• Title/Summary/Keyword: pyridoxamine

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Modification of Carboxyl Residues of Proteins with Pyridoxamine as a Fluorophore

  • Kwon, Oh-Shin
    • BMB Reports
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    • v.29 no.3
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    • pp.215-220
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    • 1996
  • A general procedure to quantitate the reaction of carbodiimides with carboxy groups of proteins is described. Pyridoxamine reacts with the o-acylisourea intermediate generated during the reaction of carboxyl residues with carbodiimides. The extent of the reaction is determined by measuring the spectroscopic properties, absorption and emission, of pyridoxyl residues covalently attached to the proteins. Resolved pig brain aspartate aminotransferase (apoenzyme), inactivated by 1-ethyl-3-(3-dimethylamino propyl) carbodiimide, reacts with $[^{3}H]pyridoxamine$. After trypsin digestion, one peptide labeled with radioactive pyridoxyl was separated by reverse phase HPLC.

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ENZYMATIC STUDIES ON VITAMIN B6 METABOLISM

  • Kim, Young-Tae
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.133-142
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    • 1993
  • Vitamin B6(pyridoxine, pyridoxamine. and pyridoxal) is a dietary requirement in relatively small quantities for growth, health, and function in animals and fish. The metabolically active B6 is pyridoxal-5-phosphate(PLP). It does function as a coenzyme in number of enzymes(PLP-dependent enzymes) in which amino acids are metabolized, including decarboxylases, aminotransferases, sulfhydrases, tryptophanase, and hydroxylases. Vitamin B6 requirement is higher for fish because fish are fed much higher protein diet than land animals. B6 is also involved in metabolism of carbohydrates and lipids and essential for the synthesis of heme and serotonin. Deficiency signs in fish develop quickly, in cluding nervous disorders, convulsions, poor swimming coordination, skin lesions, edema, exophthalmos, and tetany. The conversion of vitamin B6 to metabolically active form(PLP) is catalyzed by pyridoxal kinase and pridoxine(pyridoxamine) oxidase. In this review, we summarized in detail the enzymatic studies on vitamin B6 metabolism and about the mechanisms and properties of a PLP-dependent enzyme.

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Vitamin B6 Deficiency, Genome Instability and Cancer

  • Wu, Xia-Yu;Lu, Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5333-5338
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    • 2012
  • Vitamin B6 functions as a coenzyme in >140 enzymatic reactions involved in the metabolism of amino acids, carbohydrates, neurotransmitters, and lipids. It comprises a group of three related 3-hydroxy-2-methyl-pyrimidine derivatives: pyridoxine (PN), pyridoxal (PL), pyridoxamine (PM) and their phosphorylated derivatives [pyridoxal 5'-phosphate (PLP) and pyridoxamine 5'-phosphate (PMP)], In the folate metabolism pathway, PLP is a cofactor for the mitochondrial and cytoplasmic isozymes of serine hydroxymethyltransferase (SHMT2 and SHMT1), the P-protein of the glycine cleavage system, cystathionine ${\beta}$-synthase (CBS) and ${\gamma}$-cystathionase, and betaine hydroxymethyltransferase (BHMT), all of which contribute to homocysteine metabolism either through folate-mediated one-carbon metabolism or the transsulfuration pathway. Folate cofactors carry and chemically activate single carbons for the synthesis of purines, thymidylate and methionine. So the evidence indicates that vitamin B6 plays an important role in maintenance of the genome, epigenetic stability and homocysteine metabolism. This article focuses on studies of strand breaks, micronuclei, or chromosomal aberrations regarding protective effects of vitamin B6, and probes whether it is folate-mediated one-carbon metabolism or the transsulfuration pathway for vitamin B6 which plays critical roles in prevention of cancer and cardiovascular disease.

Involvement of Pyridoxine/Pyridoxamine 5′- Phosphate Oxidase (PDX3) in Ethylene-Induced Auxin Biosynthesis in the Arabidopsis Root

