• Title/Summary/Keyword: coenzymes

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Application of Poly (Ethylene Glycol)-Bound NAD in Model Enzyme Reactor

  • Urabe, Itaru
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.510.1-510
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    • 1986
  • Many enzymes require the participation of readily dissociable coenzymes as NAD for thir catalytic activities. The continuous utilization of the enzymes requires the retention and regeneration of the coenzymes. For this purpose, several kinds of macromolecular NAD derivatives have been prepared by covalently attaching NAD to watersoluble polymers. We have prepared poly (ethylene glycol)-bound NAD (PEG-NAD) by coupling N$\^$6/-(2-carboxyethyl)-NAD to one terminal of ${\gamma}$ $\omega$-diaminoly (ethylene glycol) (Mr 3000) with water-soluble carbodiimide. PED-NAD thus obtained has one NAD moiety located at a terminal of the linear, flexible and hydrophilic chain of poly (ethylene glycol). PED-NAD has good coenzyme activity for various dehydrogenases and is applicable in a continuous enzyme reactor. To use these macromolecular NAD derivatives in an enzyme reactor, it si necessary to understand the behavior of the system in which the reactions of dehydrogenases are coupled by the recycling of the NAD derivative. We investigated the kinetic properties of a continuous enzyme reactor containing lactate dehydrogenase, alcohol dehydrogenase and PEG-NAD. The steady-state behavior of the enzyme reactor is explained by a simple kinetic model.

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Monitoring of itaconic acid production by a 2-dimensional fluorescence sensor

  • Jeong, Sang-Yun;Lee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.270-273
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    • 2001
  • The fluorescence sensor is utilized to monitor the complex fluorescence patterns of intra- and extracellular components in cultivation processes. Especially biogenic fluorophores such as proteins and peptides (tryptophan, phenylalanine), coenzymes (FAD, NAD(P)H) and vitamins (riboflavin, pyridoxine) within cells are detected by a fluorescence sensor. In this work a 2-dimensional fluorescence sensor has been used to monitor a production process of itaconic acid by Aspergillus terreus and the on-line monitored spectra data can be con-elated to off-line data measured by a few methods.

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RECOMMENDED DIETARY ALLOWANCES FOR GENOMIC STABILITY

  • Fenech, Michael
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.10a
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    • pp.9-9
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    • 2001
  • Several micronutrients (vitamins and minerals) are required as co-factors in DNA synthesis, DNA repair, DNA methylation and apoptosis. Some notable examples include (a) folic acid and vitamin B12 required for maintenance methylation of DNA and the synthesis of dTTP from dUTP, thus prevent the misincorporation of uracil into DNA, a highly mutagenic and chromosome-breaking event, (b) niacin, is essential in the form of the coenzymes NAD and NADP which act as a substrate for polyADPribose polymerase (PARP), an enzyme thought to facilitate efficient DNA repair and telomere length regulation and (c) zinc, apart from its antioxidant role as a co-factor in Cu/Zn SOD, it is required in its stabilizing role of the DNA-binding domain of p53 (residues 102-292) and thus is essential for apoptotic response to DNA damage. (omitted)

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Effects of Dietary Folate Intake on Plasma and Tissue Folate Concentrations in Rats (식이 엽산수준이 흰쥐의 혈장과 조직의 엽산함량에 미치는 영향)

  • 장남수
    • Journal of Nutrition and Health
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    • v.31 no.3
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    • pp.271-278
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    • 1998
  • Folate coenzymes are involved in one-carbon transfer reactions needed for the synthesis of nucleic acids, amino acids, and proteins which are very important for cell proliferation and differentiation. To investigate the effects of dietary folate content on plasma and tissue folate concentrations and on folate excretions in urine and feces, male Sprague-Dawley rats were raised for 4-10 weeks on semi-purified experimental diets containing 0mg, 2 mg, 8mg folate/kg diet. Folate concentrations were determined microbiologically using Lactobacillius casei (ATCC 7469). When compared to the folate adequate diet, the folate deficient diet decreased folate levels in plasma, liver and kidney , and the values were further decreased with experimental period. In rats reviving folate supplemented diets, plasma , liver and kidney folate adequate or supplemented diets, folate concentrations weer increased compared to animals on the folate adequate diet. In the folate adequate or supplemented diets, folate concentrations in the plasma and kidney were maintained at essentially the same level for 10 weeks . Folate concentrations in the liver, however, continued to increase with experimental period. Dietary folate intake seems to influence plasma and liver folate concentrations more than kidney folate concentrations. Folate excretions unrine and feces were significantly increased with dietary foalte intakes and experimental period. Folate excreted via urine was consideerably greater than that via feces. These resutls indicated that the foate supplemented diet improved plasma and tissue foalte status. Whether folate supplmentation improves foalte-dependent reactions remains to be researched.

