• Applied Biological Chemistry
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• 제30권1호
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• pp.88-94
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• 1987

인삼 성분중 생리적 활성을 가지고 있는 oligopeptide를 검출하고 분리하기 위하여 물 추출액을 ultra-filtration 한 후 gel filtration, ion-exchange chromatography 및 thin layer chromatography 등을 행하였다. 인삼 물 추출액으로부터 고분자 물질을 제거한 ultra-filtrate는 epinephrine에 의해 유도된 fat cell의 lipolysis를 저해하는 antilipolytic activity를 보유하고 있었으며 Sephadex G-25 gel filtration에 의해 3개 fraction으로 나뉘어졌다. 이중 첫번째 fraction(S-FI)만이 peptide 성분을 함유하고 있었으며 saponin 및 당도 검출되었다. S-FI fraction은 $Dowex\;50{\times}2\;ion-exchange\;chromatography$에 의하여 6개의 $fraction(P-F1{\sim}P-F6)$으로 분리되었고 TLC검정 결과 P-F2 fraction이 peptide fraction으로 antilipolytic를 보유하고 있었으며 TLC로 분리한 6개 spot는 각각 가수 분해한 전후의 TLC pattern을 비교해 본 결과 모두 oligopeptide임이 밝혀졌다. S-FI fraction에 존재한 saponine과 당은 P-F1 fraction 에서 모두 용출되었다.

• 한국응용곤충학회지
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• 제37권1호
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• pp.59-64
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• 1998

꿀벌부채명나방 종령유충의 whole body에서 gel filtration 방법으로 유약호르몬 결합 단백질을 분리, 정제하였다. 분리된 단백질은 column chromatography법과 전기영동법에 의해 등가성을 확인하였다. 이 결합단백질은 전기 영동법에 의해 32K, gel filtration 에 의해 28K의 상대적 분자량을 나타냈다. 또한, JH III에 대한 해리도는 3.9$\times$${10}^{-7}$M로 확인되었다.

• 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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• 제34권2호
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• pp.196-202
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• 1991

매미나방 종령유층 혈림프내에 존재하는 JHBP을 Dextran Coated Charcoal (DCC)binding assay와 gel filtration에 의해서 확인하였고, JHBP의 pI값은 5.3으로 밝혀졌다. JHBP의 정저는 혈림프단백질을 먼저 PEG로 침전시킨 후 ion exchange chromatography와 gel filtration 방법을 통하여 행하였다. 정체된 fraction의 JH에 대한 binding activity는 [3H] JH-III의 radioactivity 측정과 DCC binding assay를 통해 확인하였고, 정체된 단백질의 순수도는 각 정체단계에 따라 전지영동을 하여 확인하였다.

• KSBB Journal
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• 제17권4호
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• pp.365-369
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• 2002

수용성 형태로 존재하는 융합 페리틴을 정제함에 있어서 실리카 분말을 이용한 전처리 공정은 전체 정제 공정효율 증가에 기여하였다. 전처리 공정을 통해 정제된 융합 페리틴의 순도를 높이기 위해 젤 여과 크로마토그래피를 통해 보다 정제된 응합 페리틴을 얻을 수 있었다. 이렇게 정제된 융합 페리틴의 철분결합능력을 분석해본결과 320 moles $F_{H}+F_{L}$ mole로 활성이 우수함을 알 수 있었으며, 분자량 분석을 통해 융합페리틴(40 k dalton)은 trimer와 monomer형태로 존재함을 확인할 수 있었다

• 한국미생물·생명공학회지
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• 제17권6호
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• pp.587-592
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• 1989

토양으로부터 분리한 호알카리성 Bacillus sp. YS-309의 조효소액을 조제하고 제핵산, ammonium sulfate 침전, DEAE-cellulose column chromatography, Sephacryl S-200 gel-filtration, DEAE-Sephadex A-50 chromatography 등을 단계적으로 수행하여 6.9배 정제된 순도 98%의 정제효소를 얻었으며, 활성염색을 실시하여 정제한 효소단백질이 $\beta$-galactosidase임을 확인하였다. 정제효소의 분자량은 205,000으로 monomer의 분자량이 56,000인 동일크기의 tetramer로 구성되어 있다고 판단되었다.

