• 제목/요약/키워드: Aspartate aminotransferase activity

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간질(Fasciola hepatica)의 Aspartate 및 Alanine Aminotransferase에 관하여 (Aspartate and Alanine Aminotransferase in Fasciola hepatica)

  • 박선효;권년수이희성송철용
    • 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.

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Streptomyces fradiae에서 분리된 Aspartate Aminotransferase의 특성 (Characterization of Aspartate Aminotransferase Purified from Streptomyces fradiae)

  • Lee, Sang-Hee;Lee, Kye-Joon
    • 미생물학회지
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    • 제31권3호
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    • pp.237-244
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    • 1993
  • Aspartate aminotransferase (ASAT) (L-aspartate : 2-oxyoglutarate, EC 2.6. 1. 1.) from Streptomyces fradiae NRRL 2702 has been purified by acetone precipitation, DEAE-cellulose, hydroxyapatite, and preparative electrophoresis (Prep cell), of which the last was the most effective step in the purification of ASAT. The molecular mass was estimated to be 54,000 dalton by SDS-PAGE and 120,000 dalton by gel filtration chromatography. Preparative isoelectric focusing of purified ASAT resulted in one polypeptide band with a pI of 4.2, showing homogeneity and indicating that the enzyme is composed of two identical subunits. The enzyme was specific for L-aspartate as an amino donor ; the $K_{m}$ values were determined to be 2.7 mM for L-aspartate, 0.7 mM for 2-oxoglutarate, 12.8 mM for L-glutamate, and 0.15 mM for oxaloacetate. The enzyme was relatively heat-stable, having maximum activity at 55.deg.C, and it had a broad pH optimum ranging from 5.5 to 8.0. The activity of the purified enzyme was not inhibited by ammonium ions. This paper reports the first purification and characterization of the aspartate aminotransferase from a species of Streptomyces.s.

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Xanthomonas oryzae pv. oryzae로 부터 aspartate aminotransferase 유전자의 분리 및 생화학 특성 (Cloning and Biochemical Characterization of Aspartate Aminotransferase from Xanthomonas oryzae pv. oryzae)

  • 강한철;윤상홍;이창묵
    • Journal of Applied Biological Chemistry
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    • 제52권3호
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    • pp.109-115
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    • 2009
  • Xoo로 부터 aspartate aminotransferase로 추정되는 유전자를 분리한 다음 발현시켜 생화학 특성을 조사하였다. 분리된 유전자는 His6 pET-21(a) 운반체에 삽입시켰으며 E. coli BL21(DE3)에서 발현시켰다. 재조합된 Asp-AT는 affinity chromatography를 이용하여 분리하였으며 SDS-PAGE분석에서 43kDa의 단일 밴드를 나타내었다. 분리된 효소는 amino donor 로서 L-aspartate에 대하여 효소활성도가 가장 높았고, L-leucine 및 L-cysteine에 대하여서도 상당한 활성도를 나타내었다. 효소의 최적 pH는 약 7.5 근처에서 나타났고 효소의 안정성은 산성조건 보다는 알칼리 조건에서 훨씬 높았다. 최적 온도는 약 $35-40^{\circ}C$로 나타났고 $55^{\circ}C$에서 20분간 열처리한 이후의 잔여 활성도는 약 78%로 나타났다. 여러 중금속 중에서 망간 이온에 의해 효소활성이 촉진되었다.

Effects of Pyridoxine on Growth Performance and Plasma Aminotransferases and Homocysteine of White Pekin Ducks