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Biochemical Assessment of Vitamin $B_{1},\;B_{2}$ and $B_{6}$ Nutriture by Coenzyme Activation on Erythrocyte Enzymes (적혈구(赤血球) 효소활성화(酵素活性化)에 의(依)한 비타민 $B_{1}\;B_{2}$$B_{6}$ 영양상태(營養狀態)의 생화학적(生化學的) 평가(評價))

  • Tchai, Bum-Suk
    • Journal of Nutrition and Health
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    • v.10 no.4
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    • pp.24-32
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    • 1977
  • It was attempted in this study to assess the vitamin $B_{1},\;B_{2}$, and $B_6$ status in tissue by determination of erythrocyte transketolase (TK), glutathione reductase (GR), and aspartate aminotransferase (AST) activities, and their activation by their respective coenzymes, thiamine pyrophosphate, flavin-adenine dinucleotide, and pyridoxal-5-phosphate. The activities of erythrocyte enzymes were stable for more than 30 days when erythrocyte had been stored at $-20^{\circ}C$ and affirmed that the enzyme activities were more stable in the case of deep frozen sotrage of erythrocytes rather than hemolysates. The assay procedures involving ultraviolet kinetic analysis with continuous monitoring for each of enzymes have good within-batch and between-batch precisions and will be avalable in the routine laboratories for the nutritional and clinical surveys. Activity coefficient of TK, GR, and AST was studied in healthy medical students (fifteen men and twelve women, between 21 and 30 years old) on an unrestricted diet. The mean activity coefficient of TK, GR, and AST were 1.18, 1.35, and 2.01 for men, and 1.14, 1.33, and 1.83 for women, respectively. And the upper limit of normal (mean+2SD) were 1.52, 1.69, and 2.61 for men, and 1.50, 1.61, and 2.37 for women, respectively.

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New Nutritional Concepts of Vitamins and Minerals (비타민과 무기질의 새로운 영양학적 의미)

  • Youn, Hee-Shang
    • Clinical and Experimental Pediatrics
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    • v.48 no.12
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    • pp.1295-1309
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    • 2005
  • Nowadays, the nutritional deficits are rarely seen in Korea. However, an increased availability of the highly palatable energy dense, nutrient-poor foods increases the risks of obesity and deficits of vitamins and minerals in the general population. Also, optimum intake of vitamins and minerals, which varies with age and genetic back ground, might not suffice the poor, young, obese, and elderly people. Young girls and individuals participating in weight reductions and aesthetic components are prone to micronutrient deficiencies because they restrict food intake and specific micronutrient rich foods. An inadequate intake of vitamins or minerals is associated with reduced physical performance and exercise capacity, increased obesity, decreased cognitive function, increased DNA damages such as single- and double-stranded breaks or oxidative DNA lesions, and accelerated aging process and increased neuronal damages with mitochondrial oxidative decay. Most of these deleterious effects of the deficit could be prevented by a one tablet of multivitamins with a good balanced diet. High dose B vitamins are frequently administered to overcome the metabolic inadequacy to the people with the less functional enzymes with increased Km values for their coenzymes due to the single gene mutation or due to the single nucleotide polymorphisms. And some certain antioxidant vitamins are also used in large quantities to overcome the oxidative stress and to repair the damages. In this review, new nutritional concepts of some vitamins and minerals, which are widely used and useful for the children, will be discussed.

Simple Preparation of Diaphorase/Polysiloxane Viologen Polymer Modified Electrode for Sensing NAD and NADH

  • Song, Ji-Eun;Hong, Zhenyu;Nagarale, Rajaram Krishna;Shin, Woon-Sup
    • Journal of Electrochemical Science and Technology
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    • v.2 no.3
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    • pp.163-167
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    • 2011
  • Nicotinamide adenine dinucleotide, $NAD^+$, and its reduced form, NADH, play important roles as coenzymes in many enzymatic reactions. Electrochemical methods for $NAD^+$ or NADH detection or generation are drawn attention because it can provide the simple and low cost platform with fairly good sensitivity. In this study, the polysiloxane viologen polymer/diaphorase/hydrophilic polyurethane (PSV/DI/HPU) modified electrodes were simply prepared and demonstrated for bio-electrocatalytic $NAD^+$ sensors. The electrodes were co-immobilized with diaphorase and polysiloxane viologen polymer as an electron mediator followed by the overcoating with HPU membrane. The mixture of the enzyme and the electron mediator was well stabilized within HPU membrane and exhibited good reversibility and stability. The sensitivity was 0.2 $nA{\cdot}{\mu}M^{-1}$ and the detection limit was 28 ${\mu}M$ with a response time of 50 s ($t_{90%}$). The capability for NADH sensor was also observed on the PSV/DI/HPU electrode.

