• KSBB Journal
• /
• v.5 no.2
• /
• pp.191-194
• /
• 1990

Tyrosine is a monohydrolic aromatic amino acid and DOPA is a tyrosine derivative containing dihydroxy group. DOPA can be synthesized from tyrosine by enzymatic reaction. The separation and quantitative determination of each component are very difficult in the reaction mixture. In the present study, two wavelengths giving maximum absorbance difference of each amino acid were determined using UV/VIS spectrophotometer by wavelength scanning and simple assay method was developed for the analysis of the reaction mixture of tyrosine and DOPA by measuring absorbances of reaction mixture. This method can be simply used for the analysis of the tyrosine and DOPA mixture because it does not require and procedure for the pretreatment of the reaction mixture.

• Nutrition Research and Practice
• /
• v.7 no.4
• /
• pp.249-255
• /
• 2013

In this study, the protective effects of EGCG on L-3,4-dihydroxyphenylalanine (L-DOPA)-induced oxidative cell death in catecholaminergic PC12 cells, the in vitro model of Parkinson's disease, were investigated. Treatment with L-DOPA at concentrations higher than $150{\mu}M$ caused cytotoxicity in PC12 cells, as determined using the 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry detection. The apoptotic ratio was similar in cells treated with $100{\mu}M$ EGCG plus $150{\mu}M$ L-DOPA (5.02%) and the control (0.96%) (P > 0.05), and was lower than that of cells treated with L-DOPA only (32.24%, P < 0.05). The generation level of ROS (% of control) in cells treated with EGCG plus L-DOPA was lower than that in cells treated with L-DOPA only (123.90% vs 272.32%, P < 0.05). The optical density in production of TBARS in cells treated with L-DOPA only was higher than that in the control ($0.27{\pm}0.05$ vs $0.08{\pm}0.04$, P < 0.05), and in cells treated with EGCG only ($0.14{\pm}0.02$, P < 0.05), and EGCG plus L-DOPA ($0.13{\pm}0.02$, P < 0.05). The intracellular level of GSH in cells treated with EGCG plus L-DOPA was higher than that in cells treated with L-DOPA only ($233.25{\pm}16.44$ vs $119.23{\pm}10.25$, P < 0.05). These results suggest that EGCG protects against L-DOPA-induced oxidative apoptosis in PC12 cells, and might be a potent neuroprotective agent.

• Journal of Life Science
• /
• v.28 no.7
• /
• pp.802-810
• /
• 2018

Aging is accompanied by changes in the body, such as graying hair, wrinkles, and black spots composed of lipid peroxides and proteins. Melanin is a polymer substance produced by an oxidation polymerization reaction from tyrosine, and it determines the color of hair and skin. It has been reported that melanin is synthesized by melanocyte, and its excessive production by reactive oxygen species is associated with aging. The purpose of this study was to determine the direct effects of Musa paradisiaca peel ethanolic extract (MPEE) on antioxidative activity and melanin synthesis. It was observed that the antioxidant activity of MPEE was similar to that of vitamin C, a positive control, in both DPPH radical scavenging assay and reducing power assay. In order to examine cytotoxicity prior to cell experimentation, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed for B16F1 cells. MPEE was not cytotoxic at $32{\mu}g/ml$ or less. In addition, MPEE increased melanin synthesis in live cells in addition to tyrosinase activity and melanin synthesis in dihydroxyphenylalanine (DOPA)-oxidation assay in vitro. Moreover, MPEE increased melanin synthesis in cells aged by pretreatment with $H_2O_2$. The expression levels of tyrosinase-related protein (TRP)-1, TRP-2, and superoxide dismutase (SOD)-2 by western blot analysis were increased in the presence of MPEE. These results suggest that MPEE could promote the melanin synthesis as an antioxidative substance.

• Biomolecules & Therapeutics
• /
• v.23 no.3
• /
• pp.283-289
• /
• 2015

The half-dried leaves of Stewartia. pseudocamellia were extracted with hot water (SPE) and partitioned with n-hexane (SPEH), dichloromethane (SPED), and ethyl acetate (SPEE) successively. SPE and SPEE showed significant inhibitory effects against melanogenesis and tyrosinase activities. By bioassay-guided isolation, ten phenolic compounds were isolated by column chromatography from SPEE. The whitening effect of the isolated compounds from SPEE were tested for the inhibitory activities against melanogenesis using B16 melanoma cells, in vitro inhibition of tyrosinase, and L-3,4-dihydorxy-indole-2-carboxylic acid (L-DOPA) auto-oxidation assay. A cytotoxic activity assay was done to examine the cellular toxicity in Raw 264.7 macrophage cells. Of the compounds isolated, gallic acid and quercetin revealed significant inhibitory activities against melanogenesis compared to arbutin. In particular, quercetin exhibited similar inhibitory activities against tyrosinase and L-DOPA oxidation without cytotoxicity. These results suggested that SPE could be used as a potential source of natural skin-whitening material in cosmetics as well as in food products.