  • Kim, Gyuree;Jang, Sejeong;Yoon, Eun Kyung;Lee, Shin Ae;Dhar, Souvik;Kim, Jinkwon;Lee, Myeong Min;Lim, Jun
    • Molecules and Cells
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    • v.41 no.12
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    • pp.1033-1044
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    • 2018
  • As sessile organisms, plants have evolved to adjust their growth and development to environmental changes. It has been well documented that the crosstalk between different plant hormones plays important roles in the coordination of growth and development of the plant. Here, we describe a novel recessive mutant, mildly insensitive to ethylene (mine), which displayed insensitivity to the ethylene precursor, ACC (1-aminocyclopropane-1-carboxylic acid), in the root under the dark-grown conditions. By contrast, mine roots exhibited a normal growth response to exogenous IAA (indole-3-acetic acid). Thus, it appears that the growth responses of mine to ACC and IAA resemble those of weak ethylene insensitive (wei) mutants. To understand the molecular events underlying the crosstalk between ethylene and auxin in the root, we identified the MINE locus and found that the MINE gene encodes the pyridoxine 5′-phosphate (PNP)/pyridoxamine 5′-phosphate (PMP) oxidase, PDX3. Our results revealed that MINE/PDX3 likely plays a role in the conversion of the auxin precursor tryptophan to indole-3-pyruvic acid in the auxin biosynthesis pathway, in which TAA1 (TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1) and its related genes (TRYPTOPHAN AMINOTRANSFERASE RELATED 1 and 2; TAR1 and TAR2) are involved. Considering that TAA1 and TARs belong to a subgroup of PLP (pyridoxal-5′-phosphate)-dependent enzymes, we propose that PLP produced by MINE/PDX3 acts as a cofactor in TAA1/TAR-dependent auxin biosynthesis induced by ethylene, which in turn influences the crosstalk between ethylene and auxin in the Arabidopsis root.

Isolation and Identification of Dextranase Production Strains and Enzyme Production (Dextranase 생산균주의 분리, 동정 및 효소생산)

  • Lee, Jong-Tae;Yi, Dong-Heui;Kwak, Yi-Seong;Kim, Young-Ho;Sung, Hyun-Soon;Kim, Chan-Jo
    • Microbiology and Biotechnology Letters
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    • v.23 no.4
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    • pp.405-410
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    • 1995
  • In order to screen dextranase with high dextranolytic activity from microbial origins, dextranase producing fungal isolates were isolated from soil of the Taeion area. 197 strains with dextranolytic activities were isolated, out of which 3 strains with high dextranolytic activities were selected in the first screening. A strain (GR-98) with a best dextranolytic activity was selected in the second screening. The strain was identified to be similiar Aspergillus ustus by the morphological and cultural characteristics. The optimum culture temperature and initial pH for the dextranase production of the strain was 30$\circ$C and 7.0, respectively. The optimum culture medium was composed of 2% dextran, 0.3% KNO$_{3}$, 0.05% K$_{2}$HPO$_{4}$, 0.02% MgSO$_{4}$-7H$_{2}$O, 0.05% KC1, and 2.5 $\mu$g/ml pyridoxamine, and the enzyme production was maximum when the strain was subcultured at 30$\circ$C for 7 days.

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Advanced Glycation Endproduct-induced Diabetic Complications

  • Lee, Hyun-Sun;Hong, Chung-Oui;Lee, Kwang-Won
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1131-1138
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    • 2008
  • Diabetic complications are a leading cause of blindness, renal failure, and nerve damage. Additionally, diabetes-accelerated atherosclerosis leads to increased risk of myocardial infarction, stroke, and limb amputation. At the present time, 4 main molecular mechanisms have been implicated in hyperglyceamia-mediated vascular damage. In particular, advanced glycation endproducts (AGE), which are formed by complex, heterogeneous, sugar-derived protein modifications, have been implicated as a major pathogenic process for diabetic complications. Recently, AGE inhibitors such as aminoguanidin, ALT-946, and pyridoxamine have been reported. Such an integrating paradigm provides a new conceptual framework for future research on diabetes complications and on discovering drugs to prevent the progression of AGE-induced maladies.

Catalytic and Structural Properties of Pyridoxal Kinase

  • Cho, Jung-Jong;Kim, Se-Kwon;Kim, Young-Tae
    • BMB Reports
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    • v.30 no.2
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    • pp.125-131
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    • 1997
  • This work reports studies of the catalytic and structural properties of pyridoxal kinase (ATP: pyridoxal 5' -phosphotransferase, EC. 2.7.1.35), Pyridoxal kinase catalyzes the phosphorylation of vitamin $B_6$ (pyridoxal, pyridoxamine, pyridoxine) using ATP-Zn as a phosphoryl donor. The enzyme purified from brain tissues is made up of two identical subunits of 40 kDa each. Native enzyme was inhibited by a substrate analogue, pyridoxal-oxime. Limited chymotrypsin digestion of pyridoxal kinase yields two fragments of 24 and 16 kDa with concomitant loss of catalytic activity. These fragments were isolated by DEAE ion exchange chromatography and used for binding studies with fluorescent ATP and pyridoxal analogues. The spectroscopic properties of both fluorescent pyridoxal analogue and Anthraniloyl ATP (Ant-ATP) bound to the 24 kDa fragment are indistinguishable from those of both pyridoxal analogue and Ant-ATP bound to the native pyridoxal kinase, respectively. The small 16 kDa fragment, generated by proteolytic cleavage of the kinase, does not bind any of the substrate analogues. Binding characteristics of Ant-ATP were extensively studied by measuring the changes in fluorescence spectra at various conditions. From the results presented herein, it is postulated that the structural domain associated with catalytic activity comprises approximately one-half of the molecular mass of pyridoxal kinase (24 kDa). whereas the remaining portion (16 kDa) of the enzyme contains a regulatory binding domain.

