• Title/Summary/Keyword: Adenosine 5'-Monophosphate

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Purification and Characterization of Adenosine deaminase from Aspergillus oryzae (Aspergillus oryzae에서 Adenosine Deaminase의 정제와 특성)

  • Choi, Hye-Seon
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.54-62
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    • 1993
  • Intracellular adenosine deaminase (ADA) from Aspergillus oryzae was purified using ammonium sulfate fractionation, a DEAE-Sephadex A-50 anion exchange chromatography, an ultrafiltration using a PM 10 membrane and two times of Sephadex G-100 gel filtration chromatography. The enzyme was purified 151 fold with a 9% recovery. Purified enzyme gave a single protein band with a molecular weight of 105,000 delton. The enzyme was reasonably stable. The enzyme activity was kept even after 1 hr incubation at 55.deg.C, but decreased significantly at 60.deg.C. The pH optimum was found to be from 6.5 to 7.5. Among tested compounds, the substrate activity was found with adenosine, adenine arainofuranoside, formymcin A, 2'-deoxyadenosine, 3'-deoxyadenosine, 2', 3'-isopropylidene adenosine, 2,6-diaminopurine deoxyriboside, .betha.-nicotinamide adenine dinucleotide (reduced form), 6-chloropurine riboside, 2'-adenine monophosphate (AMP), 3'-AMP and 5'-AMP. The values of Km of adenosine and 2'-deoxyadenosine were calculated to be 500 and .$710\mu$m, respectively. ADA was sensitivite to $Zn^{2+}$, $^Cu{2+}$ and $Fe^{3+}$, p-chloromercuribenzoate and mersalyl acid inactivated the enzyme. The activity of enzyme was not changed when ADA was incubated with dithiothreititol, 2-mercaptoethanol, N-ethylmaleimide, iodoacetic acid and iodoacetamide.

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The influence of ultrasound and adenosine 5'-monophosphate marination on tenderness and structure of myofibrillar proteins of beef

  • Zou, Ye;Yang, Heng;Zhang, Muhan;Zhang, Xinxiao;Xu, Weimin;Wang, Daoying
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.10
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    • pp.1611-1620
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    • 2019
  • Objective: The aim was to investigate the influence of ultrasound and adenosine 5'-monophosphate (AMP) marination (UAMP) on tenderness and structure of myofibrillar proteins of beef. Methods: Five groups, the untreated meat (Control), deionized water marination (DW), ultrasound followed by DW (UDW), AMP marination (AMP), and ultrasound followed by AMP (UAMP) were studied. Myofibrillar fragmentation, cooking loss, shear force, thermograms, histological observation of meats and myofibrillar proteins properties were investigated in these different treatments. Results: The results showed that UAMP significantly increased myofibrillar fragmentation index from 152 (Control), 231 (AMP), and 307 (UDW) to 355 (p<0.05), respectively. The lowest cooking loss, shear force and peak denaturation temperature were observed in UAMP. In histological observation, UDW and UAMP had more fragmented muscular bundles than the others. Furthermore, a drastic increase in ${\alpha}$-helix and decrease in ${\beta}$-sheet of myofibrillar proteins was observed in UAMP, implying the disaggregation of protein samples. The synchronous fluorescence spectra of myofibrillar proteins in UAMP suggested the combination of ultrasound and AMP could accelerate the unfolding molecular structure and destroying hydrophobic interactions. The results of circular dichroism and synchronous fluorescence spectra for myofibrillar proteins coincided with the microstructures of beef. Conclusion: The results indicate that ultrasound combined with AMP improved meat tenderness not only by disruption in muscle integrity, increasing water retention, but also altering their spatial structure of myofibrillar proteins.