  • Xie, Ming;Tang, Jing;Wen, Zhiguo;Huang, Wei;Hou, Shuisheng
    • Asian-Australasian Journal of Animal Sciences
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    • 제27권12호
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    • pp.1744-1748
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    • 2014
  • A dose-response experiment with seven supplemental pyridoxine levels (0, 0.66, 1.32, 1.98, 2.64, 3.30, and 3.96 mg/kg) was conducted to investigate the effects of pyridoxine on growth performance and plasma aminotransferases and homocysteine of White Pekin ducks and to estimate pyridoxine requirement for these birds. A total of 336 one-day-old male White Pekin ducks were divided to 7 experimental treatments and each treatment contained 8 replicate pens with 6 birds per pen. Ducks were reared in raised wire-floor pens from hatch to 28 d of age. At 28 d of age, the weight gain, feed intake, feed/gain, and the aspartate aminotransferase, alanine aminotransferase, and homocysteine in plasma of ducks from each pen were all measured. In our study, the pyridoxine deficiency of ducks was characterized by growth depression, decreasing plasma aspartate aminotransferase activity and increasing plasma homocysteine. The ducks fed vitamin $B_6$-deficient basal diets had the worst weight gain and feed/gain among all birds and this growth depression was alleviated (p<0.05) when pyridoxine was supplemented to basal diets. On the other hand, plasma aspartate aminotransferase and homocysteine may be the sensitive indicators for vitamin $B_6$ status of ducks. The ducks fed basal diets had much lower aspartate aminotransferase activity and higher homocysteine level in plasma compared with other birds fed pyridoxine-supplemented diets (p<0.05). According to quadratic regression, the supplemental pyridoxine requirements of Pekin ducks from hatch to 28 days of age was 2.44 mg/kg for feed/gain and 2.08 mg/kg for plasma aspartate aminotransferase and the corresponding total requirements of this vitamin for these two criteria were 4.37 and 4.01 mg/kg when the pyridoxine concentration of basal diets was included, respectively. All data suggested that pyridoxine deficiency could cause growth retardation in ducks and the deficiency of this vitamin could be indicated by decreasing plasma aspartate aminotransferase activity and increasing plasma homocysteine.

장기간의 Pyridoxine 부족이 쥐 간의 Mitochondria 및 Cytosolic Fraction 에 있는 Aspartate Aminotransferase 및 Pyridoxal 5'- Phoshate에 미치는 영향 (Effect of Long-Term Pyridoxine Depletion on Asparte Aminotransferase and Pyridoxal 5'-Phosphate of Rat Liver Mitochondrial and Cytosolic Fractions)

  • 임경숙
    • Journal of Nutrition and Health
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    • 제19권4호
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    • pp.246-254
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    • 1986
  • 이유한 Sprague Dawley 종의 암컷 흰쥐를 두군으로 나누어 각각 pyridoxine이 충분한 식이 (22mg/kg diet)와 pyridoxine이 부족한 식이 (1.2mg/kg diet0로 성장시킨후, 적절한 시기에 임신시켰다. 태어난 새끼가 이유할 때 결핍식이군을 둘로 나누어, 한쪽은 pyridoxine이 충분한 식이로 바꾸어준후, 세군의 쥐를 생후 10-주까지 성장시켰다. 각 군의 새끼쥐가 각각 0, 3, 7, 10 주 되었을대, 간의 mitochondria 및 cytosolic fraction 에 있는 Asparate aminotransferase [ EC 2.6.1.1] activity 및 pyridoxal phosphate의 함량을 측정하였다. 결핍군의 간 aspartate aminotransterase activity는 mitochondria 및 cytosolic fraction에서 각각 대조군에 비해 유의적으로 낮았으며 10-4 M pyridoxal phosphate를 첨가한 후 측정한 total enzyme activity 는 거의 대조군과 비슷하였다. 3 주이후 pyridoxine 이 충분한 식이로 바구어준 회복군에서도 대조군에 비해 유의적으로 낮았으나, 결핍군보다는 높았다. 이두 fraction 에서의 pyridoxal phosphate 함량은 결핍군이 대조군보다 현저히 낮았으며, 회복군은 대조군과 비슷하였다. 이로써, 장기간에 걸친 pyridoxine결핍이간의 aspartate aminotransferase activity 및 pyridoxal phosphate에 현저한 영향을 미치며 이유기 이후의 식이보충에 의해서도 enzyme activity는 쉽게 회복되지 않음을 알 수있다.

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Shifts in Protein Metabolism in Hemolymph and Fat Body of the Silkworm, Bombyx mori L. in Response to Fluoride Toxicity

  • Ramakrishna, S.;Jayaprakash, Jayaprakash
    • International Journal of Industrial Entomology and Biomaterials
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    • 제15권1호
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    • pp.59-68
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    • 2007
  • Changes in protein metabolism were studied in hemolymph and fat body on days 1, 3, 5 and 7 of the fifth-instar silkworm, Bombyx mori, exposed to lethal, sublethal doses and prevailing levels of fluoride in groundwater in Karnataka and Andhra Pradesh States of India. The total protein content indicated a depletion followed by a concomitant increase in accumulation of free amino acids. Concurrently, the activity of protease in both of the tissues was also increased. A steady enhancement in the activities of alanine aminotransferase and aspartate aminotransferase paralleled the elevation of glutamate dehydrogenase activity in the tissues studied. It is presumed, on the basis of these results, that the fluoride toxicity causes major changes in protein metabolism of the silkworms.

