• 제목/요약/키워드: sulfate fractions

검색결과 132건 처리시간 0.025초

Comparison of the Exopeptidase Activity of Fractions from Crude Extracts of Octopus Octopus vulgaris Cuvier Hepatopancreas Using Different Fractionation Methods

  • Kim, Min Ji;Kim, Hyeon Jeong;Kim, Ki Hyun;Heu, Min Soo;Kim, Jin-Soo
    • Fisheries and Aquatic Sciences
    • /
    • 제17권2호
    • /
    • pp.181-187
    • /
    • 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.

인삼(人蔘) Saponin이 혈장(血漿) Lipoproteins 및 Lecithin Dispersion과 Dextran Sulfate의 상호작용(相互作用)에 미치는 영향(影響) (Studies on the Effect of Panax ginseng on the Interactions of Human Plasma Lipoproteins and Lecithin Dispersions with Dextran Sulfate)

  • 김영중;전미희
    • 생약학회지
    • /
    • 제15권4호
    • /
    • pp.206-212
    • /
    • 1984
  • The effect of saponin fractions of Panax ginseng root on the interactions of human plasma lipoproteins and lecithin dispersions with dextran sulfate were studied in order to examine the effect of Panax ginseng on the lipid accumulation in the aorta. The total saponin fraction and protopanaxadiol glycosides of Panax ginseng root seemed to slightly enhance the interaction of low density lipoproteins with dextran sulfate in the absence of divalent metal ions. Protopanaxatriol glycosides remarkably inhibited the interaction of low density lipoproteins with dextran sulfate. However, all of these three saponin fractions of Panax ginseng root showed the tendency of inhibition to the interaction of high density lipoproteins with dextran sulfate in the presence of divalent metal ions by the order of protopanxatriol glycosides, protopanaxadiol glycosides and total saponin. Three saponin fractions of Panax ginseng exerted almost same tendency to the interaction of lecithin dispersions with dextran sulfate in the presence of divalent metal ions as the interaction of low density lipoproteins with dextran sulfate absence of divalent metal ions.

  • PDF

혈청단백질분획(血淸蛋白質分劃)에 대한 연구(硏究) (I, II) (Studies on Bovine Serum Protein Fractions (I, II))

  • 임봉호
    • 대한수의학회지
    • /
    • 제5권1호
    • /
    • pp.1-16
    • /
    • 1965
  • I. A Comparison of Sodium Sulfate Precipitation and Zone(Paper, Agar) Electrophoresis; Many kinds of techniques have been used for fractionating serum proteins. In the present study, using bovine serum, the fractions obtained with sodium sulfate were compared with those determined by zone electrophoresis. 1. Fibrinogen was precipitated with 4 to 10 percent of sodium sulfate. 2. ${\gamma}$-globulin required 10 to 16 percent of the salt for precipitation. 3. ${\beta}$-globulin began to precipitate at 12 percent sodium sulfate, and completed precipitation at approximately 26 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. 4. ${\alpha}$-globulin completed precipitation at 13 to 28 percent sodium sulfate in paper electrophoresis and at 22 percent in agar electrophoresis. 5. Albumin began to precipitate at 14 percent of the salt, and was free from the mixture of globulins approximately at 28 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. The results of comparing fractions by the two methods were as follows: 1. Euglobulin (15%) was equal to the sum of the most ${\gamma}$-globulin and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. 2. Pseudoglobulin I (15-17.5%) corresponded to the most ${\alpha}$-, ${\beta}$-globulins and a small quantity of albumin. 3. Pseudoglobulin II(18-22%) was a mixture of the ${\alpha}$-, ${\beta}$-globulins and albumin fraction. 4. Albumin (above 22%) contained the most albumin fraction separated by zone electrophoresis and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. As mentioned above the fractions obtained with sodium sulfate were a mixture of the various proportion of the fractions determined by zone electrophoresis. The solubility of serum fractions to sodium sulfate coincided with the mobility of those by zone electrophoresis. (By percent of sodium sulfate we mean gram of sodium sulfate contained in $100m{\ell}$ of solution). II. Immunological Studies on Serum Protein Fractions with Sodium Sulfate; In the previous report the fractions of bovine serum protein with sodium sulfate compared with those obtained by zone electrophoresis, and the findings were that the former contained various proportion components of the latter. In this study the author studied whether or not the fractions with sodium sulfate are simple component antigenically by immunoelectrophoresis and micro double diffusion test (Immuno-precipitation), using rabbit antiserum to bovine serum. In immunoelectrophoresis, normal bovine serum developed with rabbit antibovine serum showed about ten distinct precipitin arcs. The distribution of these arcs was as follows: 1 albumin, 2 ${\alpha}_1$-, 3 ${\alpha}_2$-, 2 ${\beta}_1$-, ${\beta}_2$-, and 1 ${\gamma}$-globulin (Fig. 7, 9). In micro double diffusion test, five to six precipitation bands could be seen between antigens and antibody, the order of the precipitation bands location is albumin, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-globulin from the side of antiserum well (Fig.19). Frequently the ${\alpha}$-, and ${\beta}$-precipitation bands were separated into two or three precipitation bands, which indicated that these globuline are not a pure component antigenically as shown in immuno-electrophoresis. In both Immunological methods, the two ${\alpha}$-, ${\beta}$-precipitin arcs and bands appeared clear and strong, indicating that the two globulins reacted as strong antigens. The precipitate reaction of ${\gamma}$-globulin was shown at 12 to 16 percent sodium sulfate; ${\beta}$-globulin at 12 to 20 percent; ${\alpha}$-globulin at 12 to 22 percent (immuno-electrophoresis), at 12 to 26 percent (Diffusion); and albumin at above 22 percent. Antigenically euglobulin contained ${\gamma}$-, ${\beta}$-, and ${\alpha}$-globulins, Pseudoglobulin I and Pseudoglobulin II were composed of ${\alpha}$-, and ${\beta}$-globulins, and albumin was a mixture of ${\alpha}$-globulin and albumin determined by zone electrophoresis. The results indicated that the fractions of serum protein obtained by either method were constituents of various proteins antigenically except ${\gamma}$-globulin and albumin by Zone electrophoresis.

