• Title/Summary/Keyword: Tyrosinase Gene

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Microbial Strains and Bioactive Exopolysaccharide Producers from Thai Water Kefir

  • Luang-In, Vijitra;Saengha, Worachot;Yotchaisarn, Manatchanok;Halaslova, Monika;Udomwong, Piyachat;Deeseenthum, Sirirat
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.403-415
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    • 2018
  • The aims of this novel work were to determine the microbial strains and exopolysaccharide (EPS) producers in water kefir from Nakhon Ratchasima Province, Thailand. Thirty-three microbial strains were identified using 16S rRNA gene analysis consisting of 18 bacterial strains, as 9 strains of acetic acid bacteria (AAB), 9 strains of lactic acid bacteria (LAB), and 15 yeast strains. All bacteria were able to produce EPS with a diverse appearance on agar media containing different sugars at a concentration of 8%. Culture supernatants from AAB and LAB showed 31-64% 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity with the highest antioxidant activity of 64% from Acetobacter pasteurianus WS3 and WS6. Crude EPS from A. pasteurianus WS3 displayed the highest ferric reducing antioxidant power at 280 mM $FeSO_4/g$ EPS, greatest anti-tyrosinase activity at 20.35%, and highest EPS production of 1,505 mg EPS/L from 8% sucrose. These microbes offer beneficial health implications and their EPSs can be used as food additives and cosmetic ingredients.

Characterization of Recombinant Derivatives of pJY711 of Multicopy Streptomyces Plasmid (Multicopy Streptomyces 플라스미드 pJY711의 재조합 유도체의 특성)

  • 염도영;공인수;유주현
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.35-40
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    • 1990
  • The restriction clevage map of multi-copy recombinant plasmid, pJY712(8.1kb), carrying the thiostrepton resistance gene(tsr) was determined. pJY712 had a broad host range in Streptomyces and contained single BglII site for cloning purpose. The plasmid showed the phenomenon of lethal zygosis ($Ltz^{+}$). Transformation frequency of pJY712 was $5.0\times 10^{4}$ transformants per ug plasmid DNA (TFU) in S. lividans. Plasmid pJY713 was constructed by inserting the tyrosinase gene(mel) into the BclI site of pJY712. Recombinant plasmid pJY714 carrying the mel gene was constructed by in vitro deletion of a segment (1.9kb BglII-BclI fragment) from pJY713.

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Mutations of TYR and MITF Genes are Associated with Plumage Colour Phenotypes in Geese

  • Wang, Ye;Li, Si-Ming;Huang, Jing;Chen, Shi-Yi;Liu, Yi-Ping
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.6
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    • pp.778-783
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    • 2014
  • The polymorphism of microphthalmia-associated transcription factor (MITF) and tyrosinase (TYR) genes have been proposed to play a vital role in coat colour genesis in mammals, but their role remains ambiguous in geese at best. Here, we cloned and sequenced 1,397 bp coding region of MITF gene and a 588 bp fragment of TYR exon 1 for polymorphism analysis among 157 domestic geese showing three types of plumage colour. We detected a total of three SNPs (c.280T>C, c.345G>A, and c.369G>A) in TYR and six haplotypes (H1-H6). Among them, haplotypes H1, H2, H3, and H5 were significantly associated with white plumage trait of Zhedong White Geese. However, only diplotype H1H1 and H3H5 were significantly associated with white plumage trait of Zhedong White Geese (p<0.01). We only detected one SNP (c.1109C>T) for MITF gene and found that genotype CT and TT were significantly associated with white plumage trait of Zhedong White Geese. Briefly, our study suggested an association between polymorphisms of TYR and MITF genes and the plumage colour trait in domestic geese.