Purification and Characterization of Pyrimidine Nucleotide N-Ribosidase from Pseudomonas oleovorans

  • YU, Tae-Shick
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.573-578
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    • 2005
  • Pyrimidine nucleotide N-ribosidase (pyrimidine 5'-nucleotide phosphoribo(deoxyribo)hydrolase/pyrimidine 5'-nucleotide nucleosidase, EC 3.2.2.10) catalyzes the breakdown of pyrimidine 5'-nucleotide into pyrimidine base and ribose(deoxyribo)-5-phosphate. However, detailed characteristics of the enzyme have not yet been reported. The enzyme was purified to homogeneity 327.9-fold with an overall yield of $6.1\%$ from Pseudomonas oleovorans ATCC 8062. The enzyme catalyzed cytidine monophosphate (CMP) and uridine monophosphate (UMP), but not adenosine monophosphate (AMP) and guanosine monophosphate (GMP). The enzyme optimally metabolized CMP at pH 6.0 and UMP at around 8.5, and the optimum temperature for the overall enzyme reaction was found to be $37^{\circ}C$. The $K_m$ values of the enzyme for CMP (at pH 6.0) and UMP (at pH 8.5) were 1.6 mM and 1.1 mM, respectively. AMP, deoxyCMP, and deoxyUMP were very effective inhibitors of the reaction. Double-reciprocal plots obtained in the absence and in the presence of AMP revealed that this inhibitory effect was of the mixed competitive type with respect to the breakdown of CMP and of the noncompetitive type with respect to the breakdown of UMP. In the presence of AMP, the enzyme followed sigmoid kinetics with respect to each substrate.

Anti-platelet Effects of Dimethyl Sulfoxide via Down-regulation of COX-1 and $TXA_2$ Synthase Activity in Rat Platelets

  • Ro, Ju-Ye;Lee, Hui-Jin;Ryu, Jin-Hyeob;Park, Hwa-Jin;Cho, Hyun-Jeong
    • Biomedical Science Letters
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    • v.20 no.2
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    • pp.70-76
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    • 2014
  • In this study, we investigated the effect of DMSO, a highly dipolar organic liquid, in collagen ($5{\mu}g/ml$)-stimulated platelet aggregation. DMSO inhibited platelet aggregation at 0.5% by inhibiting production of thromboxane $A_2$ ($TXA_2$) which was associated with blocking cyclooxygenase (COX)-1 activity and $TXA_2$ synthase. In addition, DMSO significantly increased the formation of cyclic adenosine monophosphate (cAMP) from adenosine triphosphate (ATP) and cyclic guanosine monophosphate (cGMP) from guanosine triphosphate (GTP). On the other hand, DMSO (0.1~0.5% concentration) did not affect the LDH release which indicates the cytotoxicity. Based on these results, DMSO has anti-platelet effect by regulation of several platelet signaling pathways, therefore we suggest that DMSO could be a novel strategy on many thrombotic disorders.

Kinetics of Hydrogen-Deuterium Exchange of 8-CH Groups in Adenosine 5$^\prime$-Monophosphate and Guanosine 5$^\prime$-Monophosphate by Laser Raman Spectroscopy

  • Kim, Sang-Kyu;Kim, Myung-Soo;Suh, Se-Won
    • Bulletin of the Korean Chemical Society
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    • v.6 no.5
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    • pp.270-272
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    • 1985
  • The rate constants of the hydrogen-deuterium exchange of 8-CH groups in 5'-rAMP and 5'-GMP were measured by laser Raman spectroscopy. The Arrhenius activation energies calculated from the rate constants measured as a function of temperature were similar for both compounds. However, the effects of pD on exchange rate constants were different for the two compounds. Our kinetic data support the exchange reaction mechanism involving an ylide type intermediate.

