• 제목/요약/키워드: intracellular enzyme

검색결과 321건 처리시간 0.028초

Purification and Characterization of Soybean Cotyledonary Spermidine Dehydrogenase

  • Park, Sung-Joon;Cho, Young-Dong
    • BMB Reports
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    • 제28권5호
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    • pp.408-413
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    • 1995
  • Decrease in the amount of cotyledonary spermidine in Glycine max under anaerobic conditions related to an increase in spermidine dehydrogenase. Under the same conditions, no enzymatic activity of diamine oxidase was observed. Exposure of Glycine max both to spermidine and 1,3-diaminopropane under anaerobic conditions resulted in a decrease in spermidine contents. Correlated with the decrease in spermidine contents, there was a drastic increase in spermidine dehydrogenase. The molecular weight of the purified enzyme estimated by Sephacryl S-300 gel column and SDS gel electrophoresis were 130,000 dalton and 65,000 dalton, respectively, indicating that the enzyme is a dimer. The optimal pH for activity was 9.3. The $K_m$ value for spermidine was 0.61 mM. Neither metal ions nor polyamine and derivatives affected enzymatic activity, but the enzyme was inhibited by DTNB, NEM and PCMB, suggesting that a cysteine residue of the enzyme is associated with or involved in enzyme activity. To our knowledge, this is the first report describing properties of the enzyme from plants. Considered together, the data in this paper indicate that both spermidine and 1,3-diaminopropane, novel activators, enhance the spermidine dehydrogenase activity and control the intracellular spermidine contents.

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Saccharomyces cerevisiae에서 PNP의 부분 정제와 특성 (Partial Purification and Characterization of Purine Nucleoside Phosphorylase in Saccharomyces cerevisiae)

  • 최혜선
    • 미생물학회지
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    • 제29권3호
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    • pp.172-178
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    • 1991
  • Intracellular purine nucleoside phosphorylase (PNP) from Saccharomyces cerevisiae was partially purified using ammonium sulfate fractionation, heat treatment, a DEAE-Sephadex A-50 anion exchange chromatography and a Sephadex G-100 gel filtration chromatography. The enzyme was purified 20 fold with 3% recovery. The stability of enzyme was kept by addition of inosine and dithiothreitol. The pH optimum was found to be from 6.3 to 7.3 PNP was sensitive to 10mM of $Hg^{2+}$ , $Cu^{2+}$ , and was inactivated completely by 2 mM of p-chloromercuribenzoate and 5,5'-dithiobis (2-nitrobenzoate). The enzyme was capable of catalyzing the phosphorolysis of inosine, deoxyinosine, guanosine, deoxyguanosine and adenosine.

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Effect of glutathione on tetraploid embryo development in the pigs

  • Kim, Hwa-Young;Lee, Sang-Hee;Hwangbo, Yong;Lee, Seung Tae;Lee, Eunsong;Park, Choon-Keun
    • 한국수정란이식학회지
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    • 제31권3호
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    • pp.207-213
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    • 2016
  • The objective of this study was to investigate to influence of glutathione (GSH) on development and antioxidant enzyme activity in tetraploid porcine embryos. Tetraploid embryos were produced using parthenogenetic 2-cell embryo by electrofusion method. Tetraploid embryo development was observed every 24 hours and intracellular antioxidant enzyme activity was measured at 120 hours after electrofusion. The 4-cell to 16-cell stage tetraploid embryos was increased in 100 and $500{\mu}M$ GSH-treated groups compared control group at 48 hours (P < 0.05) but cleavage rates were not significantly different among the GSH treatment groups at 48, 72, 96, and 120 hours. Blastocyst formation was significantly increased by 300 and $500{\mu}M$ GSH at 120 hours in tetraploid embryos (P < 0.05). But blastocyst cell number were not significantly different among the GSH treatment groups ($16.4{\pm}0.8$, $16.8{\pm}2.6$, $18.5{\pm}2.8$ and $17.5{\pm}1.8$). The intracellular antioxidant enzyme level was increased in $500{\mu}M$ GSH compared to 0 and $100{\mu}M$ GSH (P < 0.05). We suggest that GSH may be improve development of tetraploid embryo in pigs.

