• 제목/요약/키워드: acetyl-CoA synthetase

검색결과 31건 처리시간 0.028초

Properties of Acetyl-CoA Synthetase from Pseudomonas fluorescens

  • Kim, Yu-Sam;An, Jae-Hyung;Yang, Bu-Hyun;Kim, Kyu-Wan
    • BMB Reports
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    • 제29권4호
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    • pp.277-285
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    • 1996
  • In Pseudomonas fluorescens grown on malonate as sole carbon source, acetyl-CoA synthetase was induced, suggesting that malonate is metabolized through acetate and then acetyl-CoA. Acetyl-CoA synthetase was purified 18.6-fold in 4 steps to apparent homogeneity. The native molecular mass of the enzyme estimated by a native acrylamide gel electrophoresis was 130 kDa. The enzyme was composed of two identical subunits with a molecular mass of 67 kDa. Optimum pH was 70. The acetyl-CoA synthetase showed typical Michaelis-Menten kinetics for the substrates, acetate, ATP and CoA, whose $K_m$ values were calculated to be 33.4, 74.8, and 40.7 mM respectively. Propionate. butyrate and pentanoate were also used as substrates by the enzyme, but the rate of the formation of the CoA derivatives was decreased in the order of the increase in carbon number. The enzyme was inhibited by the group-specific reagents diethylpyro-carbonate, 2,3-butanedione, pyridoxal-5'-phosphate and N-bromosuccinimide. In the presence of substrates the inactivation rate of the enzyme, by all of the group-specific reagents mentioned above decreased, indicating the presence of catalytically essential histidine, arginine, lysine and tryptophan residues at or near the active site. Preincubation of the enzyme with ATP, $Mg^{2+}$ resulted in the increase of its susceptibility to diethylpyrocarbonate, suggesting that ATP, $Mg^{2+}$ may induce a conformational change in the active site exposing the essential histidine residue to diethylpyrocarbonate. The enzyme was acetylated in the presence of acetyl-CoA, indicating that this is one of acyl-enzyme.

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Metabolic Routes of Malonate in Pseudomonas fluorescens and Acinetobacter calcoaceticus

  • Byun, Hye-Sin;Kim, Yu-Sam
    • BMB Reports
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    • 제28권2호
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    • pp.107-111
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    • 1995
  • In malonate grown Pseudomonas fluorescens, malonate decarboxylase and acetyl-CoA synthetase were induced, whereas in Acinetobacter calcoaceticus malonate decarboxylase, acetate kinase, and phosphate acetyltransferase were induced. In both bacteria malonate decarboxylase was the first, key enzyme catalyzing the decarboxylation of malonate to acetate, and it was localized in the periplasmic space. Acetate thus formed was metabolized to acetyl-CoA directly by acetyl-CoA synthetase in Pseudomonas, and to acetyl-CoA via acetyl phosphate by acetate kinase and phosphate acetyltransferase in Acinetobacter.

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Pseudomonas fluorescens에 있는 하나의 Plasmid가 말론산 대사에 관련된 유전자를 가지고 있다는 증거 (Evience that a Plasmid Endoces Genes for Metabolism of Malonte in Pseudomonas fluorescens)

  • 김유삼;김은주
    • 미생물학회지
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    • 제32권3호
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    • pp.192-197
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    • 1994
  • 말론산을 탄소원으로 하여 성장하는 Pseudomonas fluorescens에서 60 kb 정도의 plasmid를 발견하였다. 이 plasmid를 curing한 Pseudomonas는 malonate 배지에서 자라지 못하였고 plasmid도 소실되었다. 또한 이 plasmid를 transformation과 conjugation으로 각각 E. coli와 cured P. fluorescens로 이동시킨 결과 이 plasmid를 받은 transformed E. coli와 conjugant P. fluorescens는 malonate를 탄소원으로 하여 성장하였고 malonate 대사에 관련된 효소인 malonate decarboxylase와 acetyl-CoA synthetase의 활성이 측정되었다. Western blotting을 통하여 tansformed E. coli에서 P. fluorescens와 동일한 acetyl-CoA synthetase가 malonate에 의해 induction됨을 확인하였다.

