• 생명과학회지
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• 제9권4호
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• pp.414-422
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• 1999

Our works performed for preparation of oligosaccharides from carrageenan, seaweed polysaccharide, and one active strain for carrageenan was isolated from sea water and identified to Pseudomonas alcaligenes. Carrageenan degrading enzyme was purified from the culture fluid of isolated strain-Pseudomonas alcaligenes JCL-43, by DEAE-Cellulose, Sephadex G-100, Q-Sepharose and CM Sepharose CL-6B column chromatography. Two enzyme-F-I, F-II- was identified this purifying process, and the molecular weight of the purified carrageenase were estimated to be 23.6kDa and 30.2kDa, respectively. The optimum pH and temperature for two carrageenase activity were 7.0 and 4$0^{\circ}C$. These enzymes were stable in the pH range of 6.0~7.5 and lower than 5$0^{\circ}C$, and required 1.5% NaCl for optimum activity. And these carragennase were inhibited by metal ions such as Cu2+, Zn2+, Hg2+, but increased by Ba2+ and Ca2+, and showed specificity on -carrageenan.

• 미생물학회지
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• 제29권5호
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• pp.296-300
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• 1991

Determent 들을 이용하여 대장균의 막에 걀합된 수소발생효소를 부분정제하였다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.183-183
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• 1994

HIV-1 protcase 를 이용한 in vitro assay system을 개발하기 위하여HIV-1 protease유전자를 Ecoli 발현 벡터를 이용하여 발현시켰다. 가능성있는 Protease유전자의 생간 및 분래를 용이하게 하기위하여 maltose binding protein 의 fusion protein을 이용하였으며 protease 의 autoprocessing을 maltose binding protein 의 polyclonal 항체로 확인하였다, 발현된 protease는 일련의 chromatography 방법 (DEAE, SE cellulose, Superose 12, Mono S) 으로 순수하게 분리되었다. 정제된 protease 는 SDS-PAGE분석으로 단일밴드를 보여주었고, 합성된 undecapeptide를 기질로 하였을때 Km 이 9.8$\mu$M 이었다. 효소 assay 를 위해 기질이 protease 에 의해 절단된 생성물을 HPLC를 사용하여 분석하였다. Protease의 억제제 탐색을 위해 유기합성한 몇개의 기질유사체와 HIV-1 증식을 억제하는 것으로 알려진 천연물의 억제정도를 조사하여 보았다. 이들 test 에 사용한 물질들은 높은 농도에서 protease 의 활성을 저해하는 것으로 보아 좋은 억제제는 아닌것으로 시료되나 본 연구를 통하여 확립된 in vitro assay system 은 추후에도 억제제 탐색을 위하여 계속 활용될 수 있을 것이다.

• Journal of Microbiology and Biotechnology
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• 제9권2호
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• pp.219-222
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• 1999

A simple sequence of membrane concentration and DEAE-Cellulose chromatography has been optimized to give a purified dextransucrase from Leuconostoc mesenteroides B-512FMCM with the highest specific activity (248.8 IU/mg protein) ever reported in high yield (overall 88.7%) for dextransucrase. When there was no sucrose in the dextransucrase and the dextran reaction digest, the dextransucrase hydrolyzed glucose from dextran. The glucose was transferred to the other glucoses from dextran and formed isomaltose and isomaltodextrin. The transglycosylation efficiency of glucose from dextran was much higher with acceptors. The dextransucrase can be used for the production of various kinds (or structures) of oligosaccharides using dextran and various acceptors with almost 100% theoretical yield.

• 한국미생물·생명공학회지
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• 제23권2호
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• pp.203-208
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• 1995

The Polyvinyl alcohol (PVA) oxidase is a key enzyme involved in degradation of PVA with PVA hydrolase. The PVA oxidase has been purified to homogeneity from the culture broth of PVA grown Pseudomonas cepacia G5Y strain by ammonium sulfate precipitation, DEAE-cellulose column chromatography, and Sephadex G-150 gel filtration. The molecular weight of the purified enzyme was estimated about 60, 000 daltons by SDS-polyacrylamid gel electrophoresis. The enzyme is most active at 45$\circ$C and at pH 8.5, and is stable below 55$\circ$C and between pH 6 and 9. The enzyme activity was strongly inhibited by Ag$^{2+}$ and Hg$^{2+}$.