고온성세균의 $\beta$-Galactosidase에 관한 연구( II )-효소의 생산, 정제 및 정제효소의 성질- (Studies on the $\beta$-Galactosidase from Thermophilic Bacterium - On the Production, Purification of Enzyme and the Properties of the Purified Enzyme -)

  • 오만진;이종수;김해중;김찬조
    • 한국미생물·생명공학회지
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    • 제11권1호
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    • pp.15-21
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    • 1983
  • 전보에서 분리 동정한 Y-33 균주의 $\beta$-galactosise 효소생산조건을 검토하고 효소를 정제하여 정제효소의 성질을 조사한 결과는 다음과 같다. 1. 효소생산을 위한 최적초발 pH는 7.0이었고 최적온도는 $65^{\circ}C$이었다. 2. 효소는 lactose와 galactose에 의하여 유도되어졌으며 세포내효소이었다. 3. 조효소액을 1차 DEAE-cellulose, 2차 DEAE-cellulose column chromatography 및 Sephadex G-150로 gel filtration하여 정제도가 28.3배, 수율이 15.2%의 정제효소를 얻었다. 4. 정제효소는 polyacrylamide gel 전기 영동에 의하여 순도가 검정되었다. 5. 정제효소의 유당가수분해를 위한 최적작용 온도는 $65^{\circ}C$. pH는 6.5이었다.

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Hydrogen peroxide attenuates refilling of intracellular calcium store in mouse pancreatic acinar cells

  • Yoon, Mi Na;Kim, Dong Kwan;Kim, Se Hoon;Park, Hyung Seo
    • The Korean Journal of Physiology and Pharmacology
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    • 제21권2호
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    • pp.233-239
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    • 2017
  • Intracellular calcium ($Ca^{2+}$) oscillation is an initial event in digestive enzyme secretion of pancreatic acinar cells. Reactive oxygen species are known to be associated with a variety of oxidative stress-induced cellular disorders including pancreatitis. In this study, we investigated the effect of hydrogen peroxide ($H_2O_2$) on intracellular $Ca^{2+}$ accumulation in mouse pancreatic acinar cells. Perfusion of $H_2O_2$ at $300{\mu}M$ resulted in additional elevation of intracellular $Ca^{2+}$ levels and termination of oscillatory $Ca^{2+}$ signals induced by carbamylcholine (CCh) in the presence of normal extracellular $Ca^{2+}$. Antioxidants, catalase or DTT, completely prevented $H_2O_2$-induced additional $Ca^{2+}$ increase and termination of $Ca^{2+}$ oscillation. In $Ca^{2+}$-free medium, $H_2O_2$ still enhanced CCh-induced intracellular $Ca^{2+}$ levels and thapsigargin (TG) mimicked $H_2O_2$-induced cytosolic $Ca^{2+}$ increase. Furthermore, $H_2O_2$-induced elevation of intracellular $Ca^{2+}$ levels was abolished under sarco/endoplasmic reticulum $Ca^{2+}$ ATPase-inactivated condition by TG pretreatment with CCh. $H_2O_2$ at $300{\mu}M$ failed to affect store-operated $Ca^{2+}$ entry or $Ca^{2+}$ extrusion through plasma membrane. Additionally, ruthenium red, a mitochondrial $Ca^{2+}$ uniporter blocker, failed to attenuate $H_2O_2$-induced intracellular $Ca^{2+}$ elevation. These results provide evidence that excessive generation of $H_2O_2$ in pathological conditions could accumulate intracellular $Ca^{2+}$ by attenuating refilling of internal $Ca^{2+}$ stores rather than by inhibiting $Ca^{2+}$ extrusion to extracellular fluid or enhancing $Ca^{2+}$ mobilization from extracellular medium in mouse pancreatic acinar cells.

Polyamine과 Polyamine의 생합성에 관련된 효소들의 시금치잎 세포내 분포에 관한 연구 (Studies on the Intracellular Localization of Polyamines and Their related Enzymes in Spinach Leaves)

  • 김성호
    • Journal of Plant Biology
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    • 제32권4호
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    • pp.285-292
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    • 1989
  • The intracellular localizations of polyamines and their related enzymes were investigated from young spinach leaves. Polyamines were present in all parts of plant cells, both in the subcellular organelles and in the soluble fraction of cytoplasm, however, polyamines were mainly located in the cytosolic fraction. Most activities of L-arginine decarboxylase(ADC) and L-ornithine decarboxylase(ODC), two important enzymes of putrescine and polyamine biosynthesis, were detected in cytosol fraction, while in subcellular organelles the activities were very low. Activities of diamine oxidase(DAO) and polyamine oxidase(PAO), the catabolic enzyme of diamine and polyamine, were not detected in spinach leaves. It was suggested that polyamines and their related synthetic enzymes were located in the soluble fraction of cytoplasm.

