• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.10a
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• pp.14-14
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• 2010

Vitamin C (ascorbic acid) is an essential component for collagen biosynthesis and also for the proper functioning of the cardiovascular system in humans. Unlike most of the animals, humans lack the ability to synthesize ascorbic acid on their own due to a mutation in the gene encoding the last enzyme of ascorbate biosynthesis. As a result, vitamin C must be obtained from dietary sources like plants. In this study, we have developed two different kinds of transgenic potato plants (Solanumtuberosum L. cv. Taedong Valley) overexpressing strawberry GalUR and mouse GLoase gene under the control of CaMV 35S promoter with increased ascorbic acid levels. Integration of the these genes in the plant genome was confirmed by PCR and Southern blotting. Ascorbic acid(AsA) levels in transgenic tubers were determined by high-performance liquid chromatography(HPLC). The over-expression of these genes resulted in 2-4 folds increase in AsA intransgenic potato and the levels of AsA were positively correlated with increased geneactivity. The transgenic lines with enhanced vitamin C content showed enhanced tolerance to abiotic stresses induced by methyl viologen(MV), NaCl or mannitol as compared to untransformed control plants. The leaf disc senescence assay showed better tolerance in transgenic lines by retaining higher chlorophyll as compared to the untransformed control plants. Present study demonstrated that the over-expression of these gene enhanced the level of AsA in potato tubers and these transgenics performed better under different abiotic stresses as compared to untransformed control. We have also investigated the mechanism of the abiotic stress tolerance upon enhancing the level of the ascorbate in transgenic potato. The transgenic potato plants overexpressing GalUR gene with enhanced accumulation of ascorbate were investigated to analyze the antioxidants activity of enzymes involved in the ascorbate-glutathione cycle and their tolerance mechanism against different abiotic stresses under invitro conditions. Transformed potato tubers subjected to various abiotic stresses induced by methyl viologen, sodium chloride and zinc chloride showed significant increase in the activities of superoxide dismutase(SOD, EC 1.15.1.1), catalase, enzymes of ascorbate-glutathione cycle enzymes such as ascorbate peroxidase(APX, EC 1.11.1.11), dehydroascorbate reductase(DHAR, EC 1.8.5.1), and glutathione reductase(GR, EC 1.8.1.7) as well as the levels of ascorbate, GSH and proline when compared to the untransformed tubers. The increased enzyme activities correlated with their mRNA transcript accumulation in the stressed transgenic tubers. Pronounced differences in redox status were also observed in stressed transgenic potato tubers that showed more tolerance to abiotic stresses when compared to untransformed tubers. From the present study, it is evident that improved to lerance against abiotic stresses in transgenic tubers is due to the increased activity of enzymes involved in the antioxidant system together with enhanced ascorbate accumulated in transformed tubers when compared to untransformed tubers. At moment we also investigating the role of enhanced reduced glutathione level for the maintenance of the methylglyoxal level as it is evident that methylglyoxal is a potent cytotoxic compound produced under the abiotic stress and the maintenance of the methylglyoxal level is important to survive the plant under stress conditions.

• Food Science of Animal Resources
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• v.41 no.5
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• pp.840-854
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• 2021

The impact of green tea extract powder and rosemary extract powder, alone or in combination, on the quality characteristics of naturally cured pork sausages produced with white kimchi powder as a nitrate source was evaluated. Ground pork sausages were assigned to one of seven treatments: control (0.01% sodium nitrite and 0.05% sodium ascorbate), treatment 1 (0.3% white kimchi powder and 0.05% green tea extract powder), treatment 2 (0.3% white kimchi powder and 0.1% green tea extract powder), treatment 3 (0.3% white kimchi powder and 0.05% rosemary extract powder), treatment 4 (0.3% white kimchi powder and 0.1% rosemary extract powder), treatment 5 (0.3% white kimchi powder, 0.05% green tea extract powder, and 0.05% rosemary extract powder), and treatment 6 (0.3% celery juice powder, 0.05% green tea extract powder, and 0.05% rosemary extract powder). Naturally cured products had lower (p<0.05) cooking yield and residual nitrite content than control sausages. However, compared to the control, naturally cured products with white kimchi powder (treatments 1 to 5) showed similar the pH, oxidation-reduction potential, CIE L^* values, CIE a^* values, nitrosyl hemochrome content, total pigment content, and curing efficiency to the control. When the amount of green tea extract powder or rosemary extract powder was increased to 0.1% (treatments 2 and 4), lipid oxidation was reduced (p<0.05). These results indicate that green tea extract powder, rosemary extract powder, and white kimchi powder may provide an effective solution to replace synthetic nitrite and ascorbate used in traditionally cured products.

