• 한국미생물·생명공학회지
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• 제18권4호
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• pp.344-351
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• 1990

계명활성제 내성이 있으면서 호알카리성인 protease를 생산하는 미생물을 토양에서 분리하였다. 분리된 미생물을 형태적, 생리학적, 화학분류학적 및 5S RNA 분석으로 동정한 결과 Bacillus sp.인 것으로 판명되었다. 호알카리성 protease는 황산암모늄 분획, DEAE-Cellulose, CM-Cellulose, Sephadex G-100 column chromatogrphy로 분리, 정제하였다. 정제된 호알카리성 protease는 casein에 대하여 pH6.0에서 11.0 사이에서 안정성을 나타내었다. 분리된 효소의 작용 최적 온도는 $55^{\circ}C$이었다. 이 효소는 diisopropyl fluorophosphate(DFP)로 완전히 불활성화되는 것으로 보아 serine protease로 추정되며 계면활성제의 존재하에서도 안정하였다.

• Journal of Yeungnam Medical Science
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• 제13권1호
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• pp.46-58
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• 1996

세포막 정보전달 과정에 참여하는 효소로 inositol triphosphate($InsP_3$)를 분해하는 $InsP_3$ kinase를 소의 뇌 조직으로 부터 새로운 정제방법을 개발하고 isozyme들의 특성을 관찰하여 다음과 같은 결과를 얻었다. 분쇄한 소의 뇌조직을 PEG로 단백침전하고 DEAE cellulose chromatography 하여 $InsP_3$ kinase I, II의 두 isozyme을 얻었으며, 각각의 isozyme 분획을 Matrix green gel 및 calmodulin-Affigel 15 column으로 chromatography하였다. Phenyl-TSK HPLC하여 정제하였으며 I은 3,103배, II는 2,310 배의 정제를 나타내었다. 정제 단계에서 specific activity를 비교해 볼 때 Matrix green gel chromatography가 DEAE cellulose에서 보다 I이 30배, II가 약 2.3배로 나타났고 정제배수도 I이 17.2%에서 62.1%로 II가 16.6%에서 38%로 나왔으나 calmodulin-Affigel 15과 비교시는 큰 차이가 없었다. 그러므로 Matrix green gel이 정제에서 매우 중요한 단계라 할 수 있다. 효소의 분자량을 알기 위하여 DEAE HPLC로 두 개의 isozyme을 분리하여 전기영동한 결과 I은 환자량 145,000, 85,500 및 69,500이 3개의 단백질을 얻었고 II는 분자량 79,000 및 57,000의 2개의 단백질을 얻었다.

• 미생물학회지
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• 제14권2호
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• pp.65-74
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• 1976

Atternaria sp. was isolated from soil and crude cellulases were prepared from wheat bran culture of the fungus. The activities of the crude enzyme were studied on five different subvstrates and some phsical properties were also examined, crude enzymes were purified by column chromatography on DEAE Sephadex and Sephadex, Isozymes were separated some of which were active specifically on DEAE cellulose and some were primarily active on cellulose and CM-cellulose. The optimal points of pH and temperature for the crude enzyme were varied depending on the substrates ; On cellulose they were at pH 6.0 and 40.deg.C, on CM-cellulose at pH's 4.0 and 6.0 and 60.deg.C, and on DEAW-cellulose at pH 5.0 and 50.deg.C. Two active fractions, F-1 and F-II on Na-CMC was used as substrate the Km values of crude enzyme, F-I and F-II were calculated to be $4{\times}10^{-5}$ , 1.1 * 10$^{-4}$ , and $1.25{\times}10^{-4}mN$ resepctively. The Ki value of $Cu^{++}$ for crude enzyme was$4{\times}^{-4}mN$ , while that of $Nm^{++}$ while in the same concentration of $Mn^{++}$ it reached to 91%. Some 57% activity of F-1 was inhibited in s mN $Cu^{++}$, whereas it was inhibited as much as 81% in the same concentration above the concentration of 0.3 mM with tis activity reaching up to 137% in 2 mM. On the other hand the F-11 was inhibited by the presence of M $n^{++}$ and some 67% activity was inhibited at 2mM.

• Parasites, Hosts and Diseases
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• 제21권1호
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• pp.41-48
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• 1983

The activity and distribution of aspartate aminotransferase (EC 2.6. 1. 1) and alanine aminotransferase (EC 2.6.1.2) in adult Fascicle hepatica have been studied. Fasciola hepatica was fractionated by differential centrifugation into nuclear, mitochondrial and cytosolic fractions. The activity of GOT and GPT was measured by the method of Reitman and Frankel. Isozyme patterns of those enzyme were also examined by DEAE-cellulose column chromatography. The results obtained were as follows; 1. The activity of aspartate and alanine aminotransferase was about 0.55 unit and 0.92 unit per 1g of Fascicle hepatica, respectively. 2. The activity of those enzymes was relatively low compared with those in mammalian tissues. 3. The distribution of aspartate aminotransferase in the subcellular organelles showed that 71% of the activity was in cytosolic, 24% in mitochondrial and 5% was in nuclear fraction. 4. About 22% of the total alanine aminotransferase activity was found in the mitochondrial fratstion, about 66% in the cytosolic fraction. 5. Aspartate aminotransferase from cytosolic fraction was separated into two types of isozymes, whereas alanine aminotransferase from cytosolic fraction gave only one active peak on DEAE-cellulose column chromatography.

