• Title/Summary/Keyword: amino acid metabolism

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Transcriptome Analysis of Phosphate Starvation Response in Escherichia coli

  • Baek, Jong-Hwan;Lee, Sang-Yup
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.244-252
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    • 2007
  • Escherichia coli has a PhoR-PhoB two-component regulatory system to detect and respond to the changes of environmental phosphate concentration. For the E. coli W3110 strain growing under phosphate-limiting condition, the changes of global gene expression levels were investigated by using DNA microarray analysis. The expression levels of some genes that are involved in phosphate metabolism were increased as phosphate became limited, whereas those of the genes involved in ribosomal protein or amino acid metabolism were decreased, owing to the stationary phase response. The upregulated genes could be divided into temporarily and permanently inducible genes by phosphate starvation. At the peak point showing the highest expression levels of the phoB and phoR genes under phosphate-limiting condition, the phoB- and/or phoR-dependent regulatory mechanisms were investigated in detail by comparing the gene expression levels among the wild-type and phoB and/or phoR mutant strains. Overall, the phoB mutation was epistatic over the phoR mutation. It was found that PhoBR and PhoB were responsible for the upregulation of the phosphonate or glycerol phosphate metabolism and high-affinity phosphate transport system, respectively. These results show the complex regulation by the PhoR-PhoB two-component regulatory system in E. coli.

Effects of amino acids on ethanol metabolism and oxidative stress in the ethanol-perfused rat liver

  • Park, Yeong-Chul;Oh, Se-In;Lee, Mee-Sook;Park, Sang-Chul
    • Environmental Mutagens and Carcinogens
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    • v.16 no.1
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    • pp.13-18
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    • 1996
  • One mechanism of free-radical production by ethanol is suggested to be through the intracellular conversion of XDH to XO by increased ratio of NADH to NAD. The major mechanism for physiological compensation of cytosolic NADH/NAD balance is the malate/aspartate shutfie. Therefore, it is important to develop the method to improve the efficiency of malate/aspartate shuttle in ethanol metabolism. In the present study, various amino acids and organic acid involved in the shuttle were tested for their functional efficiency in modulating shuttle in the ethanol-perfused rat liver. The rate of ethanol oxidation in the liver perfused with aspartate alone or aspartate in combination with pyruvate, respectively, was increased by about 10% compared to control liver, but not in the tissues perfused with glummate, cysteine or pyruvate alone. Though glummate, cysteine and pyravate did not affect the ethanol oxidation significanfiy, they showed some suppresive effect on the ethanol-induced radical generation monitored by protein carbonylation analysis. Among the tested components, aspartate is confirmed to be the most efficient as a metabolic regulator for both ethanol oxidation and ethanol-induced oxidative stress in our perfusion system. These effects of aspartate would result from NAD recycling by its supplementation through the coupled aspartate aminotransferase/malate dehydrogenase reactions and the malate-aspartate shuttle.

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Subcloning and DNA Sequencing of the Phenol Regulatory Genes in Ralstonia eutropha JMP134 (Ralstonia eutropha JMP134에서 페놀분해에 관여하는 조절유전자의 Subcloning 및 염기서열 분석)

  • ;Subramanian Chitra
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.260-266
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    • 2002
  • In this study, chromosomal DNA fragment related to the regulation of phenol metabolism in Ralstonia eutropha JMP 134 was cloned and sequenced. The result has shown that two open reading frames (ORF1 and ORF2) exist on this regulatory region. ORF1, which initiates from 454 bp downstream of the stop codon of the phenol hydroxylase genes, was found to be composed of 501 amino acids. ORF2, whose start codon is overlapped with the stop codon of ORFl, was found to contain 232 amino acids. The comparison of amino acid sequences with other proteins has revealed that ORF1 belongs to the family of NtrC transcriptional activator, whereas ORF2 shares high homology with the family of GntR protein, which is known to be a negative regulator. ORF1 and ORF2 were designated as a putative positive regulator, phlR2 and a negative regulator phlA, respectively. Possible regulatory mechanisms of phenol metabolism in this strain was discussed.