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Induction of Quinone Reductase , an Anticarcinogenic Marker Enzyme, by Vitamin E in Both Hepalclc7 Cells and Mice

  • Kwon, Chong-Suk;Kim, Jong-Sang
    • Preventive Nutrition and Food Science
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    • 제4권2호
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    • pp.122-124
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    • 1999
  • Induction of NAD(P)H : (quinone-acceptor) oxidoreductase (QR) which obligatory two electron reduction of quinones and prevents their participation in oxidative cycling and thereby the depletion of intracellular glutathione, has been used as a marker for chemopreventive agents. We postulated that vitamin E, an antioxidant, which induces QR as the gene of QR was reported to contain antioxidant reponsive element in the 5'-flanking region. Vitamin E resulted in significant induction of QR in both hepalclc7 cells and mouse tissues. QR induction was observed; to be maximal at 25uM vitamin E for hepalclc7 cells while it was maximal in the level of 2.5∼5 μmoles vitamin E/㎏ BW for mouse tissues. Thus the cancer-preventive effect of vitamin E may be exerted by it induction of intracellular detoxifying enzymes.

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Bacillus subtilis가 생산하는 비특이적 $\beta$-fructofuransoidase의 부분정제 및 특성 (Partial Purification and Properties of Non-specific $\beta$ -fructofuranosidase Produced by Bacillus subtilis)

  • 송근섭;엄태붕
    • 한국미생물·생명공학회지
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    • 제18권5호
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    • pp.484-489
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    • 1990
  • Bacillus subtilis의 세포내 이눌라아제가 부분정제되고 그의 작용 모드와 일반적 특성이 조사되었다. 이 효소는 gel filtration에 의하여 분자량을 추정하였을 때 49,000이었고, 등진점은 5.2 이었다. 기질에 대한 친화성의 지표인 Km값은 설탕에 대해서는 10mM, 라피노오스에 대해서는 18mM 이었다. 이 효소는 산성쪽에서는 불안정한 단백질로서 pH6.6에서 최대 활성을 보였으며 최적온도는 10분간 반응시켰을 때 50'C였다. 이 효소의 작용모든는 이눌린같은 구조를 가지는 과당 중합체를 과당 끝부분으로부터 하나씩 잘라가는 exo-cleavage 형이었다.

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Aureobasidium pullulans C-23 균주에 의한 Fructosyl Transferase의 생산 최적 배양조건 (Optimum Culture Conditions for the Production of Fructosyl transferase by Aureobasidium pullulans C-23)

  • 조원태;임재윤
    • 한국미생물·생명공학회지
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    • 제18권4호
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    • pp.417-422
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    • 1990
  • 선발균의 효소생산성 향상을 위해 배양과정의 특징을 조사하였다. 효소생산을 위한 최적의 탄소원은 sucrose (35)이고, 질소원은 ammonium oxalate와 yeast extract를 함께 넣어 주었을 때이었다. 5l jar fermentor에서 선발균의 배양형태를 조사한 결과 2-3일 배양기간에는 pH가 4 이하로 현저하게 감소하는 것을 관찰하였고 4일 이후 다시 pH가 5-6 사이로 증가하였다. 균의 성장은 4일까지 증가하였다. 균체내 효소의 생성은 2일까지는 현저한 증가를 보이고 3-4일 정도 큰 변화를 나타내지 않으며 5일 이후 감소하였다. 균체외 효소는 5-6 일까지 계속 증가하는 형태를 나타내었다. Glucose로 24시간 동안 미리 배양한 후 surcose를 첨가하여 배양했을 때 균체내 효소의 생산이 현저히 증가하는 것을 알 수 있었다.

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Purification and Characterization of Manganese Superoxide Dismutase from Staphylococcus sciuri

  • Song, Chi-Hyun;Park, Eun-Kyung;Suh, Hyung-Joo;Lee, Yong-Se;Choi, Jang-Won;Ra, Kyung-Soo
    • Journal of Microbiology and Biotechnology
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    • 제9권3호
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    • pp.271-275
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    • 1999
  • The intracellular superoxide dismutase (SOD) from Staphylococcus sciuri was isolated to homogeneity by continuous steps, including ammonium sulfate fractionation, DEAE-ion-exchange chromatography, gel filtration, and phenyl hydrophobic gel chromatography. Pure SOD had a specific activity of 4,625 U/mg and was purified 158-fold with a yield of 31 % from a cell free extract. The molecular weight of the purified SOD was determined to be approximately 35.5 kDa by gel filtration and the enzyme was also shown to be composed of dimeric subunits on denaturing SDS-PAGE. The enzyme activity remained stable at pH 5 to 11 and also to heat treatment of up to $50^{\circ}C$ at pH 7.8, with 80% relative activity. The enzyme was insensitive to cyanide, hydrogen peroxide, and azide, indicating that it is a manganese-containing SOD. The EPR spectrum showed the enzyme containing manganese as a cofactor.

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