Genetic Variations of Chicken TYR Gene and Associations with Feather Color of Korean Native Chicken (KNC) (한국 토종닭 모색 변이와 TYR 유전자형 간의 상관관계 분석)

  • Choi, Jin Ae;Lee, Jun-Heon;Jang, Hyun-Jun;Lee, Kyung-Tai;Kim, Tae-Hun;Lee, Hyun-Jeong;Heo, Kang-Nyeong;Kim, Chong-Dae;Han, Jae-Yong;Park, Mi Na
    • Korean Journal of Poultry Science
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    • v.41 no.1
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    • pp.7-14
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    • 2014
  • Tyrosinase (TYR) gene is located on chromosome 1 in chicken and it is composed of five exons and four introns. TYR gene is described as a key enzyme in melanin biosynthesis. Most examples of complete albinism in chicken have been due to defects in the tyrosinase gene. The association of feather color and sequence polymorphism in the Tyrosinase (TYR) gene was investigated using Korean Native chicken H breed (H_PL), Korean Native chicken L/W breed(L/W_PL) and 'Woorimatdag' commercial chickens (Woorimatdag_CC). From L_PL and W_PL breed analyses, 4 synonymous SNPs (locus G33A, G116A, C217T and C247T) and 2 SNPs (G838A and G958A) were detected in 4th exon and 4th intron of TYR gene respectively. The genotype frequencies for 6 SNPs were compared between L_PL and W_PL and W_PL represented homozygous SNP types in all the analyzed SNP positions while L_PL displayed various SNP types.

Identification of a Gene Involved in the Negative Regulation of Pyomelanin Production in Ralstonia solanacearum

  • Ahmad, Shabir;Lee, Seung Yeup;Khan, Raees;Kong, Hyun Gi;Son, Geun Ju;Roy, Nazish;Choi, Kihyuck;Lee, Seon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1692-1700
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    • 2017
  • Ralstonia solanacearum causes bacterial wilt in a wide variety of host plant species and produces a melanin-like blackish-brown pigment in stationary phase when grown in minimal medium supplemented with tyrosine. To study melanin production regulation in R. solanacearum, five mutants exhibiting overproduction of melanin-like pigments were selected from a transposon (Tn) insertion mutant library of R. solanacearum SL341. Most of the mutants, except one (SL341T), were not complemented by the original gene or overproduced melanins. SL341T showed Tn insertion in a gene containing a conserved domain of eukaryotic transcription factor. The gene was annotated as a hypothetical protein, given its weak similarity to any known proteins. Upon complementation with its original gene, the mutant strains reverted to their wild-type phenotype. SL341T produced 3-folds more melanin at 72 h post-incubation compared with wild-type SL341 when grown in minimal medium supplemented with tyrosine. The chemical analysis of SL341T cultural filtrate revealed the accumulation of a higher amount of homogentisate, a major precursor of pyomelanin, and a lower amount of dihydroxyphenylalanine, an intermediate of eumelanin, compared with SL341. The expression study showed a relatively higher expression of hppD (encoding hydroxyphenylpyruvate dioxygenase) and lower expression of hmgA (encoding homogentisate dioxygenase) and nagL (encoding maleylacetoacetate isomerase) in SL341T than in SL341. SL341 showed a significantly higher expression of tyrosinase gene compared with SL341T at 48 h post-incubation. These results indicated that R. solanacearum produced both pyomelanin and eumelanin, and the novel hypothetical protein is involved in the negative regulation of melanin production.

Rare ginsenoside Ia synthesized from F1 by cloning and overexpression of the UDP-glycosyltransferase gene from Bacillus subtilis: synthesis, characterization, and in vitro melanogenesis inhibition activity in BL6B16 cells