• BMB Reports
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• 제28권3호
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• pp.197-203
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• 1995

Famesyl protein transferase involved in the first step of post-translational modification of $p21^{ras}$ proteins transfers the famesyl moiety from famesyl pyrophosphate to a cysteine residue in $p21^{ras}$ proteins. The enzyme was first purified 30,000-fold from bovine testis by use of 30~50% ammonium sulfate fractionation, DEAE-Sephacel ion exchange chromatography, Sephacryl S-300 gel filtration chromatography, Sephacryl S-200 gel filtration chromatography, and hexapeptide (Lys-Lys-Cys-Val-Ile-Met) affinity chromatography. The molecular weight of the purified enzyme was estimated to be ~100 kDa by gel filtration and SDS-polyacrylamide gels showed two closely spaced bands of ~50 kDa protein. These indicate that the enzyme consists of two nonidentical subunits, a and 13, which have slightly different molecular weights. The enzyme was inhibited by hexapeptide (Lys-Lys-Cys-Val-Ile-Met), which acted as an alternative substrate that competed for famesylation. Kinetic analysis by measuring initial velocities showed that famesyl protein transferase is a very slow enzyme. EDTA-treated famesyl protein transferase showed little activity with $Mg^{2+}$ or $Zn^{2+}$ alone, but required both $Mg^{2+}$ and $Zn^{2+}$ for the catalytic activity.

• Fisheries and Aquatic Sciences
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• 제17권2호
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• pp.181-187
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• 2014

This study was performed to identify the optimum fractionation method and conditions to obtain exopeptidase-active fractions from octopus hepatopancreas (HP) crude extracts (CEs) using four techniques: solid ammonium sulfate fractionation, polyethylene glycol (PEG) fractionation, anion exchange chromatography, and gel filtration chromatography. The fractions with the highest total activity toward L-leucine-p-nitroanilide (Leu-pNA) were fraction IV from the ammonium sulfate and PEG fractionation, and fraction II in ion exchange and gel filtration chromatography. The total exoprotease activity of these fractions was highest in fraction IV (4,050.20 U) of ammonium sulfate fractionation, followed by fraction II (3,600.28 U) from gel filtration chromatography, fraction IV (2,861.30 U) from PEG fractionation, and fraction II (2,576.28 U) from ion exchange chromatography. These results suggest that ammonium sulfate fractionation using 60-80% ammonium sulfate was the most efficient method for separating the exoprotease active fractions from CEs of octopus HP.

• 한국수산과학회지
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• 제46권4호
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• pp.351-358
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• 2013

A protease inhibitor from fish eggs was fractionated using chromatographic methods. The fractionation efficiency was evaluated in terms of specific inhibitory activity (SIA, U/mg), purity (fold), total inhibitory activity (TIA, U), and recovery (%). The protease inhibitor (PI) from egg extracts of skipjack tuna (ST Katsuwonus pelamis), yellowfin tuna (YT Thunnus albacares) and Alaska pollock (AP Theragra chalcogramma) was fractionated using Sephadex G-50 gel filtration and DEAE-Sepharose CL-6B anion exchange chromatography based on protein size exclusion and net charge, respectively. Fractions exhibiting strong inhibitory activity were contained in the 30-50 kDa fraction on gel filtration and in the range of 0.4-0.7 M NaCl gradient fraction on anion exchange chromatography. The respective TIA and percent recovery of the fraction obtained with gel filtration toward trypsin and $N{\alpha}$-benzoyl-L-arginine-p-nitroanilide (BAPNA) were 2,758.7 U and 29.6% for ST, 1,005.5 U and 25.6% for YT, and 1,267.5 U and 26.0% for AP. Gel filtration chromatography was more effective at fractionating PI than using ion exchange chromatography. These results suggest that fish eggs act as serine protease inhibitors and might be useful for protease inhibition in foodstuffs.

• BMB Reports
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• 제35권6호
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• pp.576-582
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• 2002

Collagenase from the internal organs of a mackerel was purified using acetone precipitation, ion-exchange chromatography on a DEAE-Sephadex A-50, gel filtration chromatography on a Sephadex G-100, ion-exchange chromatography on DEAE-Sephacel, and gel filtration chromatography on a Sephadex G-75 column. The molecular mass of the purified enzyme was estimated to be 14.8 kDa by gel filtration and SDS-PAGE. The purification and yield were 39.5-fold and 0.1% when compared to those in the starting-crude extract. The optimum pH and temperature for the enzyme activity were around pH 7.5 and $55^{\circ}C$, respectively. The $K_m$ and $V_{max}$ of the enzyme for collagen Type I were approximately 1.1 mM and 2,343 U, respectively. The purified enzyme was strongly inhibited by $Hg^{2+}$, $Zn^{2+}$, PMSF, TLCK, and the soybean-trypsin inhibitor.