양조건에서 6.5일에 1.84 U/mL로 최적화되었다. 이 중에서 β-glucosidase의 생산성은 중앙값의 실험조건보다 74% 정도로 가장 높은 효율 향상을 보여 주었다. 실제로 최적 예측조건에서 실증실험을 수행한 결과 본 모델의 타당성이 입증되었다. 본 연구에서 얻은 섬유소 분해 효소 생산 조건 최적화에 관련된 자료들은 산업적으로 효소를 생산하고자 할 때 유용하게 사용될 것이다.

The production conditions of cellulolytic enzymes by a fungus, strain FJ1, were optimized using response surface analysis. The culture factors which largely affected the production of enzymes such as cultivation time, carbon source concentration, nitrogen source concentration, and composition ratio of carbon sources were employed. Optimized conditions of the factors above corresponding to each cellulolytic enzyme production were as following: CMCase production was obtained in the conditions of cultivation time of 5.4 days, carbon source concentration of 3.5%, nitrogen source concentration of 0.6%, and composition ratio of carbon sources of 52:48 (avicel:CMC), xylanase appeared in the conditions of 5.3 days, 3.5%, 0.8%, and 54:46, respectively, and β-glucosidase were 7.0 days, 5.0%, 1.0%, and 83:17, respectively, and avicelase were 6.5 days, 4.0%, 0.9%, and 64:36, respectively. The activities of CMCase, xylanase, β-glucosidase, and avicelase predicted by the response surface methodology were 33.5, 52.6, 2.88, and 1.84 U/mL, respectively, and β-glucosidase activity was enhanced up to 74% when compared to that obtained in the experimental conditions.

신체성장은 세포수가 증가하고 세포의 크기가 커지면서 신체의 크기가 증가하는 것으로서 몸을 구성하는 성분인 단백질이 증가를 수반하고 골격과 근육이 커지며 이들의 기능도 강화된다. 사람의 키가 커진다는 것은 팔과 다리의 긴 뼈가 늘어나야 하는 것인데 이와같은 성장은 호르몬의 작용에 의해서 이루어진다. 본 연구에서는 이와같은 성장에 미치는 영향을 검토하기 위해 성장호르몬 및 IGF-1의 분비촉진 여부를 확인하였다. 체외 래트 성장호르몬의 분비량은 88 - 99 ng/mL으로 대조군에 비해 약 6배 이상의 성장호르몬이 분비되었다. 또한 생체내 IGF-1 분비촉진 효과실험에서는 YGF 투여후 8시간에서 1140 ng/mL 정도로 대조군에 비해 현저하게 높은 혈중 IGF-1의 농도가 유지되었다. YGF를 장기 복용시킨 경우 실제로 대퇴부 뼈의 길이성장이 약 6% 정도 이루어졌음을 확인할 수 있었다. 따라서 본 실험을 통해 확인된 성장호르몬 및 IGF-1 분비촉진을 유발하는 식물 추출물 YGF에 대해 좀더 연구개발하여 키 성장이 저조한 어린이 및 청소년에게 신체성장에 도움이 될 수 있도룩 되었으면 한다.

The extracts of a lot of (different) plants were studied for inducing the secretion of growth hormone and IGF-1. In the cell test, it was found that the concentration of rat growth hormone was 88 - 99 ng/mL in a certain plant extract treatment group which was at least 6 times higher than the amount in control group. The concentration of IGF-1 in blood was 1,140ng/mL at 8 hours after the oral administration of YGF, which was remarkably much higher than that in control group. This meant that YGF maintained the level of IGF-1 secretion higher in the treated group. In addition, the increase of 6% in bone length was shown for the long-term oral administration of YGF. Thus, YGF seemed to affect significantly for inducing the secretion of IGF-1 which was essential for the physical growth and anti-aging.

습식 상 전환법으로 theophylline이 각인된 poly(acrylonitrile-co-acrylic acid) 막을 제조하였다. theophylline은 고분자가 공중합되는 과정에 각인시키거나(in-situ implanting, 제자리 동시 각인) 공중합된 고분자를 DMSO 용액에 녹이면서template를 섞어 각인시켰다(post implanting, 후 각인). 막의 template molecule에 대한 흡착 선택도는 응고시키는 물의 온도가 높을수록, 혼합물의 초기 농도가 높을수록 크게 나타났다. 후 각인법으로 제조된 막의 흡착선택도가 제자리 동시각인 제작법으로 제조된 막보다 흡착선택도가 우수하였다.

The theophylline imprinted membrane was prepared by a wet phase inversion method. Theophylline was implanted during copolymerization of acrylonitrile with acrylic acid or implanted in the dimethyl sulfoxide solution containing 10 wt% copolymer, P(AN-co-AA). Rolling the glass plate, on which the copolymer solution was cast, in water removed the sponge layer and thus made the membrane symmetric. The adsorption selectivity of the membrane toward template molecule was increased with the coagulation temperature of the membrane and the initial concentration of the theophylline and caffeine mixture.