Studies on the Fermentative Production of Guanosine -5′-monophosphate by Microorganism (Part 1) Derivation of XMP-aminase Producing Mutants (미생물에 의한 5′-GMP의 생산에 관한 연구 (제1보) XMP-aminase 생산균주의 분리)

  • 배종찬;손충홍;공운영;유주현
    • Microbiology and Biotechnology Letters
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    • v.7 no.3
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    • pp.127-133
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    • 1979
  • By the treatment of several mutagens, a number of 5'-guanylic acid producing mutants from 5'-xan-thylic acid were obtained from Brevibacterium ammoniagenes ATCC 6871. The indispensensable genetic-characters of the mutants were adenine requirement, lack of GMP-reductase and mutation to adenosine resistance from adenosine sensitiveness. Main product from 5'-xanthylic acirl by strain BA-17-2 was 5'-guanulic acid, and was isolated in a crystalline form by the use of anion exchange resin, Duolite 102 D. The isolated crystalline was identified as 5'-guanylic acid by means of paper chromatography, ultrav-iolet absorption spectra, and infrared absorption spectrum.

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Nucleotide Analysis in Korean Dairy Products Using High- Performance Liquid Chromatography with Diode Array Detector

  • Won, Jong-Eun;Bang, Han-Yeol;Kwak, Byung-Man;Park, Jong-Su;Kim, Gui-Ran;Kwon, Joong-Ho
    • Food Science of Animal Resources
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    • v.39 no.1
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    • pp.93-101
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    • 2019
  • Nucleotides play important roles in numerous intracellular biochemical processes and are used in infant formulas and other dairy products. However, domestic analytical methods for assessing nucleotide content in products have not yet been established, and therefore, methods for determining nucleotide content are urgently required. A rapid and simple analytical method for determining the content of five types of nucleotides in dairy products was improved using solid phase extraction clean-up and high-performance liquid chromatography with diode array detector. The extraction solvent used in the AOAC method was not well dissolved and was changed to hydrophilic EDTA-Na. In addition, the results obtained using the isocratic elution method and a single wavelength were similar to those obtained using the AOAC method, and the time taken for analysis was shortened from 40 min to 25 min. The process of method validation revealed the following parameters: accuracy (84.69%-102.72%), precision (1.51%-6.82%), linearity (0.999), and limit of detection (cytidine 5'-monophosphate, 0.09 mg/L; uridine 5'-monophosphate, 0.11 mg/L; adenosine 5'-monophosphate, 0.12 mg/L; guanosine 5'-monophosphate, 0.11 mg/L; and inosine 5'-monophosphate, 0.14 mg/L). The method was also used to determine the nucleotide concentration in 25 samples (infant formulas, 1.99-29.39 mg/100 g; and cow milk, 0.28-0.83 mg/100 g). The newly improved method was appropriate for analyzing nucleotides in infant formulas and other dairy products faster when compared to conventional methods.

Development of natural Seasoning Based on Mushroom (버섯 천연조미료 제조에 관한 연구)

  • 박미자;이종숙;이별나;이재성
    • Journal of the East Asian Society of Dietary Life
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    • v.11 no.3
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    • pp.196-203
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    • 2001
  • This study was conducted to provide basic data for the development of natural seasoning. The quality chatacteristics of water exacts from Pleurotus ostreatus and Lentinus edodes were determind. Also, savory taste, fragrance, color and overall preference was evaluated. The preference test on the seasonings prepared from Pleurotus ostreatus and Lentinus edodes with the addition of sea tangle was also performed. The crude protein content of dried, Lentinus edodes and Pleurotur ostreatus was 20.4% and 30.4% respectively. The total organic acid contents were higher in Plourotus ostreatus than in Lentinus edodes extracts. The total free amino acid adn total nucleotide contents were higher in Pleurotus ostreatus than in Lentinus edodes extracts. Three nucleotide, adenosine monophosphate(AMP) , guanosine monophosphate(GMP), xanthine monophosphate(XMP) were found in mushroom seasonings while inosine monophosphate(IMP) was not detected. Pleurotus ostreatus extract showed higher score in savory taste and overall acceptability than Lentinus edodes extract, Pleurotur ostreatus seasoning added with 1% sea tangle and Lentinus edodes seasoning added with 1.5% sea tangle were rated as high in overall quality In a sensory test of foods cooked with mushroom seasoning. It was shown that panels in their twenties well used to the taste of monosodium glutamate while the people in their thirties preferred the specific flavor of the mushroom seasoning.

