• Title/Summary/Keyword: acetyl-CoA synthetase

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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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    • v.29 no.4
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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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    • v.28 no.2
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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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Evience that a Plasmid Endoces Genes for Metabolism of Malonte in Pseudomonas fluorescens (Pseudomonas fluorescens에 있는 하나의 Plasmid가 말론산 대사에 관련된 유전자를 가지고 있다는 증거)

  • Kim, Yu-Sam;Kim, Eun-Joo
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.192-197
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    • 1994
  • Pseudomonas fluorescens which is able to utilize malonate as a sole carbon source was found to contain a novel 60 kb plasmid, which encodes the genes for the proteins to assimilate malonate, including malonate decarboxylase and acetyl-CoA synthetase. The evidence is as follows: The Pseudomonas cured with mitomycin C was unable to grow on malonate-medium as well as it lost plasmid. The plasmid isolated from the Pseudomonas could be introduced into E. coli strain JM103 and DH1 by transformation. The transformed E. coli was able to grow on malonate-medium and could transmit its plasmid back to the cured P. fluorescens by conjugation. The existence of the plasmid in the transformed E. coli was confirmed by hybridization with a labeled probe prepared from 12 kb segment of the plasmid. Dot hybridization showed that the copy number of the plasmid in the transformed E. coli is at least 13 times higher than in the wild type P. fluorescens. The two key enzymes, malonate decarboxylase and acetyl-CoA synthetase, were inducible by malonate in the transformed E. coli.

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

  • 김성준;김유삼
    • Korean Journal of Microbiology
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    • v.23 no.3
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    • pp.230-234
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    • 1985
  • A bacterium which can utilize malonate as a sole carbon source was isolated from soil. This strain was identified to be Acinetobacter calcoaceticus by morphological, cultural, phtsiological and biochemical examination. When this microorganism was grown on malonate as a aole carbon source, the enzymes, such as malonyl-CoA synthetase, isocitrate lyase and malate synthase were induced. These results suggest that in this microorganism, malonate is also assimilated through the proposed pathway in Pseudomonas fluorescens: $malonate{\rightarrow}malonyl-CoA{\rightarrow}acetyl-CoA{\rightarrow}glyoxylate\;cycle$.

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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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    • v.36 no.3
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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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    • v.5 no.1
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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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    • v.19 no.10
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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.

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

  • 김현수;정지천
    • The Journal of Korean Medicine
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    • v.23 no.4
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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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Classification According to Site of Action of Paddy Herbicides Registered in Korea (국내 수도용 제초제의 작용기작별 분류)

  • Park, Jae-Eup;Kim, Sang-Su;Kim, Young-Lim;Kim, Min-Ju;Ha, Heun-Young;Lee, In-Yong;Moon, Byung-Chul;Ihm, Yang-Bin
    • Weed & Turfgrass Science
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    • v.3 no.3
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    • pp.165-173
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    • 2014
  • This review study was conducted to recommend the effective use of herbicide mixtures in Korea. The herbicide ingredients by Herbicide Resistancce Action Committee (HRAC) was classified into 23 groupes according to the mode of action (acetyl CoA carboxylase inhibitors, acetolactate synthase, photosystem I and II inhibitors, protoporphyrinogen oxidase inhibitors, carotenoid biosynthesis inhibitors, enolpyruvyl shikimate-3-phosphate synthase inhibitors, glutamine synthetase inhibitors, dihydropteroate synthetase inhibitors, mitosis inhibitors, cellulose inhibitors, oxidative phosphorylation uncouplers, fatty acid and lipid biosynthesis inhibitors, synthetic auxins, auxin transport inhibitors and potential nucleic acid inhibitors or non-descript mode of action). The rice herbicide mixtures registered in Korea were classified based on the guideline of HRAC. Accordingly, such a classification system for resistance management can help to avoid continuous use of the herbicide having the same mode of action in the same field.