• Journal of Nutrition and Health
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• 제19권4호
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• pp.246-254
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• 1986

이유한 Sprague Dawley 종의 암컷 흰쥐를 두군으로 나누어 각각 pyridoxine이 충분한 식이 (22mg/kg diet)와 pyridoxine이 부족한 식이 (1.2mg/kg diet0로 성장시킨후, 적절한 시기에 임신시켰다. 태어난 새끼가 이유할 때 결핍식이군을 둘로 나누어, 한쪽은 pyridoxine이 충분한 식이로 바꾸어준후, 세군의 쥐를 생후 10-주까지 성장시켰다. 각 군의 새끼쥐가 각각 0, 3, 7, 10 주 되었을대, 간의 mitochondria 및 cytosolic fraction 에 있는 Asparate aminotransferase [ EC 2.6.1.1] activity 및 pyridoxal phosphate의 함량을 측정하였다. 결핍군의 간 aspartate aminotransterase activity는 mitochondria 및 cytosolic fraction에서 각각 대조군에 비해 유의적으로 낮았으며 10-4 M pyridoxal phosphate를 첨가한 후 측정한 total enzyme activity 는 거의 대조군과 비슷하였다. 3 주이후 pyridoxine 이 충분한 식이로 바구어준 회복군에서도 대조군에 비해 유의적으로 낮았으나, 결핍군보다는 높았다. 이두 fraction 에서의 pyridoxal phosphate 함량은 결핍군이 대조군보다 현저히 낮았으며, 회복군은 대조군과 비슷하였다. 이로써, 장기간에 걸친 pyridoxine결핍이간의 aspartate aminotransferase activity 및 pyridoxal phosphate에 현저한 영향을 미치며 이유기 이후의 식이보충에 의해서도 enzyme activity는 쉽게 회복되지 않음을 알 수있다.

• BMB Reports
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• 제41권5호
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• pp.408-413
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• 2008

Pyridoxal-5'-phosphate phosphatase (PLPP) catalyzes the dephosphorylation of pyridoxal-5'-phosphate (PLP). A human brain PLPP gene was fused with a PEP-1 peptide and produced a genetic in-frame PEP-1-PLPP fusion protein. The purified PEP-1-PLPP fusion protein was efficiently transduced into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced PEP-1-PLPP fusion protein was stable for 36 h. The concentration of PLP was markedly decreased by the addition of exogenous PEP-1-PLPP to media pretreated with the vitamin $B_6$ precursors; pyridoxine, pyridoxal kinase and pyridoxine-5'-phosphate oxidase into cells. The results suggest that the transduction of the PEP-1-PLPP fusion protein can be one mode of PLP level regulation, and to replenish this enzyme in the various neurological disorders related to vitamin $B_6$.

• 한국식품과학회지
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• 제9권3호
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• pp.183-189
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• 1977

고정화(固定化) 균체(菌體)를 이용(利用)한 pyridoxal 5'-phosphate(pyridoxal-p)의 연속생산(連續生産)에 관(關)해실험(實驗)하였다. pyridoxine 5'-phosphate (pyridoxine-p) oxidase활성(活性)을 갖는 Pseudomonas polycolor 균체(菌體)와 Catalase 활성(活性)을 갖는 Kloeckera sp. No. 2201균체(菌體)를 효소원(酵素源)으로 사용(使用)하였다. 균체(菌體)는 Polyacrylamide gel에 불용화(不溶化) 시켰으며 동시고정균체(同時固定菌體)의 활성(活性)이 Pseudomonas Polycoler단독고정균체(單獨固定菌體)의 활성(活性)보다 강(强)하였다. 이 결과(結果)는 동시고정균체(同時固定菌體)의 Pyridoxine-p oxidase-cat-alase system은 Pyridoxine-p 산화(酸化)의 부산물(副産物)인 $H_2O_2$를 분해(分解)하므로서 보호효과를 얻을 수 있음을 의미(意味)한다. 동시고정균체(同時固定菌體)와 생균체(生菌體)의 효소활성(酵素活性)에 미치는 최적(最適)pH는 9.0이었고 동시고정균체(同時固定菌體)의 최적온도(最適溫度)는 $45^{\circ}C$로 생균체(生菌體)보다 $5^{\circ}C$높았다. 고정균체(固定菌體)의 Pyridoxine-p oxidase는 $Hg^{2+}$와 몇몇 SH-화합물(化合物)에 의(依)하여 활성화(活性化)되었다.

