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

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.1
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• pp.29-33
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• 2004

Low molecular weight silk fibroin (LMSF), which was prepared by hydrolysis of silk fibroin using high-temperature and high-pressure method, was found to inhibit the oxidation of L-3,4,-dihydroxyphenylalanine (L-DOPA) catalyzed by mushroom tyrosinase (EC 1.14.18.1). LMSF contained mostly free amino acids such as L-glycine, L-alanine, and L-serine and oligopeptides, mainly glycine-alanine dimer. As a result of analyzing the inhibition kinetics from Lineweaver-Burk plots, L-glycine and glycine-alanine dimer showed noncompetitive behavior while uncompetitive behavior was observed in L-alanine, and L-serine. When weight percent concentration of ${ID_50}$ was compared, L-glycine was most effective on the inhibition and LMSF was also good enough for the inhibition effect of tyrosinase activity. LMSF showed a mixed-type inhibition and the inhibitory mechanism of LMSF might be caused by free amino acids and oligopeptides. As a result of spectroscopic observation with time, initial rate of increase of DOPAchrome decreased remarkably and the time to reach maximum absorbance increased as an increase of the concentration of L-glycine, meaning that L-glycine made itself mainly responsible for the formation of chelate with ${Cu^2+}$ in tyrosinase. However, in case of L-alanine, L-serine, and especially glycine-alanine dimmer, the production of DOPAchrome after an arrival at maximum absorbance decreased, indicating the production of adducts through the reaction with DOPAquinone.

• 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.

• Fisheries and Aquatic Sciences
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• v.25 no.6
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• pp.327-334
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• 2022

Marine resources have various biological activities and their constituents are more novel than those of land organisms. Several biologically active constituents have been found in marine organisms. Recently, many studies have reported that marine animals (MAs) can be used as functional ingredients in functional foods or nutraceutical due to their health benefits. However, no studies have extensively investigated the cosmeceutical activities of MAs extracts. Here, 70% ethanol extracts of 38 MAs were investigated for their activities of whitening and anti-aging properties for use as materials in novel cosmeceuticals. Anti-aging activities were determined by skin aging-related enzyme activities (anti-collagenase, anti-elastase, anti-hyaluronidase) and whitening activities (anti-tyrosinase, anti-3,4-dihydroxyl-L-phenylalanine [DOPA] oxidation) evaluated by colorimetric method. Among the 38 MAs, we found that Urechis unicinctus and Petrosia corticata extracts showed the strongest inhibitory effects against tyrosinase and DOPA oxidation, respectively. Our results additionally showed that Protankyra bidentata extract might provide a major source of anti-hyaluronidase and anti-elastase; meanwhile, anti-collagenase effects were similar in most MAs. Overall, these results suggest that extracts of marine animals have potential as a tyrosinase, collagenase, elastase, and hyaluronidase inhibitors. Taken together, MA resources could be considered as a novel cosmeceutical agent to be applied in cosmetic industry.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1513-1520
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• 2010

A bacterium capable of producing melanin pigment in the presence of L-tyrosine was isolated from a crop field soil sample and identified as Klebsiella sp. GSK based on morphological, biochemical, and 16S rDNA sequencing. The polymerization of this pigment occurs outside the cell wall, which has a granular structure as melanin ghosts. Chemical characterization of the pigment particles showed then to be acid resistant, alkali soluble, and insoluble in most of the organic solvents and water. The pigment got bleached when subjected to the action of oxidants as well as reductants. This pigment was precipitated with $FeCl_3$, ammoniacal silver nitrate, and potassium ferricynide. The pigment showed high absorbance in the UV region and decreased absorbance when shifted towards the visible region. The melanin pigment was further charecterized by FT-IR and EPR spectroscopies. A key enzyme, 4-hydroxyphenylacetic acid hydroxylase, that catalyzes the formation of melanin pigment by hydroxylation of L-tyrosine was detected in this bacterium. Inhibition studies with specific inhibitors, kojic acid and KCN, proved that melanin is synthesized by the DOPA-melanin pathway.

• Proceedings of the SCSK Conference
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• 2003.09b
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• pp.241-242
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• 2003

The effects of N-acetylphytospingosine(NAPS), one of the phytospingosine derivatives, on melanogenesis of B 16F 1 0 mouse melanoma cell lines were investigated. We assessed the effect of NAPS on the depigmentation of B16F10 cells. The melanin content of cells was significantly reduced by NAPS. We examined the inhibitory effect of NAPS on tyrosinase activity using L-dopa as a substrate and the results showed that tyrosinase activity was inhibited in a does-dependent manner. The mRNA level of tyrosinase as well as that of tyrosinase related protein-l (TRP-l) and tyrosinase related protein-2 (TRP-2) genes were not affected by NAPS based on a reverse transcription-polymerase chain reaction (RT-PCR) assay. We also performed a Western blotting analysis using anti-tyrosinase antibody. It showed that there is no change in tyrosinase protein level after treatment of NAPS. These results suggest that the depigmenting mechanism of NAPS in B16F10 melanoma cells involves inhibition of melanosomal tyrosinase activity, rather than the mRNA expression or protein level of tyrosinase.