An investigation on fermentative profile, microbial numbers, bacterial community diversity and their predicted metabolic characteristics of Sudangrass (Sorghum sudanense Stapf.) silages

  • Wang, Siran;Li, Junfeng;Zhao, Jie;Dong, Zhihao;Shao, Tao
    • Animal Bioscience
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    • v.35 no.8
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    • pp.1162-1173
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    • 2022
  • Objective: This study aimed to investigate the fermentation profiles, bacterial community and predicted metabolic characteristics of Sudangrass (Sorghum sudanense Stapf.) during ensiling. Methods: First-cutting Sudangrass was harvested at the vegetative stage and ensiled in laboratory-scale silos (1 L capacity). Triplicate silos were sampled after 1, 3, 7, 15, 30, and 60 days of ensiling, respectively. The bacterial communities on day 3 and 60 were assessed through high-throughput sequencing technology, and 16S rRNA-gene predicted functional profiles were analyzed according to the Kyoto encyclopedia of genes and genomes using Tax4Fun. Results: The Sudangrass silages showed good fermentation quality, indicated by higher lactic acid contents, and lower pH, butyric acid and ammonia nitrogen contents. The dominant genus Lactococcus on day 3 was replaced by Lactobacillus on day 60. The metabolism of amino acid, energy, cofactors and vitamins was restricted, and metabolism of nucleotide and carbohydrate was promoted after ensiling. The 1-phosphofructokinase and pyruvate kinase of bacterial community seemed to play important roles in stimulating the lactic acid fermentation, and the promotion of arginine deiminase could help lactic acid bacteria to tolerate the acidic environment. Conclusion: High-throughput sequencing technology combined with 16S rRNA gene-predicted functional analyses revealed the differences during the early and late stages of Sudangrass ensiling not only for distinct bacterial community but also for specific functional metabolites. The results could provide a comprehensive insight into bacterial community and metabolic characteristics to further improve the silage quality.

18-year Follow-up of Extended Newborn Screening for Metabolic and Endocrine Disorders (대사 및 내분비 질환에 대한 광범위 신생아 선별 검사의 18년 추적 관찰)

  • Song, Wung Joo;Lee, Sunho;Jeon, Young Mi;Kim, Sook Za;Jang, Mea Young
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.18 no.2
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    • pp.35-42
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    • 2018
  • Purpose: To follow up Korean patients with metabolic and endocrine disorders ascertained by Korea Genetics Research Center, and assess the long-term effectiveness of extended newborn screening program in Korea. Methods: From January 2000 to December 2017, tandem mass spectrometry and fluoroimmunoassay were employed in extended newborn screening (NBS). The NBS program obtained dried blood spots from 283,626 babies, 48 hours after birth, and screened for galactosemia, congenital hypothyroidism (CH), congenital adrenal hyperplasia (CAH), and 50 preventable inborn errors of amino acid, fatty acid, and organic acid metabolism. Results: 28 cases of amino acid disorders, 75 cases of organic acid disorders, 27 cases of fatty acid disorders, 51 cases of urea cycle disorders, 127 cases of CH, 14 cases of CAH, and 15 cases of galactosemia were ascertained through NBS and subsequent confirmatory laboratory tests. Patients with amino acid metabolic disorders, galactosemia, CH, or CAH were more likely to have a better long-term outcome if detected early. Early management of MSUD led to much better outcome in over 90%. Despite early intervention, 32% of other organic acidemia cases still resulted in developmental delay and neurological problems. Fatty acid disorders showed varied results; those with EMA and MCAD had a good outcome, but those with VLCAD had serious neurological problems and considerably higher mortality. 75% with UCD experienced serious neurological complications and higher mortality. Conclusion: The nation-wide NBS program must be accompanied by comprehensive long-term management and physician and family education of inborn errors of metabolism for a better outcome.