• BMB Reports
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• 제29권4호
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• pp.335-342
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• 1996

Porcine liver cysteinesulfinic acid decarboxylase was purified approximately 460-fold by means of ammonium sulfate fractionation and sequential column chromatographic separation with Sephadex G-100, DEAE-cellulose and hydroxylapatite. The enzyme has a flat pH profile with maximum activity occurring between pH 6.0 and 7.6. Pyridoxal 5'-phosphate must be present in all buffers used for purification procedures in order to stabilize the enzyme. Addition of sulfhydryl reagents such as 2-mercaptoethanol are also necessary to maintain maximum enzyme activity throughout purification. The absorption spectrum shows that cysteinesulfinic acid decarboxylase is a pyridoxal 5' -phosphate-containing protein. The major absorption is at 280 nm with two smaller absorption regions, one at 425 nm which is ascribed to a Schiffs base between pyridoxal phosphate and protein, and another at 325 nm which is thought to be due to the interaction of 2-mercaptoethanol with the Schiffs base. A number of divalent cations tested did not affect enzyme activity with the exception of mercury, copper, and zinc which are inhibitory. The partially purified enzyme has an apparent $K_m$ of 0.94 mM for cysteinesulfinate. Cysteic acid is a competitive inhibitor of the enzyme with a $K_i$ of 1.32 mM. The molecular weight of the enzyme was estimated to be about 79,600 by using Sephadex G-200 column chromatography.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1995년도 춘계학술대회
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• pp.74-74
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• 1995

GABA transaminase is inactivated by preincubation with p$^1$, p$^2$-bis(5'-pyridoxal) diphosphate at pH 7.0. The inactivation under pseudo-first order conditions proceeds at a slow rate (K$\_$obs/=0.035 min$\^$-1/). The degree of labeling of the enzyme by p$^1$, p$^2$-bis(5'-pyridoxal) diphosphate was determined by absorption spectroscopy, The blocking of 2 lysyl residues/dimer is needed for inactivation of the transaminase. The time course of the reaction is significantly affected by the substrate ${\alpha}$-ketoglutarate, which afforded complete protection against the loss of the catalytic activity. Whereas cofator pyridoxal phosphate failed to prevent the inactivation of the enzyme. Therefore, it is postulated that binding of ${\alpha}$-ketoglutarate tn lysyl residues is the major factor contributing to stabilization of the catalytic site and bifuctional reagent p$^1$, p$^2$bis(5'-pyridoxal) diphosphate blocks lysyl residues other than those involved in the binding of the cofactor.

• Journal of Nutrition and Health
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• 제28권4호
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• pp.321-330
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• 1995

Concentrations of total vitamin B-6 in human milk as well as individual, B-6 vitamers have important implications for the nutritional management of breast-fed(BF) infants. Vitamin B-6 status was assessed in 3 groups of infants : two groups preterm (PT) BF infants whose mothers were supplemented with 2 or 27mg pyridoxine(PN)-HCI ; a sub group of formula-fed (FF) PT infants. Mothers and infants were assessed weekly during the 28-day post feeding. Throughout the neonatal period, levels of total vitamin B-6 and percentages of pyridoxal(PL) in breast milk were lower in PT than T mothers, even in mothers supplemented with 27mg PN-HCI. Total vitamin B-6 levels in PT milk paralleled maternal supplementation but percentage distributions of B-6 vitamers did not change. Vitamin B-6 intakes of BF preterm infants paralleled their mothers' level of infants in the 2mg group was suggested by vitamin status parameters. Vitamin B-6 inadequacy of infants correlated with their plasma pyridoxal-5-phosphate(PLP) levels and erythrocyte alanine aminotransferase(E-ALAT) activity; all parameters such as plasma PLP, PL/PLP ratio and stimulation % of E-ALAT were highest for FF PT infants. The positive correlation of vitamin B-6 levels in breast milk gestational age may contraindicate its adequacy for some PT infants.