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$\alpha$-D-Glucosidase Inhibitor from Streptomyces sp. (II) -Cultural Conditions for the Inhibitor Production- (Streptomyces 속 균주가 생성하는 $\alpha$-D-Glucosidase Inhibitor(II)-저해물질의 생산조건 -)

  • 도재호;주현규
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.207-212
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    • 1989
  • Cultural conditions for $\alpha$-D-Glucosidase Inhibitor production from Streptomyces sp. YS-221-B isolated from soil arid identified as Streptomyces flauovirens or a subspecies of it were investigated. When the strain was cultured in a flask containing 2% glucose, 0.3% asparagine, 0.0002% riboflavin, 0.05% $K_2$HPO$_4$, 0.1% MgSO$_4$.7$H_2O$, 0.05% NaCl, pH 8.0 at 3$0^{\circ}C$, maximum production of the inhibitor was obtained after 8-9 days of cultivation. Sugar alcohols such as mannitol, i-inositol, erythritol as carbon sources, asparagine and beef extract as nitrogen sources were favorable for inhibitor production. Among vitamins, riboflavin, p-aminobenzoic acid, pyridoxamine and folic acid promoted the production of inhibitor, but depressed by the addition of hesperidine, and also depressed by cobalt, lithium and ferrous salts.

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Comparision of Preparation Methods for Water Soluble Vitamin Analysis in Foods by Reversed-Phase High Performance Liquid Chromatography (역상 고속 액체 크로마토그래피에 의한 식품 중 수용성 비타민 분석을 위한 전처리법의 비교)

  • Kim, Hyung-Soo;Jang, Duck-Kyu;Woo, Dong-Kyun;Woo, Kang-Lyung
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.141-150
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    • 2002
  • Owing to a need for simple extraction and purification for analysis of water soluble vitamins in food samples by RP-HPLC with UV-detector, the methods of bromelain and protease hydrolysis and $C_{18}$ Sep-Pak solid phase extraction were employed. The recoveries of standard water soluble vitamins by the bromelain and protease hydrolysis and $C_{18}$ Sep-Pak solid phase extraction were significantly high compared to AOAC methods in most of vitamins. The contents of pyridoxal determined with protest in the pork was similar, but in the bromelain hydrolysis and AOAC method, was high compared to the results of reference. The niacinamide, thiamin and riboflavin determined with bromelain and protease hydrolysis showed similar values to the results of references. In the potato, pyridoxamine was detected in the AOAC method, which was not detected in the bromelain and protease hydrolysis methods. Pyridoxal contents in the protease hydrolysis and AOAC methods were very similar to the results of references. The recoveries of fortified standard vitamins in food samples were significantly high and accurate compared to those of AOAC methods. The extraction and purification with $C_{18}$ Sep-Pak solid extractor might be considered superior method for the determination of water soluble vitamins in food samples.

Effects of Pasteurization and Frozen Storage on Changes in Quality Characteristics of 10% Salted Egg Yolk (저온살균 및 냉동저장이 10% 가염난황의 품질특성에 미치는 영향)

  • Kim, Jae-Wook;Choi, Chun-Un
    • Korean Journal of Food Science and Technology
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    • v.34 no.3
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    • pp.459-465
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    • 2002
  • To obtain the basic data for commercial 10% salted egg yolk for mayonnaise preparation, 3 types of egg yolks [pasteurized egg yolk (Yolk A)-not salted, pasteurized before salting (Yolk B)-salted, and pasteurized after salting (Yolk C)-salted] were prepared, and the changes in quality characteristics of these egg yolks with frozen storage were tested. The results obtained were as follows; Yolk A gelatinized during frozen storage, thus could not used for mayonnaise preparation. The viscosity of the egg yolk increased $3{\sim}5$ times after salting. Viscosity of the salted egg yolk increased with frozen storage time. Viscosity of Yolk B was higher at $-20^{\circ}C$ than $-15^{\circ}C$. Viscosity of Yolk C, however, was higher at $-15^{\circ}C$ than $-20^{\circ}C$. Frozen storage of pasteurized salted egg yolk showed some effects on the emulsification capacity. The effect, however, was smaller than that of unpasteurized salted egg yolk. Microbes of salted egg yolk were decreased with frozen storage, but there was no difference between Yolk B and Yolk C. It was suggested that commercially pasteurized 10% salted egg yolk for mayonnaise preparation can be successfully stored for 12 months at the temperature of $-15{\sim}-20^{\circ}C$.