• The Korean Journal of Mycology
• /
• v.38 no.2
• /
• pp.167-171
• /
• 2010

The present study aims to evaluate Cordyceps militaris water extract (CMWE) with a view to develop melanogenesis inhibitors. Inhibitory activities of CMWE against tyrosinase, L-DOPA(L-3,4-dihydroxyphenylalanine) oxidation, and melanin biosynthesis in B16 mouse melanoma cells were investigated. CMWE, at $5000\;{\mu}g/ml$, inhibited tyrosinase activity of 71% and DOPA oxidation of 40% as reacting with L-DOPA. Furthermore, B16 mouse melanoma cell survived over 50% from low to high dose on MTT assay, and CMWE markedly inhibited (> 50%) melanin synthesis at $5000\;{\mu}g/ml$. The inhibitory effect of CMWE on melanogenesis was attributed to enhancement of tyrosinase degradation. Key enzyme of melanin biosynthesis is tyrosinase which catalyses a beginning step from tyrosine to DOPA quinine and melanin formation step, respectively. These results indicated that CMWE may be a potential source of novel whitening agents for cosmetic or therapeutic application.

• Journal of Life Science
• /
• v.28 no.3
• /
• pp.331-338
• /
• 2018

Melanin is a polymer substance that plays an important role in the determination of hair growth and skin color in vivo. However, melanin, which is over-produced by reactive oxygen species, is known to cause stains, freckles, and hypercholesterolemia, which are associated with aging. Previous studies have shown that polyphosphate, one of the components of Rhynchosia Nulubilis, inhibits skin aging induced by ultraviolet rays. The aim of this study is to investigate the direct effect of Rhynchosia Nulubilis ethanolic extract (RNEE) on melanin synthesis. In this study, RNEE showed no antioxidative effects on scavenging activity of DPPH radical in addition to reducing power. The cytotoxicity of RNEE was increased in a dose-dependent manner in an MTT assay. In addition, RNEE increased tyrosinase activity and melanin synthesis in DOPA-oxidation experiments. RNEE did not promote the conversion L-DOPA into melanin in live cells, but melanin production was promoted in the RNEE-treated group after H2O2 pretreatment compared to the control group in which melanin production was reduced by treatment with H2O2. In addition, RNEE increased the expression level of tyrosinase related protein-2 (TRP-2) and increased the expression level of tyrosinase related protein-1 (TRP-1) at a concentration of $16{\mu}g/ml$. In particular, it was found that RNEE increased the expression level of SOD-3, by which superoxide anion is converted to hydrogen peroxide, higher than the control and ${\alpha}$-MSH used as a positive control at a concentration of more than $16{\mu}g/ml$. The results suggest that RNEE can induce melanogenesis related to black hair.

• Journal of Plant Biotechnology
• /
• v.46 no.1
• /
• pp.45-55
• /
• 2019

TheCitrus grandis Osbeck is a special product in the Jeju island. The product has been as a remedy for liver damage and hang over. This study demonstrates how to investigate and compare the antioxidant, phenol content, tyrosinase and ${\alpha}$-glucosidase inhibitory activity, antimicrobial, and alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activity with the C. grandis leaves extracted in different ethanol concentrations. From the yield, a 20% ethanol extract demonstrated the highest results among the other extracts. The distilled water extract showed the most abundant in a total phenol content and highest ABTS radical scavenging activity and reducing power assay. In the DPPH radical scavenging activity, ${\alpha}$-glucosidase and tyrosinase inhibitory assay (used ${\text\tiny{L}}$-tyrosine as substrate), the 80% ethanol extract exhibited a higher value than other extracts. The 60% ethanol extract showed prominent activities in the tyrosinase inhibitory (used ${\text\tiny{L}}$-dopa as substrate), ADH and ALDH activity assay. In the minimum inhibitory concentration (MIC) assay, 60% and 80% ethanol extracts inhibited the bacterial growth almost similarly. Moreover, the gram-positive bacteria was more restrained than the gram-negative bacteria. The resultsrevealed that the distilled water and 80% ethanol extract showed a relatively higher antioxidant activity compared to other extracts. The 60 ~ 80% ethanol extracts demonstrated potential tyrosinase, ${\alpha}$-glucosidase inhibitory, antimicrobial, ADH and ALDH activities. Therefore, the C. grandis is suggested to be considered as a functional material for various proposes.

