• 한국식품영양과학회지
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• 제17권3호
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• pp.185-190
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• 1988

아세톤으로 탈지한 대두분으로부터 gel여과법과 이온 교환 크로마토그래피등의 전통적인 단백질 추출 방법을 이용하여 Lipoxygenase 이성효소를 분리하였다. 이온 교환 후의 lipoxygenase-1과 -2는 Crude extract에 비해 각각 19배와 32배 정제 되어졌으나, 상당한 효소활성의 손실을 초래하였으며, lipoxygenase 에 특이적인 염색기술을 이용하여 행한 7% PAGE에서 lipoxygenase-1, -2, 그리고 -3은 각각 0.38, 0.29그리고 0.33의 Rf치를 나타내었다.

• Animal cells and systems
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• 제3권1호
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• pp.47-52
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• 1999

Change of proteins was confirmed during low temperature acclimation of overwintering larva, and some biochemical characteristics of the induced antifreeze protein-1 (AFP-1) were investigated in Protaetia brevitarsis. As the freezing point depression by the action of induced AFPs, a considerable thermal hysteresis was observed in the haemolymph and in partially purified proteins. AFP-1 was purified from the cold acclimation larvae by ammonium sulfate precipitation ion exchange chromatography, gel permeation chromatography, and electroelution. The purified AFP-1 was determined to be a glycoprotein (approximately 320 kDa, pl 5.8) composed of a single type of subunit (80 kDa). The high contents of hydrophilic amino acids (Asp, Glu, Lys, Asn, Gln, Arg, Ser, Thr) were also confirmed, showing similarity with antifreeze proteins from other insects.

• Journal of Microbiology
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• 제38권2호
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• pp.74-79
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• 2000

For the demonstration of a novel riboflavin kinase assay method based on the bacterial bioluminescence, partially purified riboflavin kinase was prepared from bovine liver through ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography. Using bacterial luciferase from Photobacterium phosphoreum and the dithionite reduction method, and easy, safe, and fast assay method was established. The optimal temperature, pH, Km values form riboflavin and ATP of boving liver riboflavin kinase determined with this luminescence method were 35$^{\circ}C$, pH 7, 15.3${\mu}$M and 8.3.${\mu}$M, respectively. The detection limit of FMN produced by riboflavin kinase was in the range of 200 pM to 4${\mu}$M which is comparable to the HPLC-fluorescence detection method, while the detection time for each assay was less than 15 sec compared to the HPLC method which requeires at least 10 min for completion.

• 미생물학회지
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• 제26권1호
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• pp.52-59
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• 1988

A partially purified preparation of L-galactonolactone oxidase which catalyzes the last step of L-ascorbic acid biosynthesis was obtained from Saccharomyces cerevisiae ATCc 26787. The purification procedures included Triton X-100 treatment, protamine sulfate precipitation, ammonium sulfate precipitation, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-150 gel filtration chromatography, and Phenyl-Sepharose CL-4B hydrophobic interaction chromatography. The optimum temperature for the enzyme activity was about $34^{\circ}C$ and the optimum pH was 6.8-7.0. The substrate specificity was confined to L-aldonolactones, L-galactono-1,4-lactone and L-gulono-1,4-lactone. An apparent Km value of 0.294mM with L-galactono-1,4-lactone as a substrate was found. By comparing the substrate specificities of this enzyme with those of isofunctional enzymes of higher plants and animals, it becomes evident that the enzyme of S. cerevisiae ATCC 26787 is rather similar to the L-gulonolactone oxidase of animals than the galactonolactone dehydrogenase of higher plants.

• 한국미생물·생명공학회지
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• 제27권6호
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• pp.446-451
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• 1999

The protease produced by Mucor racemosus f. racemosus PDA 103 from meju was purified by precipitating with 80% saturated ammonium sulfate, CM Sephadex C-50 ion-exchange chromatography, and secondary Sephadex G-100 gel filtration chromatography. The specific activity of the purified enzyme was 60.1unit/mg protein and the purification fold of the enzyme was 83.5. The molecular weight of the enzyme was estimated 33,746Da and the enzyme was elucidated as monomer by LC-MS and SDS-PAGE. The number of amino acids was evaluated about 330 residues.