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Nutritional Regulation of GLUT Expression, Glucose Metabolism, and Intramuscular Fat Content in Porcine Muscle

  • Katsumata, M.;Kaji, Y.;Takada, R.;Dauncey, M.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.8
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    • pp.1297-1304
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    • 2007
  • We conducted a series of investigations in order to elucidate role of nutritional status in regulating GLUT expression and energy metabolism in porcine muscle. Firstly, the role of mild undernutrition in regulating muscle GLUT gene expression and function was studied in growing pigs (3 wk of age) on a high (H) or low (L) food intake (H = 2L) at $35^{\circ}C$ or $26^{\circ}C$. Low food intake selectively upregulates GLUT1 and GLUT4 gene expression; mRNA levels were elevated in longissimus dorsi (L. dorsi) and rhomboideus muscles but not in diaphragm or cardiac muscles. Our next step was to determine whether dietary lysine, a major primary limiting amino acid in diets for pigs, affects muscle GLUT4 expression. Pigs of 6 wk of age were pair-fed a control or low lysine (LL) diet. The control diet contained optimal amounts of all essential amino acids, including 1.15% lysine. The LL diet was similar but contained only 0.70% lysine. GLUT4 mRNA expression was upregulated by the LL diet in L. dorsi and rhomboideus muscles, whereas that in cardiac muscle was unaffected. GLUT4 protein abundance was also higher in rhomboideus muscle of animals on the LL diet. We conducted another investigation in order to elucidate effects of the LL diet on post-GLUT4 glucose metabolism. Activity of hexokinase was unaffected by dietary lysine levels while that of citrate synthase was higher both in L. dorsi and rhomboideus muscles of pigs fed on the LL diet. Glucose 6-phosphate content was higher in L. dorsi msucle in the LL group. Glycogen content was higher both in L. dorsi and rhomboideus muscles in the LL group. Further, we determined the effects of dietary lysine levels on accumulation of intramuscular fat (IMF) in L. dorsi muscle of finishing pigs. A low lysine diet (lysine content was 0.40%) meeting approximately 70% of the requirement of lysine was given to finishing pigs for two months. IMF contents in L. dorsi of the pigs given the low lysine diet were twice higher than those of the pigs fed on a control diet (lysine content was 0.65%). Finally, we proved that a well known effect of breadcrumbs feeding to enhance IMF of finishing pigs could be attributed to shortage of amino acids in diets including breadcrumbs.

Effects of Salmonella typhymurium Lipopolysaccharide Challenge on the Performance, Immune Responses and Zinc Metabolism of Laying Hens Supplemented with Two Zinc Sources

  • Cheng, Tingshui;Guo, Yuming
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.12
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    • pp.1717-1724
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    • 2004
  • The study was conducted to determine the effect of Salmonella typhymurium lipopolysaccharide (LPS) challenge on egg-laying performance, inflammatory response, zinc metabolism in layer fed diets supplemented with organic or inorganic zinc since 3-wk-old. The three dietary treatments were corn-soybean meal basal diet without supplemental zinc or with supplemental zinc at 60 mg/kg zinc from $ZnSO_4$ or zinc amino acid complex (ZnAA). At the age of 58 wk-old, twelve hens from each dietary treatment were allotted into two sub-groups. On day 1, 3, 5, 7 of the $58^{th}$ week of age, six birds of one sub-group were injected intraperitoneally (i.p.) with 2 ml LPS (1.0 $\ell$/ml) or sterile saline. Neither zinc source ${\times}$ immune challenge interaction nor zinc source effect on egg production performance was observed (p>0.05), LPS-challenge decreased egg production (p<0.04) and increased percentage of cracked eggs (p <0.01). With LPS challenged, the fever response of hens fed ZnAA peaked and subsided earlier than in hens fed $ZnSO_4$ or basal diet. Serum IL-1$\beta$ at 3 h was higher (p<0.01), but lower (p<0.001) at 12 h post-challenge with LPS in hens fed ZnAA than $ZnSO_4$. In salinetreated groups, serum IL 1$\beta$ was higher in hens fed ZnAA than the basal diet at 3 h post-injection (p<0.01). LPS-challenged birds had lower serum zinc and higher zinc sequestered in liver and spleen (p<0.001). In saline-treated birds, there was no difference in zinc concentration of serum, liver and spleen among different dietary treatments (p>0.05). Supplementation of 60 mg/kg zinc from either ZnAA or $ZnSO_4$ significantly (p<0.05) elevated metallothionein (MT) concentration in liver and spleen. MT concentration in liver of birds fed ZnAA diet was higher than in those fed $ZnSO_4$ diet (p<0.05). The magnitude of increase of hepatic and splenic MT due to LPS challenge was higher by supplementation of ZnAA than $ZnSO_4$. The results suggest that zinc amino acid complex enhanceed MT synthesis and zinc sequestered in liver and spleen and increased the sensitivity to immune response due to LPS challenge.

