• Title/Summary/Keyword: Pyruvate carboxylase

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Effects of Cordyceps militaris on Key Enzymes of Carbohydrate Metabolism (Cordyceps militaris 성분이 당대사 관련 효소 활성에 미치는 영향)

  • Kim, Hyun-Sook;Ro, Young-Joo;Choe, Myeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.10
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    • pp.1531-1535
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    • 2005
  • The present study was designed to investigate the antidiabetic effect of Cordyceps militaris on activities of hepatic glucose-regulating enzymes, such as glucokinase (GK), pyruvate dehydrogenase (PDH) and acetyI-CoA carboxylase (ACC). We have isolated the active compounds, CM-A and CM-B from C. militaris and the extracts are under investigation to determine their mechanism of action. Hepatic GK, PDH and ACC activities were significantly (p < 0.05) increased compared with the control. Treatment with CM-A led to a rise in percentage of enzymes by 380$\%$,396$\%$ and 286$\%$, respectively, relative to control levels. CM-B were 329$\%$, 312$\%$ and 239$\%$, respectively. The increase in GK and ACC activities was linearly proportional with increased ratio of CM-A. Our findings suggest that C. militaris exerted antihyperglycemic potency, which is thought to be mediated through activation of GK, PDH and ACC enzymes related to glucose metabolism directly or indirectly and therefore C. militaris is promising as antidiabetic functional foods.

Discrimination of Bacillus subtilis from Other Bacillus Species Using Specific Oligonucleotide Primers for the Pyruvate Carboxylase and Shikimate Dehydrogenase Genes

  • Lee, Gawon;Heo, Sojeong;Kim, Tao;Na, Hong-Eun;Park, Junghyun;Lee, Eungyo;Lee, Jong-Hoon;Jeong, Do-Won
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.1011-1016
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    • 2022
  • Bacillus subtilis is a useful bacterium in the food industry with applications as a starter strain for fermented food and as a probiotic. However, it is difficult to discriminate B. subtilis from other Bacillus species because of high phenotypic and genetic similarity. In this study, we employed five previously constructed multilocus sequence typing (MLST) methods for the discrimination of B. subtilis from other Bacillus species and all five MLST assays clearly distinguished B. subtilis. Additionally, the 17 housekeeping genes used in the five MLST assays also clearly distinguished B. subtilis. The pyruvate carboxylase (pyrA) and shikimate dehydrogenase (aroE) genes were selected for the discrimination of B. subtilis because of their high number of polymorphic sites and the fact that they displayed the lowest homology among the 17 housekeeping genes. Specific primer sets for the pyrA and aroE genes were designed and PCR products were specifically amplified from B. subtilis, demonstrating the high specificity of the two housekeeping genes for B. subtilis. This species-specific PCR method provides a quick, simple, powerful, and reliable alternative to conventional methods in the detection and identification of B. subtilis.

Biotin Requirement for the Growth and Sporulation of Bacillus subtilis SNU816 in a Synthetic medium (Bacillus subtilis SNU816의 合成培地에서의 성장과 포자형성을 위한 Biotin 要求性에 관하여)

  • Lee, Oh-Hyoung
    • Korean Journal of Microbiology
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    • v.22 no.3
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    • pp.135-142
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    • 1984
  • The effect of biotin on the growth and sporulation of Bacillus subtilis SNU816 was investigated. When B. subtilis SNU816 was cultured on glucose as a sole carbon source, the growth was retarded markedly and usually ceased at early log phawe. But by addition of biotin to this medium, normal, rapid growth was restored. The growth rate was increased proportionally according to the concentration of exogenous biotin until it reached to 0.05㎍/ml, at which about three fold rapid growth was achieved. Also biotin was required for optimum sporulation for it facilitated the complete utilization of both glucose(Glc) and glutamic acid(Glu). Without biotin in Glc+Glu medium, about 40% of glutamic acid was remained unutilized. The dipicolinic acid content of cells cultured in Glc+Glu medium without biotin was markedly small and sporulation was suppressed before free spore release. Since biotin could be partiallyreplaced by one of TCA cycle intermediates such as oxalacetic acid, citric acid, or glutamic acid in enhancing growth in Glc medium, it was postulated that this strain might have a defect in converting pyruvate to oxalacetate which process is known to be mediated by pyruvate carboxylase that requires biotin as a cofactor.