• 대한의생명과학회지
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• 제10권4호
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• pp.415-420
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• 2004

Fibrinolytic substance was purified from the Wooltalikong (Phaseolus ssp.), using DEAE-cellulose chromatography, Sephadex G-150 gel-filtration, and FPLC gel-filtration. The substance has a molecular weight of 5262.70 Da as measured by MALD-TOF mass spectrometry. It has a pH optimum at pH 6.0. The fibrinolytic activity of purified substance was inhibited by EDTA and 1,10-phenanthroline and slightly decreased by PMSF and pepstatin A. It shows the maximum fibrinolytic activity at 40℃ and the substance was stable up to 50℃. The activity of the substance was increased by Zn/sup 2+/ and was totally inhibited by Hg/sup 2+/. This study revealed that Wooltalikong could be a good source of fibrinolytic products due to its small molecular size and heat-resistant ability.

• BMB Reports
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• 제36권2호
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• pp.185-189
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• 2003

Chitinase (EC 3.2.1.14) was isolated from the culture filtrate of Streptomyces sp. M-20 and purified by ammonium sulfate precipitation, DEAE-cellulose ion-exchange chromatography, and Sephadex G-100 gel filtration. No exochitinase activity was found in the culture filtrate. The molecular mass of the purified chitinase was 20 kDa, estimated by a sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and was confirmed by activity staining with Calcofluor White M2R. Chitinase was optimally active at pH of 5.0 and at $30^{\circ}C$. The enzyme was stable from pH 4 to 8, and up to $40^{\circ}C$. Among the metals and inhibitors that were tested, the $Hg^+$, $Hg^{2+}$, and p-chloromercuribenzoic acid completely inhibited the enzyme activity. The chitinase activity was high on colloidal chitin, chitotriose, and chitooligosaccharide. The purified chitinase showed antifungal activity against Botrytis cinerea, and lysozyme activity against the cell wall of Botrytis cinerea.

• Natural Product Sciences
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• 제4권2호
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• pp.88-90
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• 1998

An acid polysaccharide (SFP), was extracted from alga Sargassum fusiforme in hot water, was purified by ion exchange chromatography on DEAE-cellulose. The PC, chemical analysis, electrophoresis and IR of SFP indicated that it was a kind of alginate with a mol. wt. of 13,000 and a molar ratio of mannuronic acid to guluronic acid 2.75. Pharmacological tests showed that SFP could prolong the survival duration of mice suffering from ascitic Sarcoma 180 with a rate of life prolongation of 63.44%.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1992년도 제1회 신약개발 연구발표회 초록집
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• pp.24-24
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• 1992

담자균류인 말징버섯 Calvatia craniformis의 성분을 연구하기 위하여 그 균사체를 액내 배양하여 열수 추출한 후 ethanol로 침전시켜 단백 결합 다당체를 분리하였다. 이 성분을 DEAE-cellulose ion exchange, Sepharose CL-4B gel filtration 그리고 Concanavalin A Sepharose 4B affinity chromatography를 실시하여 총 6가지 분획(Fr. A~F)으로 정제하였다. 이들을 각각 10또는 20mg/kg/day의 용량으로 마우스의 복강에 투여하였을때, 육종 180 고형암에 디하여 41.7~74.1%의 종양억제율을 나타내었고 $\beta$-형 다당체인 Fr. F가 종양 억제율이 가장 높았다. 화학 분석에 의해, 이 성분은 4기의 단당 mannose, xylose, galactose, glucose로 구성된 87.2% polysaccharide와 1.9%의 Protein 및 1.3%의 hexosamine 함유하고 있고 이 성분의 분자량은 5.5 $\times$ $10^4$ dalton이었다. 이 성분을 calvatan이라 명명 하였다.

• 한국미생물·생명공학회지
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• 제16권6호
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• pp.438-442
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• 1988

The enzyme produced by Rhizopus japonicus was able to convert selectively ginsenoside-Rb$_1$which is the most abundant ginseng saponin, into ginsenoside-Rd which was known to be superior to ginsenoside-Rb$_1$pharmaceutically. The convertible enzyme was purified homogeneous from wheat bran culture of Rhizopus japonicus by ammonium sulfate fractionation and column chromatography of TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150, Sepharose 2B. Specific activity of the purified enzyme was increased to a bent 96 folds and yield was appeared to be 11% of culture extract. Evidence for homogenity was obtained from polyacrylamide and SDS-polyacrylamide gel electrophoresis. Molecular weight of the enzyme was estimated about 88, 000 daltons by Sephadex G-l50 gel filtration and SDS-polyacrylamide gel electrophoresis, and it did not consist of any subunit.