• Horticultural Science & Technology
• /
• v.29 no.4
• /
• pp.358-365
• /
• 2011

Lespedeza cuneata G. Don is a plant commonly grown in Asian countries, which has been widely used as an oriental medicinal herb to treat diabetes, diarrhea and various other inflammatory diseases. The phenolics of dry leaves of L. cuneata G. Don were extracted by using 80% (v/v) aqueous methanol in assistance with homogenization and sonification. The phenolic extract and its five different fractions (n-hexane, chloroform, ethyl acetate, n-butanol, and water) were used to evaluate the levels of total phenolics, total flavonoids, and antioxidant capacity as well as the inhibitory effect of tyrosinase activity. Ethyl acetate fraction (1 g) had the highest levels of total phenolics at 240.8 mg gallic acid equivalents (GAE), total flavonoids as 90.4 mg catechin equivalents (CE) as well as antioxidant capacity at 523.4 mg vitamin C equivalents (VCE) on ABTS assay and 329.5 mg VCE on DPPH assay among fractions. One g of water fraction contained total phenolics at 133.1 mg GAE, total flavonoids at 34.5 mg CE, and antioxidant capacity at 333.4 mg VCE for ABTS assay and 313.2 mg VCE for DPPH assay. Inhibition of tyrosinase activity of water fraction at 300 ${\mu}g{\cdot}mL^{-1}$ was at 47.2% and 21.1% for L-tyrosine and L-DOPA as its substrate, respectively. On the other hand, ethyl acetate fraction at 300 ${\mu}g{\cdot}mL^{-1}$ showed tyrosinase inhibition of 10.2% for L-tyrosine and 11.9% for L-DOPA. These results suggested that the phenolics from dry leaves of L. cuneata G. Don may be utilized as a potent source of antioxidants and skin whitening agents.

• Applied Chemistry for Engineering
• /
• v.28 no.2
• /
• pp.198-205
• /
• 2017

In this study, graviola leaf extracts (GLE) was investigated for the effect of antioxidant, antibacterial, whitening, anti-wrinkle. The antioxidant effect of GLE was measured by an electron donating ability assay. As a result, GLE increased the electron donating ability in a concentration-dependent manner. The antibacterial effect of GLE was found to show the higher antibacterial effect in Methicillin-resistant Staphylococcus aureus CCARM3115 compared with that of ampicillin by a paper disc method. The whitening effect of GLE was also measured by tyrosinase inhibition assay, and it was found that the tyrosinase activity of GLE decreased as the concentration increased. The inhibition activity of tyrosinase involved in hydroxylation reaction which is related to converting L-tyrosine to (DOPA) was higher than that of arbutin's at the concentration ranging from 125 to $250{\mu}g/mL$. In addition, GLE reduced melanin contents of B16-F10 melanoma cells in a dose-dependant manner and decreased to about 76.7% at a concentration of $100{\mu}g/mL$ Regarding wrinkling formation of GLE, an elastase inhibition assay was performed. As a result, GLE and ursolic acid were 10.5% and 56.5%, respectively under the identical concentration. These results suggest that GLE has significant antioxidant and whitening activities, and also may be potentially used as a therapeutic agent for hyperpigmentation treatment as an ingredient of whitening cosmetics.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.53 no.2
• /
• pp.187-195
• /
• 2008

Polyphenol oxidase (PPO) is implicated in discoloration of white salted noodles and other wheat based foods. PPO activity was evaluated to determine the effect on discoloration of noodle dough sheets prepared from 25 Korean wheat flours during storage and to screen experimental lines with low PPO activity in 52 Korean wheats. PPO activity was assayed with whole-seed and performed with L-dihydroxyphenylalanine (L-DOPA) as substrates. Absorbance by L-DOPA assay of 25 Korean wheats was from 0.285 to 1.368 at 475 nm. PPO activity was significantly related with grain characteristics, including 1000-kernel weight and grain colors. In flour characteristics, PPO activity positively correlated with ash and protein content (r = 0.658, P < 0.001 and r = 0.424, P < 0.05, respectively) and negatively correlated with $L^*$ value of flour (r = 0.412, P < 0.05). In the changes of color of noodle dough sheet, $L^*$ and $b^*$ values consistently decreased and $a^*$ value increased during storage. PPO activity negatively correlated with $L^*$ value of noodle dough sheet during storage (r = 0.566, P < 0.01 at 2 hr, r = 0.547, P < 0.01 at 24 hr, and r = 0.509, P < 0.01 at 48 hr). But, no significant relationship was found in between PPO activity, $a^*$ and $b^*$ values during storage. The 52 Korean wheat lines examined in this study were divided into 3 different groups, low (< 0.500), medium (0.501-0.999) and high level (> 1.000), on the basis of the level of PPO activity. Twenty two Korean wheat lines showed low level of PPO activity and Suwon 252, 277 and 280 showed lower PPO activity (< 0.200) than others.