  • PDF

培養 鷄胚 筋原細胞의 融合에 미치는 鷄胚 抽出液 分劃의 영향 (The Effects of Fractions of Chick Embryo Extract on the Fusion of Cultured Chick Embryonic Myoblasts)

  • Ha, Doo-Bong;Lee, Chung-Choo;Park, Young-Chul;Lim, Woon-Ki;Yoo, Byoung-Je
    • 한국동물학회지
    • /
    • 제28권3호
    • /
    • pp.179-193
    • /
    • 1985
  • 鷄胚의 筋原細胞의 증식과 融合에 미치는 계배 抽出液의 영향을 조사하였다. 1. 배양액내의 계배 추출액의 농도가 높을수록 근원세포의 증식은 촉진되었으며, 반면에 근원세포 융합은 지연되었다. 2. 계배 추출액의 단백질을 Sephadex G-75로 分劃하고, 각 분획을 근원세포의 배양액에 첨가한 결과 分子量 40,000과 22,000 dalton 사이의 분획이 근원세포의 증식과 융합을 촉진시켰다. 3. 계배 추출액의 단백질을 ammonium sulfate로 분획시켜 각 분획을 근원세포의 배양액에 첨가한 결과 70% 이상 포화 용액에서 침전하는 분획이 근원세포의 증식과 융합을 현저히 증가시켰다. 이 유효 분획을 Sephadex G-75로 재차 분획하여 각 분획의 효과를 조사한 결과 근원세포의 증식과 융합을 촉진시키는 분획이 계배 추출액을 Sephadex G-75로 분획하여 얻은 유효 분획과 거의 동일한 효과를 나타내었다.

  • PDF

The Bioactivity of Natural Product in the Ovariectomized Rat

  • Ha, Bae-Jin
    • Journal of Life Science
    • /
    • 제11권1호
    • /
    • pp.47-51
    • /
    • 2001
  • To investigate the deaging effects of introperitoneally injected Chondroitin Sulfate (CS) on various enzyme activity (AST, ALT, MDA (Malon dialdehyde), SOD (Superoxide dismutase), GPx (Glutathione peroxidases) and histophathology of liver tissue, ovariectomized rats were used. The antioxidative effects of chondroitin sulfate (100 mg/kg and 200 mg/kg body weight) were investigated at the antioxidative enzyme activities of liver homogenate fractions (liver total homogenate, mitochondrial, and microsomal fractions) and sera. In addition, the rat liver was histologically examined. Intraperitoneally injected CS, depend on dosage, indicated a protective effect against ovariectomy-inducted aging. Moreover, inflammation and cirrhosis in liver tissue of CS treated group were significantly decreased. Based on these results, intraperitoneally injected CS is a useful material to delay aging.

