• Preventive Nutrition and Food Science
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• v.14 no.4
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• pp.358-361
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• 2009

Aldose reductase was purified to electrophoretic homogeneity from porcine liver. The purified enzyme was a monomer of 36 kDa. The enzyme was strongly inhibited by $Cu^{2+}\;and\;Mg^{2+}$ ions. Incubation of the enzyme with pyridoxal 5'-phosphate led to complete inhibition of enzymatic activity, suggesting that lysine residue is involved at or near the active site of the enzyme. The enzyme exhibited a broad substrate specificity. Furthermore, the enzyme was capable of decolorizing Alizarin, an anthraquinone dye.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.270-270
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• 1994

돼지 뇌조직으로부터 순수 분리 정제된 Glutamate decarboxylase (GAD)는 효소 dimer당 0.8mole 보조 인자인 pyridoxal-5-phosphate(PLP)가 강하게 binding되어 있었다. 이러한 부분적으로 resolved된 효소에 외부로부터 PLP를 넣어주면 효소의 활성도는 최대값으로 증가하였다. 정제된 GAD는 sulfydryl시약에 의한 화학변형에 의하여 효소의 활성도를 상실하였으며 환원제인 dithiothreitol이나 2-mercaptoethanol의 첨가에 의하여 효소의 활성도가 복구되는 것으로 보아 효소의 활성부위의 활성에 직접 관여하는 중요한 cysteinyl잔기가 존재하고 있는 것을 알 수 있다.

• Journal of Microbiology and Biotechnology
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• v.33 no.5
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• pp.644-655
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• 2023

Safety assessment and functional analysis of probiotic candidates are important for their industrial applications. Lactiplantibacillus plantarum is one of the most widely recognized probiotic strains. In this study we aimed to determine the functional genes of L. plantarum LRCC5310, isolated from kimchi, using next-generation, whole-genome sequencing analysis. Genes were annotated using the Rapid Annotations using Subsystems Technology (RAST) server and the National Center for Biotechnology Information (NCBI) pipelines to establish the strain's probiotic potential. Phylogenetic analysis of L. plantarum LRCC5310 and related strains showed that LRCC5310 belonged to L. plantarum. However, comparative analysis revealed genetic differences between L. plantarum strains. Carbon metabolic pathway analysis based on the Kyoto Encyclopedia of Genes and Genomes database showed that L. plantarum LRCC5310 is a homofermentative bacterium. Furthermore, gene annotation results indicated that the L. plantarum LRCC5310 genome encodes an almost complete vitamin B₆ biosynthetic pathway. Among five L. plantarum strains, including L. plantarum ATCC 14917^T , L. plantarum LRCC5310 detected the highest concentration of pyridoxal 5'-phosphate with 88.08 ± 0.67 nM in MRS broth. These results indicated that L. plantarum LRCC5310 could be used as a functional probiotic for vitamin B₆ supplementation.

• Nutrition Research and Practice
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• v.8 no.6
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• pp.688-694
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• 2014

BACKGROUND/OBJECTIVES: Recent research regarding vitamin $B_6$ status including biochemical index is limited. Thus, this study estimated intakes and major food sources of vitamin $B_6$; determined plasma pyridoxal 5'-phosphate (PLP); and assessed vitamin $B_6$ status of Korean adults. MATERIALS/METHODS: Three consecutive 24-h diet recalls and fasting blood samples were collected from healthy 20- to 64-year-old adults (n = 254) living in the Seoul metropolitan area, cities of Kwangju and Gumi, Korea. Vitamin $B_6$ intake and plasma PLP were analyzed by gender and by vitamin $B_6$ supplementation. Pearson's correlation coefficient was used to determine associations of vitamin $B_6$ intake and plasma PLP. RESULTS: The mean dietary and total (dietary plus supplemental) vitamin $B_6$ intake was $1.94{\pm}0.64$ and $2.41{\pm}1.45mg/day$, respectively. Median (50th percentile) dietary intake of men and women was 2.062 and 1.706 mg/day. Foods from plant sources provided 70.61% of dietary vitamin $B_6$ intake. Only 6.3% of subjects consumed total vitamin $B_6$ less than Estimated Average Requirements. Plasma PLP concentration of all subjects was $40.03{\pm}23.71nmol/L$. The concentration of users of vitamin $B_6$ supplements was significantly higher than that of nonusers (P < 0.001). Approximately 16% of Korean adults had PLP levels < 20 nmol/L, indicating a biochemical deficiency of vitamin $B_6$, while 19.7% had marginal vitamin $B_6$ status. Plasma PLP concentration showed positive correlation with total vitamin $B_6$ intake (r = 0.40984, P < 0.0001). CONCLUSIONS: In this study, vitamin $B_6$ intake of Korean adults was generally adequate. However, one-third of subjects had vitamin $B_6$ deficiency or marginal status. Therefore, in some adults in Korea, consumption of vitamin $B_6$-rich food sources should be encouraged.