• Molecules and Cells
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• v.45 no.9
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• pp.640-648
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• 2022

CD133, also known as prominin-1, was first identified as a biomarker of mammalian cancer and neural stem cells. Previous studies have shown that the prominin-like (promL) gene, an orthologue of mammalian CD133 in Drosophila, plays a role in glucose and lipid metabolism, body growth, and longevity. Because locomotion is required for food sourcing and ultimately the regulation of metabolism, we examined the function of promL in Drosophila locomotion. Both promL mutants and pan-neuronal promL inhibition flies displayed reduced spontaneous locomotor activity. As dopamine is known to modulate locomotion, we also examined the effects of promL inhibition on the dopamine concentration and mRNA expression levels of tyrosine hydroxylase (TH) and DOPA decarboxylase (Ddc), the enzymes responsible for dopamine biosynthesis, in the heads of flies. Compared with those in control flies, the levels of dopamine and the mRNAs encoding TH and Ddc were lower in promL mutant and pan-neuronal promL inhibition flies. In addition, an immunostaining analysis revealed that, compared with control flies, promL mutant and pan-neuronal promL inhibition flies had lower levels of the TH protein in protocerebral anterior medial (PAM) neurons, a subset of dopaminergic neurons. Inhibition of promL in these PAM neurons reduced the locomotor activity of the flies. Overall, these findings indicate that promL expressed in PAM dopaminergic neurons regulates locomotion by controlling dopamine synthesis in Drosophila.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.4
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• pp.287-301
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• 2008

In order to develop a new depigmenting agent, extracts were obtained from 60 native plants and their antimelanogenic activities were screened by evaluating the inhibitory effect on tyrosinase which is a major enzyme responsibles for the melanin synthesis. The extracts of Trichosanthes kirilowii fruits, Phyllostachys bambusoides inner films (BIF), Clerodendrum trichotomum leaves, and Acer okamotoanum leaves showed relatively high inhibitory effect on tyrosinase and their $IC_{50}$ values were $50{\sim}100{\mu}g/mL$. The extract of BIF inhibited melanin synthesis of B16F10 melanoma cells by 52%, which was the highest among those of various extracts. Furthermore, the effect of BIF extract is 10% higher than that of arbutin (42%), a popular depigmenting agent in Korea. Ten compounds having antimelanogenic activity were isolated from the BIF extract by solvent extraction and chromatography. These compounds were identified as phenolic derivatives: SM701, SM702, SM703, and BPR211 were hydroquinone derivatives; SM707 a gallic acid derivative; SM704, SM705, SM706, SM708 and SM709 ferulic acid derivatives. The free radical scavenging activities of these compounds were measured and compared to those of hydroquinone and vitamin C. The $SC_{50}$ values scavenging 50% DPPH of SM702 and SM709 were $60{\sim}70{\mu}M$ similar to that of hydroquinone and those of SM701 and SM708 were $30{\sim}40{\mu}M$ slightly lower than that of vitamin C. These results suggest the presence of components having high antioxidant activity in the BIF extract. The SM709, identified as 1,2-O-diferulylglycerol, inhibited the activities of tyrosine hydroxylase and dopa oxidase by 18 and 60%, respectively. The SM709 also inhibited the melanin synthesis of B16F10 melanoma cells by 62% and this was the highest antimelanogenic activity among those obtained from the various purified compounds. Therefore, antimelanogenic activity of the BIF extract was concluded to be due to both inhibition of DOPA oxidase and antioxidant activity.

• Journal of Life Science
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• v.22 no.7
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• pp.936-942
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• 2012

Prunus mume has been traditionally used as a medicinal food in Korea, Japan, and China. In particular, this fruit has been reported to have beneficial biological effects on gastritis and gastric ulcers. However, its action in relation to skin whitening has remained unclear. Accordingly, the effects of fruit extract of P. mume related to antioxidation and skin whitening were examined in this study. First, using the MTT assay, it was observed that fruit extract of P. mume below 0.1% has no cytotoxicity in B16-F1 cells as a result of cell viability. Second, the direct scavenging effects and the reducing power of the fruit extract of P. mume were evaluated in vitro on DPPH radicals, hydrogen peroxide, and superoxide. It exhibited high reducing power and scavenging activity on the aforementioned reactive oxygen species. Furthermore, we found that its protective effect against genomic DNA damage related to oxidative stress was increased in a dose-dependent manner. In addition, the fruit extract of P. mume had an inhibitory effect on melanin production induced by L-dopa. In addition, it reduced the expression level of NRF-2, SOD-1, and SOD-2 related to antioxidation in western blot analysis. These results suggest that fruit extract of P. mume could exert a whitening effect through inhibition of melanin production by its antioxidant effect.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.207-214
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• 2012

Laccase- and peroxidase-free tyrosinase has commercial importance in the production of L-3, 4-dihydroxyphenylalanine (L-DOPA), which is mainly used in the treatment of Parkinson's disease. In the present study, isolation of an actinomycetes microbial strain capable of producing only tyrosinase is reported. Among all soil isolates, three individual colonies revealed black color around the colony in the presence of tyrosine. Further screening for laccase and peroxidase activities using syringaldazine denoted that one of the isolates, designated as RSP-T1, is laccase and peroxidase negative and produces only tyrosinase. The microbe was authenticated as Streptomyces antibioticus based on 16S ribotyping. Effective growth of this isolate was noticed with the use of medium (pH 5.5) containing casein acid hydrolysate (10.0 g/l), $K_2HPO_4$ (5.0 g/l), $MgSO_4$ (0.25 g/l), L-tyrosine (1.0 g/l), and agar (15 g/l). The scanning electron micrograph depicted that the microbe is highly branched and filamentous in nature. The enzyme production was positively regulated in the presence of copper sulfate. The impact of different fermentation parameters on tyrosinase production depicted that the maximized enzyme titer values were observed when this isolate was grown at 6.5 pH and at $30^{\circ}C$ temperature under agitated conditions (220 rpm). Among all the studied physiological parameters, agitation played a significant role on tyrosinase production. Upon optimization of the parameters, the yield of tyrosinase was improved more than 100% compared with the initial yield.