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Comparision of Regulatory Action of cAMP and cGMP on the Activation of Neutrophil Responses

  • Han, Chang-Hwang;Yoon, Young-Chul;Shin, Yong-Kyoo;Han, Eun-Sook;Lee, Chung-Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.1
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    • pp.97-105
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    • 1997
  • The regulatory role of cyclic nucleotides in the expression of neutrophil responses has been examined. fMLP-stimulated superoxide production in neutrophils was inhibited by dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP), histamine, adenosine + theophylline, cAMP elevating agents, and 8-bromoguanosine 3' ,5' -cyclic monophosphate (8-BrcGMP) and sodium nitroprusside, cGMP elevating agents. Staurosporine, a protein kinase C inhibitor, genistein, a protein tyrosine kinase inhibitor and chlorpromazine, a calmodulin inhibitor, inhibited superoxide production by fMLP, but they did not further affect the action of DBcAMP on the stimulatory action of fMLP. DBcAMP, histamine, adenosine+theophylline and genistein inhibited myeloperoxidease release evoked by fMLP, whereas BrcGMP, sodium nitroprusside and staurosporine did not affect it. The elevation of $[Ca^{2+}]_i$ evoked by fMLP was inhibited by genistein and chlorpromazine but was not affected by staurosporine. DBcAMP exerted little effect on the initial peak in $[Ca^{2+}]_i$ response to fMLP but effectively inhibited the sustained rise. On the other hand, BrcGMP significantly inhibited both phases. fMLP-induced $Mn^{2+}$ influx was inhibited by either DBcAMP or BrcGMP. These results suggest that fMLP-stimulated neutrophil responses may be regulated by cAMP more than cGMP. cAMP and cGMP appear not affect stimulated responses by direct protein kinase C activation. Their regulatory action on the stimulated neutrophil responses may be not influenced by other activation processes.

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Probiotics Increase Intramuscular Fat and Improve the Composition of Fatty Acids in Sunit Sheep through the Adenosine 5'-Monophosphate-Activated Protein Kinase (AMPK) Signaling Pathway

  • Yue Zhang;Duo Yao;Huan Huang;Min Zhang;Lina Sun;Lin Su;LiHua Zhao;Yueying Guo;Ye Jin
    • Food Science of Animal Resources
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    • v.43 no.5
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    • pp.805-825
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    • 2023
  • This experiment aims to investigate the impact of probiotic feed on growth performance, carcass traits, plasma lipid biochemical parameters, intramuscular fat and triglyceride content, fatty acid composition, mRNA expression levels of genes related to lipid metabolism, and the activity of the enzyme in Sunit sheep. In this experiment, 12 of 96 randomly selected Sunit sheep were assigned to receive the basic diet or the basic diet supplemented with probiotics. The results showed that supplementation with probiotics significantly increased the loin eye area, and decreased plasma triglycerides and free fatty acids, increasing the content of intramuscular fat and triglycerides in the muscle and improving the composition of the fatty acids. The inclusion of probiotics in the diet reduced the expression of adenosine 5'-monophosphate-activated protein kinase alpha 2 (AMPKα2) mRNA and carnitine palmitoyltransferase 1B (CPT1B) mRNA, while increasing the expression of acetyl-CoA carboxylase alpha (ACCα) mRNA, sterol regulatory element-binding protein-1c (SREBP-1c) mRNA, fatty acid synthase mRNA, and stearoyl-CoA desaturase 1 mRNA. The results of this study indicate that supplementation with probiotics can regulate fat deposition and improves the composition of fatty acids in Sunit sheep through the signaling pathways AMPK-ACC-CPT1B and AMPK-SREBP-1c. This regulatory mechanism leads to an increase in intramuscular fat content, a restructuring of muscle composition of the fatty acids, and an enhancement of the nutritional value of meat. These findings contribute to a better understanding of the food science of animal resources and provide valuable references for the production of meat of higher nutritional value.