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토양으로부터 Malonate를 이용하는 Acinetobacter calcoaceticus의 분리 (Isolation of a Malonate-utilixing Acinetobacter calcoaceticus from Soil)

  • 김성준;김유삼
    • 미생물학회지
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    • 제23권3호
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    • pp.230-234
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    • 1985
  • Malonate를 유일한 탄소원으로 활용할 수 있는 세균을 토양으로부터 분리하였다. 이 세균은 행태, 배양, 생리 그리고 생화학적 연구를 통하여 Acinetobacter calcoaceticus임이 확인되었다. 이 미생물을 malonate를 유일한 탄소원으로 하는 배지에서 배양하였을 갱우, malonyl CoA synthetase, isocitrate lyase 및 malate, synthase가 유도 되었다. 따라서 이 미생물에서도 Pseudomonas fluorescens에서 제안되었던 대사경로 즉 $malonate{\rightarrow}malonyl-CoA{\rightarrow}acetyl-CoA{\rightarrow}glyoxylate\;cycle$을 통하여 malonate를 이용하는 것으로 판단된다.

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The Effects of a High-fat or High-sucrose Diet on Serum Lipid Profiles, Hepatic Acyl-CoA Synthetase, Carnitine Palmitoyltransferase-I, and the Acetyl-CoA Carboxylase mRNA Levels in Rats

  • Ryu, Mi-Hyun;Cha, Youn-Soo
    • BMB Reports
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    • 제36권3호
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    • pp.312-318
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    • 2003
  • The purpose of this study was to investigate the effects of altering relative intakes of fat and carbohydrates on serum lipid profiles, hepatic acyl-CoA synthetase (ACS), carnitine palmitoyltransferase-I (CPT-I), and the acetyl-CoA carboxlyase (ACC) mRNA level in Sprague-Dawley rats. For four weeks the rats were fed either an AIN-76 diet or one of its modified diets that were supplemented with 20% beef tallow (high-fat diet, HF) and 66.3% sucrose (highsucrose diet, HS). The HS group had significantly higher serum triglyceride and total cholesterol concentrations when compared with the other groups. Serum LDL-cholesterol concentrations in the HS and HF groups were significantly higher when compared to the normal diet (ND) group. Serum HDL-cholesterol levels of the ND and HS groups were significantly higher than those of the HF group. The hepatic total lipid level of the HF group was significantly higher than those of other groups; triglyceride levels of the HS and HF groups were significantly higher than those of the ND group. Hepatic ACS mRNA levels of the HF group were significantly higher than those of the ND group. Hepatic CPT-I mRNA levels were higher in the HF group than other groups. Also, ACC mRNA levels in the liver increased in the HF group. In conclusion, changes in the composition of dietary fat and carbohydrates could affect the hepatic ACS, CPT-I, and ACC mRNA levels. These results facilitate our understanding of the coordinated regulation of the ACS, CPT-I, and ACC mRNA levels and will serve to enhance our understanding of the molecular mechanisms that underlie the regulation of fatty acid metabolism.

EFFECTS OF DIETARY PROTEINS ON THE ACTIVITIES OF LIPOGENIC ENZYMES IN THE LIVER OF GROWING CHICKS

  • Tanaka, K.;Okamoto, T.;Ohtani, S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제5권1호
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    • pp.123-128
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    • 1992
  • In Experiment 1, when fasted chicks were fed diets containing various sources of protein for 3 days, the activities of lipogenic enzymes (acetyl-CoA carboxylase, fatty acid synthetase, citrate cleavage enzyme and malic enzyme) in the liver of growing chicks were significantly lower in the soybean protein or gluten diet than in the casein or fish protein diet. Triglycride contents of the liver and plasma of chicks fed the casein or fish protein diet were significantly lower than that of those fed soybean protein or gluten diet. In Experiment 2, the effects of dietary amino acid mixture simulating casein or protein on the activities of hepatic lipogenic enzymes were examined. The activities of acetyl-CoA carboxylase and fatty acid synthetase in the liver of chicks fed the casein diet were significantly higher than that of those fed the soybean protein diet or two diets of amino acid mixtures. Furthermore, there were no significant differences between the two diets of amino acid mixture based on casein or soybean protein. However, the activities of malic enzyme and citrate cleavage enzyme tended to be lower in the soybean-type amino acid diet than in the casein-type amino acid diet. Thus, some effects can be ascribed to the protein itself and some to the amino acid composition of the protein sources.