• Korean Journal of Food Science and Technology
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• v.19 no.6
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• pp.506-514
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• 1987

Peroxidase was purified to a homogeneous state from Castanea Semen by ammonium sulfate precipitation, DEAE-cellulose column chromatography, gel filtration on sephadex G-100 and HPLC, and the purification fold was 65.3. The molecular weight of the enzyme was estimated to be about 35,000 by HPLC. In properties of the enzyme which was purified up to sephadex G-100 column chromatography, the optimum pH and temperature were 5.0 and $50^{\circ}C$, respectively. By heating the enzyme at $80^{\circ}C$ for 1.73 min., the enzyme activity was decreased to 10%. The enzyme was active toward aromatic amines such as o-phenylenediamine and p-phenylendiamine. Kinetic studies indicated a Km of 2.6mM for o-phenylenediamine at an optimal hydrogen-peroxide concentration and a Km of 10mM for hydrogenperoxide at an optimal o-phenylenediamine concentration. Among the reagents tested, L-ascorbic acid and sodium L-ascorbate inhibited significantly the enzyme, while $Ca^{++}$ and $Ba^{++}$ activated the enzyme at the concentration of 1mM and 5mM.

• Journal of the Korean Home Economics Association
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• v.19 no.2
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• pp.183-188
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• 1981

This study was conducted to investigate the effects of synthetic antioxidants con the degradation of angiotensin II which is made up of 8 amino acids: Asp-Arg-Val-Try-Ile-Gly-Pro-Phe, by the $\alpha$-chymotrypsin and trypsin. the results obtained were as follow; 1. Dibutyl hydroxytoluene, butyl hydroxyanisole and sodium L-ascorbate showed no inhibitory effect on the activity of $\alpha$-chymotrypsin on the angiotensin II, but ethyl protocathechuate inhibited. its activity at the concentration of 100ppm. However, the angiotension II was gradually degradated by $\alpha$-chymotrypsin after one hour incubation with ethylprotecathechuate. 2. Butyl hydroxyanisole inhibited trypsin activities above 100ppm, but no inhibitory activities was observed by the other antioxidants used in this experiment.

• Journal of Pharmaceutical Investigation
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• v.29 no.3
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• pp.205-210
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• 1999

To formulate the parenteral delivery of a new cephalosporin derivative, 7-${\beta}$-[(2)-2-(2-arninothiazol-4-yl)-2methoxyiminoacetamido]- 3- [(2,3-cyclopenteno-4-carbamoyl-l-pyridinium)methyl]- 3-cephem-4-carboxylate sulfate( CKD604), the stability and solubility of CKD-604 in various aqueous media were investigated. The degradation kinetics of CKD-604 in aqueous solutions (ionic strength 0.1, pH 1-8) were studied at $37^{\circ}C$. The observed degradation rates followed pseudo first order kinetics. The pH-rate profile exhibited a minimum degradation rate at pH 5. The Arrhenius activation energy was 14.2 kcal/mol in pH 5 buffer solution. Excellent agreement between the cephalosporins' theoretical pH-rate profile and the experimental data indicated that the degradation pathway of CKD-604 could be predicted according to the general pathway of cephalosporins. The solubility of CKD-604 was 8.16 mg/ml at $25^{\circ}C$. To enhance the solubility and adjust the suitable pH, CKD-604 was solubilized by using sodium ascorbate, ascorbic acid and urea. The compositions were obtained to satisfy optimum pH and concentration, and the total amount of additives was several times of the active ingredient, CKD-604.