The Characteristics of I269S and I224S Double Mutant Horse Liver Alcohol Dehydrogenase (I269S와 I224S 이중변이 알코올 탈수소효소의 특성)

  • Ryu, Ji-Won;Lee, Kang-Man
    • YAKHAK HOEJI
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    • v.41 no.6
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    • pp.756-764
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    • 1997
  • Ile-224 in I269S mutant horse liver alcohol dehydrogenase isoenzyme S (HLADH-S) was mutated to serine by site-directed mutagenesis in order to study the role of the residue in c oenzyme binding to the enzyme. The specific activity of the I269S and I224S mutant enzyme to ethanol was increased 6-fold and all Michaelis constants($K_a,\;K_b,\;K_p,\;and\;K_q$,/TEX>) were larger than those for the wild-type and I269S enzyme. The substitution decreased the afffinity to coenzymes and increased the specific activity of the enzyme. The mutant enzyme showed the highest catalytic efficiency for octanol among the primary alcohols. But it didn`t have activities on retinoids and 5${\beta}$-cholanic acid-3-one. From these results, it was confirmed that the hydrophobic interaction of Ile-224 residue with coenzyme was related to coenzyme affinity in ADH reaction. The substitution also affected the substrate affinities to the enzyme.

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Cellular and Biochemical Alterations in L6 Myoblast Cells Induced by 6-Aminonicotinamide

  • Jang, Min-Young;Kim, Sun-Jung;Shin, Sook;Park, In-Kook
    • Animal cells and systems
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    • v.11 no.1
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    • pp.17-22
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    • 2007
  • The effects of antimetabolite 6-AN (6-amino-nicotinamide) on viability and morphology of L6 myoblast cells have been investigated. 6-AN ($100{\mu}M$) induced a time-dependent decrease in cell viability with respect to the untreated control cells. Following 6-AN administration the viability rate started to decline sharply, reaching about 23% of the untreated control cells at 48 h. Inverted phase-contrast microscopy revealed that 6-AN caused characteristic morphological changes such as irregularly elongated and stellate shape of cells, round-shaped nucleus, cytoplasmic vacuolization, irregular cell arrangements and formation of large spaces among cell clusters. The concentrations of ATP and $NAD^{+}$ in the 6-AN treated cells were significantly lower (p < 0.01) than those of the untreated control cells. In contrast, the concentration of AMP was significantly increased by the 6-AN treatment. Activities of catalase, superoxide dismutase and glutathione peroxidase in 6-AN treated cells were significantly higher (p < 0.01) than those of the untreated control cells. The activities of glyceraldehyde-3-phosphate dehydrogenase in 6-AN treated cells were significantly lower (p < 0.01) than those of the untreated control cells. The results suggest that 6-AN caused marked reduction of cell viability and alterations of some important metabolites and enzymes.

Modulation of DNA methylation by one-carbon metabolism: a milestone for healthy aging

  • Sang-Woon Choi ;Simonetta Friso
    • Nutrition Research and Practice
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    • v.17 no.4
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    • pp.597-615
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    • 2023
  • Healthy aging can be defined as an extended lifespan and health span. Nutrition has been regarded as an important factor in healthy aging, because nutrients, bioactive food components, and diets have demonstrated beneficial effects on aging hallmarks such as oxidative stress, mitochondrial function, apoptosis and autophagy, genomic stability, and immune function. Nutrition also plays a role in epigenetic regulation of gene expression, and DNA methylation is the most extensively investigated epigenetic phenomenon in aging. Interestingly, age-associated DNA methylation can be modulated by one-carbon metabolism or inhibition of DNA methyltransferases. One-carbon metabolism ultimately controls the balance between the universal methyl donor S-adenosylmethionine and the methyltransferase inhibitor S-adenosylhomocysteine. Water-soluble B-vitamins such as folate, vitamin B6, and vitamin B12 serve as coenzymes for multiple steps in one-carbon metabolism, whereas methionine, choline, betaine, and serine act as methyl donors. Thus, these one-carbon nutrients can modify age-associated DNA methylation and subsequently alter the age-associated physiologic and pathologic processes. We cannot elude aging per se but we may at least change age-associated DNA methylation, which could mitigate age-associated diseases and disorders.