Effects of the Soybean Powder with Rich Aglycone Isoflavone on Lipid Metabolism and Antioxidative Activities in Hyperlipidemic Rats (고지혈증 흰쥐에서 비배당체 이소플라본 고함유 대두분말의 혈청 지질 대사 영향과 항산화효과)

  • Lim, Ae-Kyoung;Jung, Hee-Kyoung;Hong, Joo-Heon;Oh, Jung-Suk;Kwak, Jung-Hoon;Kim, Yong-Hae;Kim, Dae-Ik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.3
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    • pp.302-308
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    • 2008
  • This study was performed to evaluate the effects of soybean powder with rich aglycone isoflavone through bio-transformation on lipid metabolism and antioxidative activites in diet induced hyperlipidemic rats. The rats were randomly divided into four groups: NO (basal diet normal group), CO (high fat diet control group), BFF10 (soybean powder with rich aglycone isoflavone 10% group), and BFF20 (soybean powder with rich aglycone isoflavone 20% group). After 7 weeks of BFF10 or BFF20 diets consumption, the concentrations in serum triglyceride, total cholesterol, LDL-cholesterol, and atherogenic index ratios were significantly decreased in the BFF10 and BFF20 diets groups compared with those in the CO group. The activities of alanine amino transferase and aspartate amino transferase were significantly decreased in the BFF10 and BFF20 than those in the CO group. Thiobarbituric acid reactive substance levels of serum were decreased in BFF10 and BFF20 groups compared to that of the CO group. The super oxide dismutase activites were increased in BFF10 and BFF20 groups compared to that of the CO group.

Preparation and Availability Analysis of Collagen Peptides Obtained in Fish Scale (어류비늘에서 추출한 콜라겐펩타이드의 제조 및 유효성 분석)

  • Lee, Mi-Jin;Jeong, Noh-Hee
    • Journal of the Korean Applied Science and Technology
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    • v.26 no.4
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    • pp.457-466
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    • 2009
  • This study is manufacturing method and analysis of feasibility about collagen peptide from fish scale. This is processed by enzyme hydrolysis, isolating and refining etc. The results of analysis of nutritional composition showed protein content of collagen peptide. In the analysis of constitutive amino acids, the ratio of contents of hydroxyproline and glycine, the characteristics of collagen peptides appeared similar and the contents of glutamic acid and aspartic acid which are involved in protein metabolism. As a result of measurement of total polyphenol content and total flavonoid, it showed that collagen peptide had more contents generally, and the effect of bioactivity of pig-skin collagen peptide appeared higher although different kinds of scale collagen peptide showed a little DPPH radical scavenging ability, total antioxidant capacity by ABTS, ACE inhibitory.

Studies on the Free Amino Acids Metabolism in Germinating Mung Bean by Paper Chromatography. (Part $1{\sim}2$) (Part 2) Unknown Nitrogen Compound in Germinating Mung Bean (페파크로마토그라피에 依한 發芽綠豆의 遊離아미노酸代謝의 硏究 (第 1 報${\sim}$第 2 報) (第 2 報) 發芽綠豆中의 未知窒素化合物에 關하여)

  • Kim, Tae-Rin;Song, Chang-Won
    • Journal of the Korean Chemical Society
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    • v.5 no.1
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    • pp.15-17
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    • 1961
  • A ninhydrin positive substance(formed during germination of Mung Bean) which on a 2-dimensional chromatography(BuOH-HAc, and$ PhOH-NH_2$ gave a spot above glutamic acid was isolated by cutting out the appropriate spot on the paper and extracting the paper with water. Hydrolysis of this substance with $2N H_2SO_4\;or\;3N\;HCl\;at\;120^{\circ}C$ for 15 hours gave the spot of mainly glutamic acid, alanine and very faint of glycine and cysteine on a paper chromatography. It is suggested that the reaction of ${\gamma}$-glutamyl-cysteinecyl-glycine + L-alanine in the presence of ${\gamma}$-glutamyle transferase ${\rightarrow}$ ${\gamma}$-glutamyl-alanine + cysteinyl-glycine, takes place in germinated Mung Bean.

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