• Plant Biotechnology Reports
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• v.2 no.2
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• pp.113-122
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• 2008

Nitric oxide (NO) affects the growth and development of plants and also affects plant responses to various stresses. Because NO induces root differentiation, we examined whether or not it is involved in increased ROS generation. Treatments with sodium nitroprusside (SNP), an NO donor, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO), a specific NO scavenger, and $N{\omega}-nitro-{\text\tiny{L}}-arginine$ methyl ester hydrochloride (${\text\tiny{L}}-NAME$), an NO synthase (NOS) inhibitor, revealed that NO is involved in the adventitious root growth of mountain ginseng. Supply of an NO donor, SNP, activates NADPH oxidase activity, resulting in increased generation of $O_2{^{{\cdot}-}}$, which subsequently induces growth of adventitious roots. Moreover, treatment with diphenyliodonium chloride (DPI), an NADPH oxidase inhibitor, individually or with SNP, inhibited root growth, NADPH oxidase activity, and $O_2{^{{\cdot}-}}$ anion generation. Supply of the NO donor, SNP, did not induce any notable isoforms of enzymes; it did, however, increase the activity of pre-existing bands of NADPH oxidase, superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, and glutathione reductase. Enhanced activity of antioxidant enzymes induced by SNP supply seems to be responsible for a low level of $H_2O_2$ in the adventitious roots of mountain ginseng. It was therefore concluded that NO-induced generation of $O_2{^{{\cdot}-}}$ by NADPH oxidase seems to have a role in adventitious root growth of mountain ginseng. The possible mechanism of NO involvement in $O_2{^{{\cdot}-}}$ generation through NADPH oxidase and subsequent root growth is discussed.

• Journal of Life Science
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• v.25 no.5
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• pp.487-495
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• 2015

We carried out pH stability, chemical inhibition, chemical modification, and pH-dependent kinetic parameter assessments to further characterize protocatechuate 3,4-dioxygenase from Pseudomonas pseudoalcaligenes KF707. Protocatechuate 3,4-dioxygenase was stable in the pH range of 4.5~10.5. L-ascorbate and glutathione were competitive inhibitors with $K_{is}$ values of 0.17 mM and 0.86 mM, respectively. DL-dithiothreitol was a noncompetitive inhibitor with a $K_{is}$ value of 1.57 mM and a $K_{ii}$ value of 8.08 mM. Potassium cyanide, p-hydroxybenzoate, and sodium azide showed a noncompetitive inhibition pattern with $K_{is}$ values of 55.7 mM, 0.22 mM, and 15.64 mM, and $K_{ii}$ values of 94.1 mM, 8.08 mM, and 662.64 mM, respectively. $FeCl_{2}$ was the best competitive inhibitor with a $K_{is}$ value of $29{\mu}M$. $FeCl_{3}$, $MnCl_{2}$, $CoCl_{2}$, and $AlCl_{3}$ were also competitive inhibitors with $K_{is}$ values of 1.21 mM, 0.85 mM, 3.98 mM, and 0.21 mM, respectively. Other metal ions showed noncompetitive inhibition patterns. The pH-dependent kinetic parameter data showed that there may be at least two catalytic groups with pK values of 6.2 and 9.4 and two binding groups with pK values of 5.5 and 9.0. Lysine, cysteine, tyrosine, carboxyl, and histidine were modified by their own specific chemical modifiers, indicating that they are involved in substrate binding and catalysis.

• Journal of Environmental Science International
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• v.19 no.12
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• pp.1323-1334
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• 2010

The effect of nitric oxide (NO) on antioxidant system and protective mechanism against oxidative stress under UV-B radiation was investigated in leaves of maize (Zea mays L.) seedlings during 3 days growth period. UV-B irradiation caused a decrease of leaf biomass including leaf length, width and weight during growth. Application of NO donor, sodium nitroprusside (SNP), significantly alleviated UV-B stress induced growth suppression. NO donor permitted the survival of more green leaf tissue preventing chlorophyll content reduction and of higher quantum yield for photosystem II than in non-treated controls under UV-B stress, suggesting that NO has protective effect on chloroplast membrane in maize leaves. Flavonoids and anthocyanin, UV-B absorbing compounds, were significantly accumulated in the maize leaves upon UV-B exposure. Moreover, the increase of these compounds was intensified in the NO treated seedlings. UV-B treatment resulted in lipid peroxidation and induced accumulation of hydrogen peroxide ($H_2O_2$) in maize leaves, while NO donor prevented UV-B induced increase in the contents of malondialdehyde (MDA) and $H_2O_2$. These results demonstrate that NO serves as antioxidant agent able to scavenge $H_2O_2$ to protect plant cells from oxidative damage. The activities of two antioxidant enzymes that scavenge reactive oxygen species, catalase (CAT) and ascorbate peroxidase (APX) in maize leaves in the presence of NO donor under UV-B stress were higher than those under UV-B stress alone. Application of 2-(4-carboxyphenyl)-4, 4, 5, 5-tetramethylimidazoline-1-oxyl-3- oxide (PTIO), a specific NO scavenger, to the maize leaves arrested NO donor mediated protective effect on leaf growth, photosynthetic pigment and free radical scavenging activity. However, PTIO had little effect on maize leaves under UV-B stress compared with that of UV-B stress alone. $N^{\omega}$-nitro-L-arginine (LNNA), an inhibitor of nitric oxide synthase (NOS), significantly increased $H_2O_2$ and MDA accumulation and decreased antioxidant enzyme activities in maize leaves under UV-B stress. This demonstrates that NOS inhibitor LNNA has opposite effects on oxidative resistance. From these results it is suggested that NO might act as a signal in activating active oxygen scavenging system that protects plants from oxidative stress induced by UV-B radiation and thus confer UV-B tolerance.

