• Journal of Applied Biological Chemistry
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• v.58 no.4
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• pp.295-301
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• 2015

Five phenylpropanoids (1-5), a benzofuran neolignan (6), two 8-O-4'-neolignans (7-8), and five tetrahydrofuran lignans (9-13) were isolated from a methanol extract of Myristica fragrans seeds. The structures of 1-13 were determined by $^1H$- and $^{13}C$-NMR spectroscopic data analyses and a comparison with the literature data. Compound 3 was isolated for the first time from this plant. All the isolated compounds were evaluated for their inhibitory activity against tyrosinase. Among them, safrole (1) showed significant inhibitions against both the monophenolase ($IC_{50}=32.11{\mu}M$) and diphenolase ($IC_{50}=27.32{\mu}M$) activities of tyrosinase. The kinetic analysis shows that safrole (1) is competitive inhibitors for both monophenolase and diphenolase. The apparent inhibition constant ($K_i$) for safrole (1) binding with free enzyme was determined to be 16.05 and $13.66{\mu}M$ for monophenolase and diphenolase, respectively.

• Biomolecules & Therapeutics
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• v.29 no.4
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• pp.445-451
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• 2021

Salicylamide, a non-steroidal anti-inflammatory drug (NSAID), is used as an analgesic and antipyretic agent. We have previously shown that several NSAIDs have anti-melanogenic properties in B16F1 melanoma cells. In contrast, we have found that salicylamide enhances melanin contents in B16F1 melanoma cells; however, the underlying mechanism is not known. Therefore, we investigated the mechanism through which salicylamide stimulates melanogenesis. Interestingly, salicylamide enhanced diphenolase activity in a cell-free assay. Western blotting and real-time RT-PCR revealed that salicylamide increased tyrosinase expression via transcriptional activation of the Mitf gene. Together, our results indicate that salicylamide could be used as an anti-hypopigmentation agent for skin and/or hair.

• Clean Technology
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• v.22 no.3
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• pp.175-180
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• 2016

Tyrosinases catalyze the hydroxylation of a monophenol (monophenolase activity) and the conversion of an o-diphenol to o-quinone (diphenolase activity), which are mainly involved in the modification of tyrosine residues into 3,4-dihydroxyphenyl-alanine (DOPA) and DOPA/DOPAquinone-derived intermolecular cross-linking. Previously, we obtained a slightly acidic and cold-active tyrosinase, tyrosinase-CNK, by our recombinant protein approach. The enzyme showed optimal activity at pH 6.0 and 20 ℃ with an abnormally high monophenolase/diphenolase activity ratio and still had approximately 50% activity compared with the highest activity even in ice water. Here, we investigated reaction stability of the recombinant tyrosinase-CNK as a psychrophilic enzyme. The enzyme showed remarkable thermal stability at 0 ℃ and the activity was well conserved in repeated freeze-thaw cycles. Although water-miscible organic solvent as reaction media caused the activity decrease of tyrosinase-CNK as expected, the enzyme activity was not additionally decreased with increased concentration in organic solvents such as ethanol and acetonitrile. Also, the enzyme showed high salt tolerance in chaotropic salts. It was remarkably considered that 2⁺ metal ions might inhibit the incorporation of Cu²⁺ into the active site. We expect that these results could be used to design tyrosinase-mediated enzymatic reaction at low temperature for the production of catechols through minimizing unwanted self-oxidation and enzyme inactivation.

• KSBB Journal
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• v.29 no.1
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• pp.1-8
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• 2014

Tyrosinases catalyze the hydroxylation of monophenolic compounds and the conversion of o-diphenols to oquinones. The enzymes are mainly involved in the modification of tyrosine into L-3,4-dihydroxyphenyl-alanine (L-DOPA) and DOPA/DOPAquinone-drived intermolecular cross-linking, which play the key roles of pigmentation to the cells. It is ubiquitously distributed in microorganisms, plants, and animals all around the nature world. They are classified as copper- containing dioxygen activating enzymes; two copper ions are coordinated with six histidine residues in their active sites and they are distinguished as met-, deoxy-, and oxy-form depending on their oxidative states. Natural extraction and recombinant protein approaches have been tried to obtain practical amounts of the enzymes for industrial application. Tyrosinases have been widely applied to industrial and biomedical usages such as detoxification of waste water containing phenolic compounds, L-DOPA as a drug of Parkinson's disease, biomaterials preparation based on the cross-linking ability and biosensors for the detection of phenolic compounds. Therefore, this review reports the mechanism of tyrosinase, biochemical and structural features and potential applications in industrial field.