  • PDF

Fractionation and Enzymatic Characterization of Endoprotease and Exopeptidase from Crude Extracts of Cuttlefish Sepia officinalis Hepatopancreas

  • Kim, Min Ji;Kim, Hyeon Jeong;Kim, Ki Hyun;Heu, Min Soo;Lee, Jung Suck;Kim, Jin-Soo
    • Fisheries and Aquatic Sciences
    • /
    • 제15권4호
    • /
    • pp.283-291
    • /
    • 2012
  • This study examines the optimal fractionation method and conditions for the isolation of endoprotease- and exopeptidase-active fractions from crude extracts of cuttlefish hepatopancreas (HP) using four fractionation methods: ammonium sulfate fractionation (ASF), polyethylene glycol fractionation (PGF), ion exchange chromatography (IEC), and gel filtration chromatography (GFC). Total endoprotease activity highest in the fraction II (concentrate of fractions 34-42; 842.60 U) of GFC, followed by fraction III (40-60% ammonium sulfate fraction; 670.25 U) of ASF, fraction I (concentrate of fractions 8-12; 436.89 U) of IEC, and fraction II (10-20% polyethylene glycol; 307.31 U) of PGF. Total exopeptidase activity of these fractions was highest in fraction II (2,704.70 U) of GFC, fraction III (2,110.50 U) of ASF, fraction III (1,605.60 U) of PGF, and fraction II (concentrate of fractions 38-44; 1,196.22 U) of IEC. These results showed that fraction II of GFC had the highest activity toward both exopeptidase and endoprotease, with exopeptidase activity being 3.21 times higher than of endoprotease. These results suggest cuttlefish HP could be used as a potential source for the extraction of exopeptidase, an enzyme capable of catalyzing the cleavage of N- and C-terminal amino acids in polypeptides, Like endoprotease, the most efficient method for separating exopeptide-active fractions was GFC.

다양한 유기성 폐자원을 적용한 바이오 수소 생성 연구 (Feasibility of batch anaerobic bio-hydrogen production from different organic wastes)

  • 황재훈;최정아;;전병훈
    • 신재생에너지
    • /
    • 제5권4호
    • /
    • pp.80-85
    • /
    • 2009
  • The effects of various organic wastes on anaerobic fermentative hydrogen production were studied using enriched mixed microflora in batch tests. Rotten fruit, corn powder and organic wastewater enriched with sulfate (up to 1,000 mg/L) were used for experiments. Maximum hydrogen production (547.1 mL) was observed from rotten apple with initial substrate concentration of 132.2 g COD/L. The experimental result on sulfate enriched organic wastewater indicated that hydrogen production is not adversely influenced by relatively high sulfate concentration. Residual sulfate content remained at 96-98 % after 75 hours of reaction, which showed that no major sulfate reduction was occurred at pH 5.3-5.5 in the reactor. The volatile fatty acid (VFA) fractions produced during the reaction was in the order of butyrate > acetate > propionate in all experiments. The results of this study would be useful for controlling the conditions on fermentative hydrogen production using different feedstocks.

  • PDF

폭약 TNT 분해세균 Stenotrophomonas sp. OK-5에서 분리된 NAD(P)H-nitroreductase의 정제 및 특성 연구 (Characterization of NAD(P)H-nitroreductase Purified from the TNT-degrading Bacterium, Stenotrophomonas sp. OK-5)

  • 호은미;천재우;강형일;오계헌
    • 미생물학회지
    • /
    • 제39권4호
    • /
    • pp.223-229
    • /
    • 2003
  • 2,4,6-Trinitrotoluene (TNT)을 분해할 수 있는 Stenotrophomonas sp. OK-5에서 분리한 NAD(P)H-nitroreductase의 특성을 조사하였다. 먼저 ammonium sulfate precipitation, DEAE-sepharose, 그리고 Q-sepharose 등의 일련의 정제 과정을 통하여 NAD(P)H-nitroreductase을 분리정제하였다. 분취기로부터 얻어진 시료로부터 NAD(P)H-nitroreductase의 효소활성을 가지는 3개의 다른 fractions (I, II 및 III)이 탐침되었다. NAD(P)H-nitroreductase의 fractions I, II, 그리고 III의 비활성(specific activity)은 각각 5.06 unit/mg, 4.95 unit/mg, and 4.86 unit/mg이었으며, crude extract와 비교하여 각각 10.5배, 9.8배, 8.9배 이상 농축되었다. 이 실험에서 이들 3개의 fractions 가운데, fraction I이 가장 높은 비활성을 나타내었다. NAD(P)H-nitroreductase (fractions I, II 및 III)의 효소활성에 영향을 미치는 몇 가지 요인을 조사하였다. 모든 NAD(P)H-nitroreductase (fractions I, II 및 III)의 최적 온도는 30$^{\circ}C$이었으며, 최적 pH는 약 7.5이었다. 4Ag^+, Cu_2^+, Hg_2^+$ 등의 금속이온은 약 80%의 효소활성을 저해하였으나, $Mn_2^+ 이나 Ca_2^+$의 첨가 시에는 약 30~40% 정도의 활성이 감소되었다. 그러나 $Fe_3^+$은 이들 효소의 활성을 증진시켰다. SDS-PAGE에 의해 측정된 NAD(P)H-nitroreductase의 fractions I, II 및 III의 분자량은 모두 약 27 kDa임이 확인되었다.