Acetate Consumption Activity Directly Determines the Level of Acetate Accumulation During Escherichia coli W3110 Growth

  • Shin, Soo-An;Chang, Dong-Eun;Pan, Jae-Gu
    • Journal of Microbiology and Biotechnology
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    • 제19권10호
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    • pp.1127-1134
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    • 2009
  • Escherichia coli excretes acetate during aerobic growth on glycolytic carbon sources, which has been explained as an overflow metabolism when the carbon flux into the cell exceeds the capacity of central metabolic pathways. Nonacetogenic growth of E. coli on gluconeogenic carbon sources like succinate or in carbon-limited slow growth conditions is believed an evidence for the explanation. However, we found that a strain defected in the acs (acetyl Co-A synthetase) gene, the product of which is involved in scavenging acetate, accumulated acetate even in succinate medium and in carbon-limited low growth rate condition, where as its isogenic parental strain did not. The acs promoter was inducible in noncatabolite repression condition, whereas the expression of the ackA-pta operon encoding acetate kinase and phosphotransacetylase for acetate synthesis was constitutive. Results in this study suggest that E. coli excretes and scavenges acetate simultaneously in the carbon-limited low growth condition and in nonacetogenic carbon source, and the activity of the acetate consumption pathway directly affects the accumulation level of acetate in the culture broth.

상지추출물이 고지방식이에 의한 체중 변화와 지질대사에 미치는 영향 (Effects of Ramulus mori Extract on Obesity and Lipid Metabolism in High Fat Diet Rats)

  • 김현수;정지천
    • 대한한의학회지
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    • 제23권4호
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    • pp.64-72
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    • 2002
  • Objective: Ramulus mori (RM) has been known to be effective for the treatment of obesity. To show the effectiveness of RM in a more scientific way, RM extract was prepared and evaluated in high fat diet rats by measuring the changes of body weight and lipid metabolism as described briefly below. Methods: 200 g of crushed RM was extracted withmethyl alcohol. The extract was evaporated under reduced pressure to give 33.4 g. For 10 weeks, control group rats were fed a high fat diet, while the test group rats were fed with the same diet plus RM extract. The normal group was fed with a normal diet. 150 mg of RM extract per 1 kg of body weight was added to the diet in the test group rats. Results: The control group rats on the high fat diet gained weight significantly, whereas the test group rats on the high fat diet plus RM extract gamed less weight. The significant increase of liver weight caused by the high fat diet was also inhibited by the RM extract treatment. Total lipid, triglyceride and total cholesterol levels of serum in the high fat diet rats were remarkably increased, whereastheir levels on the high fat diet plus RM extract were less increased. While serum HDL-cholesterol levels were remarkably decreased in the high fat diet, its level was less decreased in the high fat diet plus RM extract. Furthermore, we observed that the activities of hepatic acetyl-CoA carboxylase and fatty acid synthetase increased under the high fat diet, while their activities under the high fat diet plus RM extract were getting back nearly to the normal levels of the normal diet rats. Conclusions: These result show that the obesity caused by a high fat diet was effectively inhibited by an RM extract. Our results also showed that the abnormal lipid metabolism caused by a high fat diet was effectively cured by adding RM extract.

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국내 수도용 제초제의 작용기작별 분류 (Classification According to Site of Action of Paddy Herbicides Registered in Korea)

  • 박재읍;김상수;김영림;김민주;하헌영;이인용;문병철;임양빈
    • Weed & Turfgrass Science
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    • 제3권3호
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    • pp.165-173
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    • 2014
  • 국내에 등록된 제초제의 효율적 사용을 위해서 제초제저항성관리위원회에서 제시한 제초제 작용기작별 분류를 기초로 23그룹으로 분류하였다. 세부그룹으로는 acetyl CoA carboxylase 억제제, acetolactate synthase 억제제, photosystem과 억제제, protoporphyrinogen oxidase 억제제, carotenoid biosynthesis 억제제, enolpyruvyl shikimate-3-phosphate synthase 억제제, glutamine synthetase 억제제, dihydropteroate synthetase 억제제, 세포분열 저해제(mitosis inhibitors), cellulose 생합성억제제, oxidative phosphorylation uncouplers, 지방산 및 지질생합성 억제제, synthetic auxins, auxin transport 억제제, potential nucleic acid Inhibitors 혹은 non-descript mode of action으로 나누었다. 이와 같은 그룹화 기준을 토대로 국내 등록되어 사용중인 논 제초제 성분들을 그룹화 하였다. 따라서, 이러한 약제 작용기작 관련 정보를 농약사용자에게 제공함으로써 특정약제의 연용과 중복사용을 방지하여 국내에서 제초제 저항성잡초 발생 문제를 줄일 수 있을 것으로 기대된다.