• 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.

• The Plant Pathology Journal
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• 제37권6호
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• pp.673-680
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• 2021

Acidovorax citrulli (Ac) is the causative agent of bacterial fruit blotch disease in watermelon. Since resistant cultivars have not yet been developed, the virulence factors/mechanisms of Ac need to be characterized. This study reports the functions of a putative pyridoxal phosphate-dependent aminotransferase (PpdaAc) that transfers amino groups to its substrates and uses pyridoxal phosphate as a coenzyme. It was observed that a ppdaAc knockout mutant had a significantly reduced virulence in watermelon when introduced via germinated-seed inoculation as well as leaf infiltration. Comparative proteomic analysis predicted the cellular mechanisms related to PpdaAc. Apart from causing virulence, the PpdaAc may have significant roles in energy production, cell membrane, motility, chemotaxis, post-translational modifications, and iron-related mechanisms. Therefore, it is postulated that PpdaAc may possess pleiotropic effects. These results provide new insights into the functions of a previously unidentified PpdaAc in Ac.

• 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.

• The Plant Pathology Journal
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• 제39권3호
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• pp.235-244
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• 2023

Acidovorax citrulli (Ac) is a phytopathogenic bacterium that causes bacterial fruit blotch (BFB) in cucurbit crops, including watermelon. However, there are no effective methods to control this disease. YggS family pyridoxal phosphate-dependent enzyme acts as a coenzyme in all transamination reactions, but its function in Ac is poorly understood. Therefore, this study uses proteomic and phenotypic analyses to characterize the functions. The Ac strain lacking the YggS family pyridoxal phosphate-dependent enzyme, AcΔyppAc(EV), virulence was wholly eradicated in geminated seed inoculation and leaf infiltration. AcΔyppAc(EV) propagation was inhibited when exposed to L-homoserine but not pyridoxine. Wild-type and mutant growth were comparable in the liquid media but not in the solid media in the minimal condition. The comparative proteomic analysis revealed that YppAc is primarily involved in cell motility and wall/membrane/envelop biogenesis. In addition, AcΔyppAc(EV) reduced biofilm formation and twitching halo production, indicating that YppAc is involved in various cellular mechanisms and possesses pleiotropic effects. Therefore, this identified protein is a potential target for developing an efficient anti-virulence reagent to control BFB.

• 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.

• 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.

• 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.