• Journal of Microbiology and Biotechnology
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• v.32 no.5
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• pp.594-601
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• 2022

This study evaluated the biological properties of lemongrass (Cymbopogon citratus) extracts. The EtOAc extract of lemongrass had DPPH, TEAC, and nitric oxide-scavenging activity assay results of 58.06, 44.14, and 41.08% at the concentration of 50, 10, and 50 ㎍/ml, respectively. The EtOAc extract had higher elastase and collagenase inhibitory activities than the 80% MeOH, n-hexane, BuOH, and water extracts and comparable whitening activity toward monophenolase or diphenolase. Also, the EtOAc fraction had higher lipase inhibitory and antimicrobial activities against Cutibacterium acnes among extracts which is known to an important contributor to the progression of inflammatory acne vulgaris, and an opportunistic pathogen present in human skin. Total phenolic and flavonoid concentrations in the EtOAc extract were 132.31 mg CAE/g extract and 104.50 mg NE/g extract, respectively. Biologically active compounds in lemongrass extracts were analyzed by LC-MS. This study confirms that lemongrass extracts have potential use as cosmetic skincare ingredients. Thus, lemongrass can be considered a promising natural source of readily available, low-cost extracts rich in antioxidant, skincare, and antimicrobial compounds that might be suitable for replacing synthetic compounds in the cosmeceutical industry.

• The Korean Journal of Food And Nutrition
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• v.32 no.6
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• pp.611-619
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• 2019

The purpose of this study was to investigate the antioxidant activity and tyrosinase inhibitory activity of Codonopsis lanceolata 50% ethanol extract, and its solvent fractions (n-hexane, ethyl acetate (EA), n-butanol, water). The main components of the EA fraction were qualitatively analyzed using UPLC Q-ToF/MS. Additionally, a quantitative analysis was performed using UPLC. As a result, the total polyphenol content was 113.36 mg gallic acid/g in the EA fraction, which contained the largest amount of the C. lancolata solvent fractions. Also EA showed the highest antioxidant activity than other fractions. The IC₅₀ of DPPH(2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity was 0.03 mg/mL and the IC₅₀ of ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate)] radical scavenging activity was 0.049 mg/mL. The EA fraction showed tyrosinase inhibitory activity than other fractions and especially inhibited monophenolase oxidase reaction higher than diphenolase oxidase reaction. The monophenolase oxidase inhibited 55% when the concentration of the EA fraction was 0.25 mg/mL. As a result of Q-ToF/MS analysis, it was confirmed that tangshenoside I and lobetyolin were the main components of EA fraction. Thus, these results suggest that C. lanceolata may be used as a potent source of cosmetic agents.

• Journal of the Korean Society of Food Science and Nutrition
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• v.17 no.4
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• pp.348-357
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• 1988

The present work was undertaken to investigated the purification and characterization of polyphenol oxidase (PPO ; EC 1.10.3.1) in sweet potato, particularly the number of PPO isozymes, and PPO properties such as pH optimum, heat stability, substrate specificity, kinetics, and inhibitor studies. The purification achieved was 23.1 fold from crude extract with a yield of 41.5%. Eight PPO isozymes and twelve PPO isozymes were detected by disc polyacrylamide gel electrophoresis and isoelectric focusing, respectively. The specific activity of each isozyme separated by isoelectric focusing was in the range of $6,000{\sim}46,700U/mg$. This enzyme was sweet below $65^{\circ}C$ and the pH optimum of PPO occurred at 6.0-6.5. The substrate specificity of sweet potato PPO showed the high affinity toward the odiphenolic compounds. Km and Vmax for catechol were found to be 6.7 mM and $20{\triangle}A/min$, me protein, respectively. Inhibitor studies indicated that dithiothreitol was the most potent among the inhibitors used in the present work.

• Journal of Life Science
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• v.7 no.3
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• pp.192-197
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• 1997

Italian ryegrass, red clover, sorghum, and alfalfa were used for leaf protein preparation. Fresh leaves were pulped in the presence or absence of a reducing agent(sodium ascorbate or NaHSO$_{4}$) and green juice was heated and washed with acetone. The biological evaluation of leaf proteins was carried out by the growth method with male rats weighing about 45g. Italian ryegrass, red clover, and sorghum were brown when leaves were pulped in the absence of a reducing agent. On the other hand, alfalfa had neither o-diphenolics nor polyphenolase, and hence the alfalfa leaf protein did not brown during pulping ever in the absence of a reducing agent. The brown leaf protein from Italian ryegrass hd lower digestibility than the leaf protein protected from browning, although there were no difference in growth-promoting effect and protein efficiency ratio(PER)between the two leaf protein. The feeding of brown leaf protein from red clover resulted in the lowering of weight gain, digestibility, and PER, and all the measurement including diet intake were lowered by feeding the brown leaf protein from sorghum. In the case of alfalfa leaf protein, there were no difference in nutritional quality between the two leaf protein made with and without an attempt to prevent browning. The results mentioned above indicate that the occurrence of phenolics and polyphemolase in a crop in responsible for the browning of leaf protein and that the browning of leaf protein caused its nutritional impairment.