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The molecular mechanism of propionate-regulating gluconeogenesis in bovine hepatocytes

  • Rui Pang;Xiao Xiao;Tiantian Mao;Jiajia Yu;Li Huang;Wei Xu;Yu Li;Wen Zhu
    • Animal Bioscience
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    • v.36 no.11
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    • pp.1693-1699
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    • 2023
  • Objective: Cows that are nursing get around 80% of their glucose from liver gluconeogenesis. Propionate, a significant precursor of liver gluconeogenesis, can regulate the key genes involved in hepatic gluconeogenesis expression, but its precise effects on the activity of enzymes have not yet been fully elucidated. Therefore, the aim of this study was to investigate the effects of propionate on the activity, gene expression, and protein abundance of the key enzymes involved in the gluconeogenesis of dairy cow hepatocytes. Methods: The hepatocytes were cultured and treated with various concentrations of sodium propionate (0, 1.25, 2.50, 3.75, and 5.00 mM) for 12 h. Glucose content in the culture media was determined by an enzymatic coloring method. The activities of gluconeogenesis related enzymes were determined by enzyme linked immunosorbent assay kits, and the levels of gene expression and protein abundance of the enzymes were detected by real-time quantitative polymerase chain reaction and Western blot, respectively. Results: Propionate supplementation considerably increased the amount of glucose in the culture medium compared to the control (p<0.05); while there was no discernible difference among the various treatment concentrations (p>0.05). The activities of cytoplasmic phosphoenolpyruvate carboxylase (PEPCK1), mitochondrial phosphoenolpyruvate carboxylase (PEPCK2), pyruvate carboxylase (PC), and glucose-6-phosphatase (G6PC) were increased with the addition of 2.50 and 3.75 mM propionate; the gene expressions and protein abundances of PEPCK1, PEPCK2, PC, and G6PC were increased by 3.75 mM propionate addition. Conclusion: Propionate encouraged glucose synthesis in bovine hepatocytes, and 3.75 mM propionate directly increased the activities, gene expressions and protein abundances of PC, PEPCK1, PEPCK2, and G6PC in bovine hepatocytes, providing a theoretical basis of propionate-regulating gluconeogenesis in bovine hepatocytes.

A Study on the Source of Reductants for Nitrate Reduction in Rice (Oryza sativa cv. Tongil) Roots (벼(Oryza sativa cv. Tongil) 뿌리에 있어서 Nitrate 환원에 필요한 환원력의 공급원에 관한 연구)

  • Chang, Nam-Kee;Choe, Hong-Gwan
    • The Korean Journal of Ecology
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    • v.7 no.2
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    • pp.85-90
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    • 1983
  • There was a decrease in nitrate reductase activity (NRA) measured in vivo in rice roots (Oryza sativa cv. Tongil) grown in anaerobic culture solution. But it was reversed by addition of malonate to the in vivo nitrate reduction assay medium. Malonate increased the in vivo NRA during 2-5 hours incubation and decreased it in longer incubation hours. In vivo NRA was stimulated by addition of NaHCO3 to the assay medium, but not by Na2CO3. The stimulation of NRA by NaHCO3 was not observed in shoot removed rice roots. It is suggested that CO2 from NaHCO3 is carboxylated by phosphoenol pyruvate carboxylase, results in increasing the malate contents in the roots, and stimulates the in vivo NRA. NADH needed in nitrate reduction is supported by malate oxidation. In rice roots, it seems probable that malate oxidation in the mitochondria is more important to nitrate reduction than malae oxidation in cytoplasm.

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Organic Acidopathies as Etiologic Diseases of Seizure Disorders in Korean Childhood and Adolescent Age Group (한국인 소아청소년기 발작의 원인질환으로서의 유기산대사이상질환)