• Nutritional Sciences
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• 제3권1호
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• pp.31-35
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• 2000

The purpose of this study was to compare the changes in PLP concentrations induced by regular, moderate, and abrupt, high-intensity exercise in the plasma and tissues of vitamin B6 deficient and normal rats. Forty-eight rats were fed either a vitamin B6 deficient (-B6) diet or a normal (+B6) diet for 5 weeks and were subdivided into 4 groups:non-exercise(NE) group: regular, moderate-intensity exercise (RME) group; abrupt, high-intensity exercise (AIE) group; abrupt, high-intensity exercise and recuperation(IRE) group. The RME group was exercised on treadmill ($10^{\circ}$, 0.5-0.8km/h) for 2 hours just before sacrifice at the end of 5th week on the diet and the IER group was recuperated for three days on the diet after being exercised like the AIE group. Pyridoxal 5 -phosphate(PLP) levels were compared in the plasma, liver and skeletal muscle of the rats. Plasma PLP concentration tended to decrease in -B6 rats and tended to increase in +B6 rats with AIE. Plasma PLP concentration in both +B6 rats with AIE and no change in both -B6 and +B6 rats with RME. Muscle PLP concentration decreased in +B6 rats, showed no change in -B6 rats with AIE. Muscle PLP concentrations in both +B6 and -B6 rats did not change with RME. Plasma PLP, liver PLP and muscle PLP concentration of IER returned to those of NE in both +B6 and -B6 rats. These results suggest that changes in PLP concentration in plasma, liver and muscle occur with exercise and are affected by exercise intensity and vitamin B6 status. These changes may be due to interorgan redistribution of PLP.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.736-738
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• 2003

Ornithine decarboxylase (ODC) is the key enzyme in the synthetic pathway of polyamines. This enzyme is a homodimeric and a pyridoxal 5-phosphate (PLP) dependent enzyme. We have isolated, a cDNA clone encoding ODC from brain cDNA library constructed from flounder (Paralichthys olivaceus). The ODC cDNA contained a complete ORF consisting of 460 amino acids and one stop codon with comparison to nucleotide sequences of the flounder, zebrafish and rat ODC genes, the ODC genes were highly conserved. The transcription of ODC was analyzed with reverse transcription-polymerase chain reaction (RT-PCR) species in brain, kidney, liver, and embryo.

• BMB Reports
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• 제55권9호
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• pp.439-446
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• 2022

Pyridoxal 5'-phosphate (PLP)-dependent enzymes are ubiquitous, catalyzing various biochemical reactions of approximately 4% of all classified enzymatic activities. They transform amines and amino acids into important metabolites or signaling molecules and are important drug targets in many diseases. In the crystal structures of PLP-dependent enzymes, organic cofactor PLP showed diverse conformations depending on the catalytic step. The conformational change of PLP is essential in the catalytic mechanism. In the study, we review the sophisticated catalytic mechanism of PLP, especially in transaldimination reactions. Most drugs targeting PLP-dependent enzymes make a covalent bond to PLP with the transaldimination reaction. A detailed understanding of organic cofactor PLP will help develop a new drug against PLP-dependent enzymes.

• Journal of Nutrition and Health
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• 제28권5호
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• pp.426-434
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• 1995

The purpose of this study were to investigate the effect of fasting and vitamin B6 repletion on tissue concentration of pyridoxal 5-phosphate and urinary excreteion of 4-pyridoxic acid in vitamin B6 deficient rats. Sixty six rats(6 per group) were fed either a vitamin B6 deficient diet (-B6) or a control diet (+B6) for 6 weeks and then rats were repleted with +B6 diet for 2 weeks. Rats were fasted for 1 and 3 days and for 3 days after repletion. Pyridoxal 5-phosphate (PLP) concentration in plasma, liver, skeletal muscle, and heart muscle and urinary 4-pyridoxic acid (4-PA) excretion were compared. Fasting resulted in a significant increase in PLP concentration in the plasma, liver and heart muscle of rats fed the -B6 diet. Skeletal muscle PLP concentration was significantly decreased in +B6 rats but not in -B6 rats. Following vitamin B6 repletion, PLP concentration in the plasma, liver and heart muscle in previously -B6 rats was similar to the respective concentration in +B6 rats while PLP concentration in the skeletal muscle of previously -B6 rats increased, but it was not reached to that of +B6 rats. At day 1 and 2 of the fast, urinary 4-PT excretion increased in both +B6 and -B6 rats although there was no supply of vitamin B6 due to fasting. These results suggest that vitami B6 is redistributed as PLP when there is a caloric deficit and PLP is supplied by an endogenous source, possibly PLP bound to skeletal muscle glycogen phosphorylase.