• Archives of Pharmacal Research
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• v.19 no.6
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• pp.480-485
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• 1996

Brain GABA transaminase is inactivated by preincubation with antidepressant/antipanic drug pheneizine (${\beta}$ethylphenylhydrazine) (mixing molar ratio 10:1) at pH 7.4. The reaction of enzyme with phenelzine was monitored by absorption and fluorescence spectroscopic methods. The inactive enzyme was fully reconstituted by addition of cofactor pyridoxal-5-phosphate. This result implies that the blocking of 1 mol of pyridoxal-5-phosphate per enzyme dimer is needed for inactivation of the enzyme. The time course of the reaction is significantly affected by the substrate .alpha.-ketoglutarate, which afforded complete protection against the loss of catalytic activity. The kinetic studies shows that phenelzine reacts with the cofactor of enzyme with a second-order rate constant of $2.1{\times}10^3M^{-1}s^{-1}$. It is postulated that the antidepressant/antipanic drug phenelzine is able to elevate the neurotransmitter GABA levels in central nervous system by inhibitory action on GABA degradative enzyme GABA transaminase.

• Applied Biological Chemistry
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• v.22 no.4
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• pp.191-197
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• 1979

${\beta}-Tyrosinase$ was purified and crystallized from cells of Escherichia intermedia A-21 grown in a medium supplemented with 0.2% L-tyrosine. Molecular weight of its subunit, Km value and absorption spectra were determined. Crystallization methods were also studied to eliminate any unnecessary procedures. The results obtained were as follows: 1. The purification procedure included ammonium sulfate fractionation, dialysis against potassium phosphate buffer, pH 6.0 and pH 7.0, and DEAE-Sephadex column chromatography. In the column chromatography, 11 mg of protein was applied per ml of DEAE-Sephadex for efficiency. 2. Steps of protamine sulfate treatment and Sephadex G-150 gel filtration could be eliminated for this enzyme from the known procedures. 3. The purified enzyme was dissolved in 0.01M potassium phosphate buffer containing 2-mercaptoethanol, with a concentration of 20mg/ml. Crystalline enzyme, which appears as hexagonal rods, was obtained by adding solid fine powdered ammonium sulfate to the enzyme solution. 4. Absorption maxima of the enzyme appeared at 340 and 430nm when associated with pyridoxal phosphate. 5. Km value of the enzyme for L-tyrosine was $2.31{\times}10^{-4}M$ and the molecular weight of its subunit was determined by SDS-polyacrylamide electrophoresis to be approximately 50,000.

• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.4
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• pp.216-220
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• 2002

1,4-Benzoquinone reductase was purified to electrophoretic homogeneity from bovine liver, and the purified enzyme found to have a molecular mass of 29 kDa, as determined by sodium dodecyl sulfate- polyacrylamide gel electrophoresis The enzyme exhibited pH optimum between 8.0 and 8.5. The apparent fm for 1,4-benzoqulnone was 1.643 mM, and the apparent Km for NADH was 1.837 mM. Various divalent cations, such as Hg$\^$2+/, Cu$\^$2+/, and Zn$\^$2+/, exhibited strong inhibitory effects. The enzyme activity was also strongly inhibited by quercetin, dicumarol, and benzoic acid. Incubation of the enzyme with N-bromosuccinimide and pyridoxal 5’-phosphate led to inhibitions of 100% and 99%, respectively. Accordingly, these results suggest that trypto-phan and Iysine residues are Involved at or near the active sites of the enzyme.

• Journal of Nutrition and Health
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• v.26 no.7
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• pp.867-886
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• 1993

Vitamin B-6 status parameters of mothers were assessed in relation to th condition of the infant at birth and during the neonatal period. Parameters were assessed at birth and then weekly in 18 mother-infant pairs during the neonatal period ; mothers were supplemented postnatally with 2 or 27 mg PN-HCI/d. Vitamin B-6 inadequacy in the 2mg supplemented group was suggested by the vitamin status parameters. Mothers whose infants had unsatisfactory Apgar scores at 5min after birth(<7) had lower vitamin B-6 status parameters than mothers whose infants were scored satisfactory. Also, infants who scored unsatisfactory at birth and whose mothers were supplemented with the low level of PN had significantly lower vitamin B-6 status parameters at 7 days of age than infants who scored satisfactory. Infants scored unsatisfactory showed some beneficial effects in both vitamin B-6 status and growth associated with the higher level of maternal postnatal vitamin B-6 supplement. In summary, the mother's prenatal and postnatal vitamin B-6 intake were significantly related to the condition of her infant at birth and during the neonatal period, respectively.

• Journal of Microbiology
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• v.35 no.2
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• pp.97-102
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• 1997

Cysteine desulfhydrase (EC 4.4.1.1.) was purified from the culture supernatant of Streptomyces albidoflavus SMF301 by hydroxyapatite, gel filtration and Resource Q ion-exchange chromatography with a purification fold of six identical subunits. The enzyme was stabilized by dithiothreitol and pyridoxal 5'-phosphate during the purification procedures. The optimum pH and temperature were pH 8.6 and 35$^{\circ}C$, respectively. The N-terminal amino acid sequence was identified as A-P-L-P-T-A-D-V-R-S-D-P-G-Y-R-E-W-L-G-E-A-V. The purified cystein desulfhydrase had a high substrate specificity toward cysteine, and exhibited no cystahionine $\gamma$-lyase activity. The $K_m$ value for cysteine was determined to be 0.37 mM.