  • Kim, Hui Kwon;Lee, Jong Yoon;Lee, Ye Seung;Bae, Eun Joo;Oh, Phil Soo;Park, Won Il;Lee, Hong Jin
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.12 no.1
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    • pp.23-34
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    • 2012
  • Purpose: Acute symptomatic seizures are caused by structural changes, inflammation or metabolic changes of brain, such as tumor, stroke, meningitis, encephalitis and metabolic disorders. Inherited metabolic disorders that can cause seizures are organic acidopathies, lysosomal storage disorders, peroxisomal disorders and mineral disorders. We have done this study to find out the importance of organic acidopathies causing seizure disorders in Korean childhood and adolescent patients. Method: Retrograde analysis for 1,306 patients with seizure disorders whose clinical informations are available and have done urine organic acid analysis for 5 years period, between Jan. 1st 2007 to Dec. 31th 2011. Statistical analysis was done with Student's t test using SPSS. Result: Out of 1,306 patients, 665 patients (51%) showed abnormalities on urine organic acid analysis. The most frequent disease was mitochondrial respiratory chain disorders (394, 30.1%), followed by mandelic aciduria (127, 9.7%), ketolytic defects (81, 6.2%), 3-hydroxyisobutyric aciduria (19, 1.4%), glutaric aciduria type II (10, 0.8%), ethylmalonic aciduria (4), propionic aciduria (4), methylmalonic aciduria (3), glutaric aciduria type I (3), pyruvate dehydrogenase deficiency (3), pyruvate carboxylase deficiency (3), isovaleric aciduria (2), HMG-CoA lyase deficiency (2), 3-methylcrotonylglycinuria (2), fatty acid oxidation disorders (2), fumaric aciduria (1), citrullinemia (1), CPS deficiency (1), MCAD deficiency (1). Conclusion: On neonatal period, mandelic aciduria due to infection was found relatively frequently. Mitochondrial disorders are most frequent etiologic disease on all age group, followed by ketolytic defects and various organic acidopathies. The number and diversities of organic acidopathies emphasize meticulous evaluation of basic routine laboratory examinations and organic acid analysis with initial sample on every seizure patient.

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Effect of Increased Glutamate Availability on L-Ornithine Production in Corynebacterium glutamicum

  • Hwang, Joong-Hee;Hwang, Gui-Hye;Cho, Jae-Yong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.704-710
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    • 2008
  • Glutamate availability in the argF-argR-proB${\Delta}$ strain of Corynebacterium glutamicum was increased by addition of glutamate to the cell or inactivation of the phosphoenolpyruvate carboxykinase activity and simultaneous overexpression of the pyruvate carboxylase activity to assess its effect on L-ornithine production. When glutamate was increased in an L-ornithine-producing strain, the production of L-ornithine was not changed. This unexpected result indicated that the intracellular concentration and supply of glutamate is not a rate-limiting step for the L-ornithine production in an L-ornithine-producing strain of C. glutamicum. In contrast, overexpression of the L-ornithine biosynthesis genes (argCJBD) resulted in approximately 30% increase of L-ornithine production, from 12.73 to 16.49 mg/g (dry cell weight). These results implied that downstream reactions converting glutamate to L-ornithine, but not the availability of glutamate, is the rate-limiting step for elevating L-ornithine production in the argF-argR-proB${\Delta}$ strain of C. glutamicum.

Folic acid supplementation prevents high fructose-induced non-alcoholic fatty liver disease by activating the AMPK and LKB1 signaling pathways

  • Kim, Hyewon;Min, Hyesun
    • Nutrition Research and Practice
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    • v.14 no.4
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    • pp.309-321
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    • 2020
  • BACKGROUND/OBJECTIVES: The present study aimed to evaluate the effects of folic acid supplementation in high-fructose-induced hepatic steatosis and clarify the underlying mechanism of folic acid supplementation. MATERIALS/METHODS: Male SD rats were fed control, 64% high-fructose diet, or 64% high-fructose diet with folic acid for eight weeks. Plasma glutamate-pyruvate transaminase, glutamate-oxaloacetate transaminase, lipid profiles, hepatic lipid content, S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH) were measured. RESULTS: The HF diet significantly increased hepatic total lipid and triglyceride (TG) and decreased hepatic SAM, SAH, and SAM:SAH ratio. In rats fed a high fructose diet, folic acid supplementation significantly reduced hepatic TG, increased hepatic SAM, and alleviated hepatic steatosis. Moreover, folic acid supplementation in rats fed high fructose enhanced the levels of phosphorylated AMP-activated protein kinase (AMPK) and liver kinase B (LKB1) and inhibited phosphorylation of acetyl coenzyme A carboxylase (ACC) in the liver. CONCLUSIONS: These results suggest that the protective effect of folic acid supplementation in rats fed high fructose may include the activation of LKB1/AMPK/ACC and increased SAM in the liver, which inhibit hepatic lipogenesis, thus ameliorating hepatic steatosis. The present study may provide evidence for the beneficial effects of folic acid supplementation in the treatment of non-alcoholic fatty liver disease.