• Food Science of Animal Resources
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• v.31 no.4
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• pp.588-595
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• 2011

In this study, the effects of red ginseng extracts (RGE), which has been used as an antioxidant and antimicrobial agent, on pork sausage were evaluated. The treatments were as follows; addition of 0.01% sodium ascorbate (V), 0.5% RGE (T1), 1.0% RGE (T2) and 1.5% RGE (T3) to the basal formula (C). T3 had a significantly higher pH, cooking loss and yellowness ($CIEb^*$) and lower lightness (CIE $L^*$) and redness (CIE $a^*$) than the other samples. The hardness and surface hardness values of 1.5% RGE treated sample were significantly lower (p<0.01) than those of C. However, the cohesiveness values of the RGE samples were higher than the others (p<0.05). In the sensory evaluation, no significant differences in color, taste, texture, juiciness and acceptability were observed among the tested samples, while, the aroma scores of T2 and T3 were higher than those of the C and V samples (p<0.05). The TBARS values of RGE treated groups were higher (p<0.05) than the C sample after 1, 2 and 3 weeks of storage; however, this value did not increase with storage time (p>0.05). When the RGE concentration was high, the reduction in total plate counts and VBN value at week 3 and 4 of storage (p<0.01) decreased. In conclusion, red ginseng extracts seemed to have a positive impact on lipid oxidation, aroma and the microbial characteristics of pork sausage.

• Food Science of Animal Resources
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• v.31 no.5
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• pp.710-718
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• 2011

The effects of organic acids mix (0.4%) and modified atmosphere packaging (MAP) on the storage quality of sliced bacon were investigated. Pork bellies were treated with or without organic acids at the curing stage. The organic acids mix comprised 35% sodium acetate, 25% salt, 15% calcium lactate, 11% trisodium citrate, 7% ascorbate, and 7% citric acid. The cured pork bellies were smoked and packaged with 50% $CO_2$ + 50% $N_2$ (50% $CO_2$-MAP) and 100% $N_2$ (100% $N_2$-MAP), and stored at $5^{\circ}C$ for 14 d. The 50% $CO_2$-MAP showed a higher pH value (p<0.05) up to 10 d, a lower protein deterioration (p<0.05) as measured by volatile basic nitrogen (VBN) from 6 to 14 d, and a higher color value of lightness (CIE $L^*$) compared to 100% $N_2$-MAP. The development of lipid oxidation measured by thiobarbituric acid reactive substance (TBARS) values seemed to be effectively controlled throughout the storage period in both 50% $CO_2$-MAP and 100% $N_2$-MAP regardless of the application of organic acids. The 50% $CO_2$-MAP inhibited the growth of aerobic and anaerobic bacteria (p<0.05) both in non-added and bacon added with organic acids mix. The 50% $CO_2$-MAP alone seemed to be effective in delaying the growth of bacteria since the use of organic acids mix gave no additional effects. The addition of organic acids mix lowered the pH value (p<0.05), effectively retarded the protein deterioration (p<0.05), and showed a higher color value of lightness (CIE $L^*$) value (p<0.05) and lower color value of redness (CIE $a^*$) value (p<0.05). In conclusion, 50% $CO^2$-MAP showed better quality and self-life of sliced bacon during storage. However, the beneficial effect of organic acids mix was not noticed in the concentration used in this experiment.