사염화탄소에 의한 간손상에 미치는 고본의 보호작용 (Protective Effects of Angelica tenuissima Nakai on Hepatotoxicity by Carbon Tetrachloride in Rats)

  • 정춘식;정기화
    • Biomolecules & Therapeutics
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    • 제10권4호
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    • pp.211-217
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    • 2002
  • Hepatoprotective activity of methanol extract of Angelica tenuissima Nakai on the $CCl_4$-induced hepatotoxicity was investigated. To elucidate the hepatoprotective activity and free radical scavenging effect, we examined alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin, total protein, cholesterol, malondialdehyde (MDA) levels in serum and activities of superoxide dismutase (SOD), catalase (CAT) in hepatic tissue as compared with those of carbon tetrachloride-induced rats. The action mechanism also has been estimated by quantative analysis of cytochrome P450 (CYP), NADPH-CYP reductase for phase I metabolism and glutathion (GSH), glutathion S-transferase (GST) level for phase II metabolsim. Treatment of Angelica tenuissima methanol extract significantly lowered the levels of alanine aminotransferase and aspartate aminotransferase. In addition, the levels of cholesterol, triglyceride, MDA, CAT were decreased, and SOD was activated. This result indicates that the hepatoprotective effect of Angelica tenuissima methanol extract on the CCl4-induced hepatotoxicity would be originated from reduction of the NADPH-CYP reductase, GSH and the enhancement of the activities of GST, CYP.

사염화탄소로 유발된 간손상에서의 효소 활성도의 변화로 본 홍화자 분획물의 간손상 보호 작용 (Hepatoprotective Effect of Subfractions of Carthamus tinctorius L. Semen on the Reversal of Biotransformation Enzyme Activities in CCl4-induced Hepatotoxic Rats)

  • 정춘식;정기화;정정숙
    • 한국식품위생안전성학회지
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    • 제14권2호
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    • pp.172-178
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    • 1999
  • Previous studies have shown that methanol extract and its butanol fraction of Carthamus tinctorius L. Semen have the hepatoprotective effect on the CCl4-induced hepatotoxicity. The hepatoprotective effect of subfractions has been evaluated by analyzing blood and hepatocyte biochemical analyses and biotransformation enzyme analyses. Treatment of BS-5, subfraction has significantly decreased the activities of alanine aminotransferase and aspartate aminotransferase. In addition, the levels of cholesterol and triglyceride in liver have been decreased as compared with that of CCl4 treated rats. The hepatoprotective effect of BS-5, subfraction on the CCl4-induced hepatotoxicity would be mediated of the attenuation of the level of cytochrome P450 and the enhancement of the activity of glutathion S-transferase.

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Aspartate aminotransferase activity in the pulp of teeth treated for 6 months with fixed orthodontic appliances

  • Veberiene, Rita;Latkauskiene, Dalia;Racinskaite, Vilma;Skucaite, Neringa;Machiulskiene, Vita
    • 대한치과교정학회지
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    • 제45권5호
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    • pp.261-267
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    • 2015
  • Objective: To measure aspartate aminotransferase (AST) activity in the pulp of teeth treated with fixed appliances for 6 months, and compare it with AST activity measured in untreated teeth. Methods: The study sample consisted of 16 healthy subjects (mean age $25.7{\pm}4.3$ years) who required the extraction of maxillary premolars for orthodontic reasons. Of these, 6 individuals had a total of 11 sound teeth extracted without any orthodontic treatment (the control group), and 10 individuals had a total of 20 sound teeth extracted after 6 months of orthodontic alignment (the experimental group). Dental pulp samples were extracted from all control and experimental teeth, and the AST activity exhibited by these samples was determined spectrophotometrically at $20^{\circ}C$. Results: Mean AST values were $25.29{\times}10^{-5}U/mg$ (standard deviation [SD] 9.95) in the control group and $27.54{\times}10^{-5}U/mg$ (SD 31.81) in the experimental group. The difference between these means was not statistically significantly (p = 0.778), and the distribution of the AST values was also similar in both groups. Conclusions: No statistically significant increase in AST activity in the pulp of mechanically loaded teeth was detected after 6 months of orthodontic alignment, as compared to that of teeth extracted from individuals who had not undergone orthodontic treatment. This suggests that time-related regenerative processes occur in the dental pulp.