• 한국동물학회지
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• 제31권4호
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• pp.300-308
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• 1988

Human placental alkaline phosphatase (PLAP), one of the oncofetal antigen was purified from placentas through the procedures including butanol extraction, concanavalin A-Sephar-ose, DEAE-cellulose and Sephadex G-200 gel chromatography. Monoclonal antibodies (fibs) against human PMP were produced by hybridizing SP 210-Ag 14 mouse myeloma cells with spleen ceils of Balblc mice immunized with PLAP. Six stable monoclones uvere obtained by cloning tuvice in serial dilutions, and the monoclonal speclfidty of these MAbs was confirmed by biochemical and immunonogical criteria. Tumor marker의 하나인 태반형 alkaline phosphatase(PLAP)에 대한 단일 클론항체의 생산과 분석을 위하여, 태반조직을 재료로 butanol 추출법 및 concanavaline A-Sepharose, DEAE-cellulose, Sephadex G-200 gel 크로마토그라피법에 의하여 PLAP를 순수 분리하였다. 이를 항원으로 하여 하이브리도마 방법에 의해 항-PLAP 단일 클론항체를 생산 분비하는 안정된 6클론세포를 얻었으며 생화학적 및 면역학적 분석방법으로 이들의 단일 클론성을 확인하였다.

• Applied Biological Chemistry
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• 제13권3호
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• pp.223-229
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• 1970

(1) 고온성사상균인 Myriococcum 속의 알칼리성 protense의 생성과 조효소의 성질을 검토하였다. (2) 조호소의 최적 pH는 9.0이고, 최적 온도는 $55^{\circ}C$였다. 그리고 안정 pH 범위는 6.0에서 11.0이었다. (3) 조효소용액을 DEAE-Cellulose column Chromatography로 2개의 다른 활성부분으로 나누어졌다.

• 한국미생물·생명공학회지
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• 제15권6호
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• pp.420-424
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• 1987

Saccharomycopsis lipolytica ATCC 44601 과 MX9-11RX8 온도감수성 변이균주의 isocitrate lyase는 조추출액을 ammonium sulfate 분획, Toyo Peal HW-55F gel filtration, DEAE-Cellulose ion exchange chromatography 등의 방법에 의하여 각각 54배, 87배 분리 정제되었다. 정제효소의 subunit 분자량은 59,000이고 Sephadex G-200 gel filtration에 의한 native enzyme 은 230,000이므로 이 효모의 isocitrate lyase는 같거나 비슷한 subunit 4개로 구성된 tetramer이며, 최적 pH는 6.9이었다.

• 한국동물학회지
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• 제29권4호
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• pp.283-293
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• 1986

사람의 alpha-fetoprotein(AFP)에 대한 모노클론 항체의 생산 및 분석을 위하여, 태아조직을 재료로 추출법, DEAE-cellulose 및 concanavalin A-Sepharose, Cibacron blue F3GA-agarose, immunoadsorbent column등의 흡착크로마토그라피법에 의해 AFP를 분리하였다. 총 534g의 태아조직에서 AFP의 量은 8.76 mg으로서 순수분리되었음을 확인하였다.

• 한국축산식품학회:학술대회논문집
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• 한국축산식품학회 1995년도 추계심포지움 및 학술발표회
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• pp.104-104
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• 1995

• Mycobiology
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• 제37권2호
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• pp.121-127
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• 2009

Purification and characterization of intracellular cellulase produced by A. oryzae ITCC-4857.01 are reported. The enzyme was purified by ion-exchange chromatography using DEAE-cellulose followed by Gel filtration. The purification achieved was 41 fold from the crude extract with yield of 27%. The purified enzyme showed single band on poly acrylamide gel. The molecular weight as determined by SDS-PAGE and gel filtration was 38 KDa and 38.6 KDa respectively and contained only one subunit. The enzyme is glycoprotien as nature and contained 0.67% neutral sugar. The apparent Km value of the enzyme against cellulose was 0.83%. The enzyme showed the highest relative ativities on CMC followed by avicel, salicin and filter paper. The optimum pH of activity was 5.5 and very slight activity was observed at or above pH 7.5 as well as bellow pH 3.5. The optimum tempreture of the activity was $45^{\circ}C$ and the highest activity was exhibited in 35 to $45^{\circ}C$. The enzyme lost their activities almost completely (95${\sim}$100%) at $80^{\circ}C$ or above and as well as bellow $25^{\circ}C$.