Limiting Concentrate during Growing Period Affect Performance and Gene Expression of Hepatic Gluconeogenic Enzymes and Visfatin in Korean Native Beef Calves

  • Chang, S.S.;Lohakare, J.D.;Singh, N.K.;Kwon, E.G.;Nejad, J.G.;Sung, K.I.;Hong, S.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.2
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    • pp.202-210
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    • 2013
  • This study elucidated the effects of limited concentrate feeding on growth, plasma profile, and gene expression of gluconeogenic enzymes and visfatin in the liver of Hanwoo beef calves. The purpose of this study was to test that reducing the amount of concentrate would partially be compensated by increasing the intake of forage and by altering the metabolic status. The study utilized 20 Korean native beef calves (Hanwoo; 60 to 70 d of age) divided into two groups of 10 calves each for 158 d. Control group calves received the amount of concentrate as per the established Korean feeding standards for Hanwoo, whereas calves in the restricted group only received half the amount of concentrate as per standard requirements. Good quality forage (Timothy hay) was available for ad libitum consumption to both groups. Since calves were with their dam until 4 months of age in breeding pens before weaning, the intake of milk before weaning was not recorded, however, the concentrate and forage intakes were recorded daily. Body weights (BW) were recorded at start and on 10 d interval. Blood samples were collected at start and at 50 d interval. On the final day of the experiment, liver biopsies were collected from all animals in each group. The BW was not different between the groups at all times, but tended to be higher (p = 0.061) only at final BW in control than restricted group. Total BW gain in the control group was 116.2 kg as opposed to 84.1 kg in restricted group that led to average BW gain of 736 g/d and 532 g/d in respective groups, and the differences were significant (p<0.01). As planned, the calves in the control group had higher concentrate and lower forage intake than the restricted group. The plasma variables like total protein and urea were higher (p<0.05) in control than restricted group. The mRNA expressions for the gluconeogenic enzymes such as cytosolic phosphoenol pyruvate carboxykinase (EC 4.1.1.32) and pyruvate carboxylase (EC 6.4.1.1), and visfatin measured by quantitative real-time PCR in liver biopsies showed higher expression (p<0.05) in restricted group than control. Overall, restricting concentrate severely reduced the growth intensity and affected few plasma indices, and gene expression in liver was increased indicating that restricting concentrate in the feeding schemes during early growth for beef calves is not advocated.

NMR-based metabolomic profiling of the liver, serum, and urine of piglets treated with deoxynivalenol

  • Jeong, Jin Young;Kim, Min Seok;Jung, Hyun Jung;Kim, Min Ji;Lee, Hyun Jeong;Lee, Sung Dae
    • Korean Journal of Agricultural Science
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    • v.45 no.3
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    • pp.455-461
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    • 2018
  • Deoxynivalenol (DON), a Fusarium mycotoxin, causes health hazards for both humans and livestock. Therefore, the aim of this study was to investigate the metabolic profiles of the liver, serum, and urine of piglets fed DON using proton nuclear magnetic resonance ($^1H-NMR$) spectroscopy. The $^1H-NMR$ spectra of the liver, serum, and urine samples of the piglets provided with feed containing 8 mg DON/kg for 4 weeks were aligned and identified using the icoshift algorithm of MATLAB $R^2013b$. The data were analyzed by multivariate analysis and by MetaboAnalyst 4.0. The DON-treated groups exhibited discriminating metabolites in the three different sample types. Metabolic profiling by $^1H-NMR$ spectroscopy revealed potential metabolites including lactate, glucose, taurine, alanine, glycine, glutamate, creatine, and glutamine upon mycotoxin exposure (variable importance in the projection, VIP > 1). Forty-six metabolites selected from the principal component analysis (PCA) helped to predict sixty-five pathways in the DON-treated piglets using metabolite sets containing at least two compounds. The DON treatment catalyzed the citrate synthase reactions which led to an increase in the acetate and a decrease in the glucose concentrations. Therefore, our findings suggest that glyceraldehyde-3-phosphate dehydrogenase, citrate synthase, ATP synthase, and pyruvate carboxylase should be considered important in piglets fed DON contaminated feed. Metabolomics analysis could be a powerful method for the discovery of novel indicators underlying mycotoxin treatments.