• 한국미생물·생명공학회지
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• 제23권3호
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• pp.322-328
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• 1995

A thermophilic bacteria showing proteolytic activity against defatted soybean was isolated from soil. It was identified as Bacillus amyloliquefaciens based on its morphological and physiological characteristics. The Bacillus amyloliquefaciens NS 15-4 was cultivated at 50$\circ$C by rotary shaking in a medium containing defatted soybean. An extracellular protease from this strain was purified to homogeneity by ammonium sulfate precipitation, ion exchange, and hydrophobic interaction chromatographies. The molecular weight of the enzyme was estimated to be approximately 30,000 by SDS-PAGE and the N-terminal amino acid sequence of the enzyme was turned out to be AQSVPYGISQIKAPA. The optimum temperature and pH for the enzyme reaction were 60$\circ$C and 11, respectively, and its thermostability was increased by the addition of calcium ion. The enzyme was inactivated by phenylmethylsulfonylfluoride, suggesting it be a serine protease. Comparing with other commercial proteases, the enzyme showed relatively high proteolytic activity against defatted soybean, a water-insoluble protein substrate.

• Preventive Nutrition and Food Science
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• 제3권2호
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• pp.152-156
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• 1998

A Fructan-degrading enzyme was partially purified from onion (Allium cepa)bulbs by a combination of ammonium sufate precipitation, concanavalin-A-Affinity chromatography, and ion-exchange and gel-filtration chromatography. The enzyme hydrolyzed sucrose more effectively than inulin and was identified as a $\beta$- fructofuranosidase (invertase). The optimum pH and temperature were pH 5.5 and 35$^{\circ}C$, respectively. The enzymehydrolyzed sucrose with a Km of 1.2mM . The soluble $\beta$-fructofuranosidase is likely glycoprotein based on its ability to bind the lectin concanavalin-A. The enzyme was heatlabie, with mose activity being lost at 5$0^{\circ}C$ in 1 hr of incubation. The onion $\beta$-fructofuranosidase was partially inhibited by ZnCl2 HgCl2 and CuSo4.

• Nuclear Engineering and Technology
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• 제4권2호
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• pp.83-89
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• 1972

중성자 선속 모니타로써 단 수명 방사성 동위원소인 $^{56}$ Mn을 이용한 새로운 비교법을 사용하여 모나자이트중에 함유된 개개 희토류원소의 함량을 중성자 방사화법으로 정량하였다. 전회토류원소를 군으로 분리하고 난 다음 중성자 조사를 하였으며 개개 희토류 원소는 양이온 교환수지통을 통하여 알파 하이드록씨 부탈산으로 구배 용출시킴으로서 분리하였다. 루데슘으로 부터 란타늄까지 14개의 희토류원소를 정량하였다.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.146.1-146.1
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• 2003

A major protein was isolated from ginseng root (Panax ginseng C.A. Meyer) using a combination of ammonium sulfate fractionation, gel filtration chromatography, ion-exchange FPLC. Electrophoretic and gel permeation chromatographic studies revealed that the major protein, GMP, is composed of two subunits of approximately 28 kDa. In this, investigated the ability of GMP to inhibit the oxidation of low-density lipoprotein (LDL). GMP inhibited $Cu^{2+}$ (5$\mu$M)-promoted oxidation of LDL (125$\mu$g protein/mL) in a dose-dependent mamer (0~5 $\mu$M), with a maximal inhibitor at GMP/copper ratio of 1:10 and an $IC_{50}$ value of 0.2 $\mu$M, as determined by measurement TBARS. (omitted)

• 멤브레인
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• 제32권5호
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• pp.348-356
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• 2022

바이오산업의 발전으로 의약품, 식품 등의 생산 과정의 분리/정제 공정에 사용되어 왔던 기존의 컬럼 크로마토그래피를 대체하여 더 높은 처리효율을 갖는 막 크로마토그래피가 부상하고 있다. 본 연구에서는 서로 다른 기공 크기의 두 가지 상용 셀룰로오스 아세테이트(Cellulose acetate, CA) 분리막을 탈아세틸화 과정을 통해, 리간드의 개질이 용이한 다공성 재생 셀룰로오스 지지체를(Regenerated cellulose, RC) 제조하였다. 음이온 교환능을 부여하고자 grafting을 수행하였으며, 구체적으로는 UV 중합법을 통해 4차 암모늄을 포함하는 음이온 교환 리간드(MAPTAC)를 부착하여 음이온 교환용 흡착막을 제조하였다. 단백질 흡착 용량은 정적 흡착 용량(Static binding capacity, SBC)시험을 통해 총 단백질 흡착 용량을 측정했고, 동적 흡착 용량(Dynamic binding capacity, DBC)을 측정하여 상용막과 비교 평가하였다. 성능 평가 결과 단백질 흡착량은 넓은 표면적에 의해 리간드 밀도가 높은, 기공 크기가 작은 순서로 높게 측정되었고, 상용 CA분리막을 탈아세틸화하고 리간드를 부착시킨 분리막(RC 0.8 + MAPTAC 43.69 mg/ml, RC 3.0 + MAPTAC 36.33 mg/ml)이 상용 막 크로마토그래피 제품(28.38 mg/ml) 대비 높은 흡착 용량을 보였다.