원양산 오징어(Illex argentinus) 내장으로부터 Endoprotease의 분획 (Fractionation of Endoprotease from Viscera of the Argentina Shortfin Squid Illex argentinus)

  • 김혜숙;김진수;허민수
    • 한국수산과학회지
    • /
    • 제41권3호
    • /
    • pp.176-181
    • /
    • 2008
  • To evaluate the effective use of endoprotease from squid viscera as a food processing aid, various methods of fractionating endoprotease from viscera of the Argentina shortfin squid (Illex argentinus) were evaluated. The endoprotease-positive fractions of each fractionation were fraction II (30-40%, w/w) with cold acetone, fraction IV (50-60% saturation) with ammonium sulfate, fraction UF with anion exchange chromatography, and fraction II (15-24 kDa) with gel filtration. The specific activities (approximately 25 U/mg) of the fractions using ammonium sulfate and gel filtration were higher than the others. Total azocaseinolytic activity and recovery of the positive fraction using gel filtration were 806.95U and 37.82%, respectively, and were the highest among the positive fractions. Based on the results, gel filtration was the most efficient method for fractionating endoprotease from the viscera of Illex argentinus.

폭약 TNT를 분해하는 세균인 Pseudomonas SP. HK-6에서 분리정제된 Nitroreductase의 특성연구 (Characterization of Nitroreductase Purified from TNT-degrading Bacterium, Pseudomonas sp. HK-6.)

  • 호은미;강형일;오계헌
    • 한국미생물·생명공학회지
    • /
    • 제32권3호
    • /
    • pp.230-237
    • /
    • 2004
  • 균주 Pseudomonas sp. HK-6로부터 2,4,6-trinitrotoluene (TNT)의 대사 과정에서 유도되는 nitroreductase (NTR)를 분리 및 정제하여 다양한 특성조사를 실시하였다. NTR은 균주 HK-6의 세포추출물로부터 ammonium sulfate 침전, DEAE-sepharose, 그리고 Q-sepharose chromatography의 일련의 과정을 통하여 분리되었고, NTR의 활성을 가지는 세개의 다른 fractions를 확인하였다. 균주 HK-6의 NTR fractions I, II, 그리고 III의 비 활성은 각각 4.85 unit/mg, 5.47 unit/mg, 5.01 unit/mg으로 측정되었으며, 세포추출물에 비해 각각 9.0배, 10.1배, 9.3배 농축된 것으로 나타났다. SDS-PAGE에서 측정된 균주 HK-6의 NTR fractions I, II 그리고 III의 분자량은 모두 약 27 kDa으로 확인되었다. 정제된 NTR의 활성에 온도, pH, 금속 이온, 억제 물질의 효과와 기질 특이성 등에 대한 물리화학적 특성 조사를 실시하였다. 균주 HK-6의 NTR fractions I, II, 그리고 III의 적정온도는 $25~35^{\circ}C$ 확인되었고, 모두 $30^{\circ}C$에서 최대 활성을 나타내었으며, 이들 효소 활성의 적정 pH는 7.0~8.0이었고, 최적 pH는 7.5로 확인되었다. TNT에 대한 HK-6의 NTR fractions I, II, 그리고 III의 활성은 금속 이온 $Ag^{+}$ , $Cu^{2+}$ , 그리고 $Hg^{V}$ 에 의해서 약 70%이상 저해되었으며, $Mn^{2+}$ 또는 $Ca^{2+}$ 에 의해서 약 20~50%로 활성이 억제되었다. 그러나 $Fe^{2+}$ /첨가 시에는 효소의 활성에 크게 영향을 미치지 않는 것으로 나타났다. NTR의 활성에 대한 억제물질의 영향은 $\beta$-mercaptoethanol 첨가 시에 효소의 활성이 모두 억제되었고, dithiothreitol, EDTA, 그리고 NaCl 첨가시에도 활성이 감소하는 것으로 확인되었다. TNT와 그 유사기질을 이용하여 HK-6에서 분리된 NTR의 기질 특이성을 조사한 결과, TNT, nitrobenzene, 그리고 RDX에 대해서는 비교적 활성이 높게 나타났으나 2,6-DNT와 2,4-DNT에서는 낮은 활성을 나타내는 것으로 확인되었다.