  • Wang, Dan-Dan;Jin, Yan;Wang, Chao;Kim, Yeon-Ju;Perez, Zuly Elizabeth Jimenez;Baek, Nam In;Mathiyalagan, Ramya;Markus, Josua;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.42-49
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    • 2018
  • Background: Ginsenoside F1 has been described to possess skin-whitening effects on humans. We aimed to synthesize a new ginsenoside derivative from F1 and investigate its cytotoxicity and melanogenesis inhibitory activity in B16BL6 cells using recombinant glycosyltransferase enzyme. Glycosylation has the advantage of synthesizing rare chemical compounds from common compounds with great ease. Methods: UDP-glycosyltransferase (BSGT1) gene from Bacillus subtilis was selected for cloning. The recombinant glycosyltransferase enzyme was purified, characterized, and utilized to enzymatically transform F1 into its derivative. The new product was characterized by NMR techniques and evaluated by MTT, melanin count, and tyrosinase inhibition assay. Results: The new derivative was identified as (20S)-$3{\beta},6{\alpha},12{\beta}$,20-tetrahydroxydammar-24-ene-20-O-${\beta}$-D-glucopyranosyl-3-O-${\beta}$-D-glucopyranoside(ginsenoside Ia), which possesses an additional glucose linked into the C-3 position of substrate F1. Ia had been previously reported; however, no in vitro biological activity was further examined. This study focused on the mass production of arduous ginsenoside Ia from accessible F1 and its inhibitory effect of melanogenesis in B16BL6 cells. Ia showed greater inhibition of melanin and tyrosinase at $100{\mu}mol/L$ than F1 and arbutin. These results suggested that Ia decreased cellular melanin synthesis in B16BL6 cells through downregulation of tyrosinase activity. Conclusion: To our knowledge, this is the first study to report on the mass production of rare ginsenoside Ia from F1 using recombinant UDP-glycosyltransferase isolated from B. subtillis and its superior melanogenesis inhibitory activity in B16BL6 cells as compared to its precursor. In brief, ginsenoside Ia can be applied for further study in cosmetics.

Antioxidant effects and tyrosinase and elastase inhibitory activities of mountain ginseng adventitious roots extracts at different ethanol concentrations

  • Ko, Hyun Min;Eom, Tae Kil;Kim, Kyeoung Cheol;Kim, Chul Joong;Lee, Jae Geun;Kim, Ju-Sung
    • Korean Journal of Agricultural Science
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    • v.45 no.3
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    • pp.499-508
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    • 2018
  • To use mountain ginseng adventitious roots (MGAR), which is superior in functionality compared to ginseng, and to mass produce raw materials as a functional cosmetic material, the antioxidant and cosmeceutical activities of the extracts of MGAR at different ethanol concentrations (0, 20, 40, 60 and 80%) were compared and analyzed. At first, different ethanol concentrations were compared to determine the best solvent for the extraction of phenolic compounds from MGAR. An 80% ethanol extract with 10.07 mg of gallic acid equivalents/g sample and 0.94 mg of quercetin equivalents/g sample was the best solvent in the extraction of phenolic compounds. However, the 60% ethanol extract had the highest antioxidant activity, evident by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, Trolox equivalent antioxidant capacity (35.01 mM Trolox/g sample) and ferric reducing antioxidant power (460.94 mM $FeSO_4/g$ sample). However, the inhibitory activities of the 80% ethanol extract from the MGAR against tyrosinase and elastase were higher than those of the other extracts. The results of this study show that the extract of MGAR can be used as an easily accessible source of natural antioxidant and as a possible cosmetic ingredient in the cosmeceutical industry. However, the components responsible for the antioxidant and cosmeceutical activities of the MGAR extracts are currently unclear. Therefore, it is suggested that further studies should be carried out to isolate and identify the antioxidant and cosmeceutical components in MGAR.

Effects of Thujae Orientalis Folium (TOF) on Gene Expression of Human melanoma cells (SK-MEL-2) (측백엽(側柏葉)이 인간 유래 악성 흑색종 세포의 유전자 발현에 미치는 영향)

  • Jung, Min-Young;Kim, Jong-Han;Park, Su-Yeon;Choi, Jeong-Hwa
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.2
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    • pp.81-108
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    • 2010
  • Objective : Thujae Orientalis Folium (TOF) can cool the blood and stop bleeding, eliminate phlegm and relieve cough in Oriental medicine. In addition, the fresh is used alone externally. Recently, TOF is known to have anti-tumor component. And also known to have tyrosinase inhibitory effect. Method : For these reasons, this study was designed to investigate anti-cancer and whitening activities of TOF. In this experiment, effects of TOF on proliferation rates of melanoma cells and on changes in genetic profiles were investigated. The genetic profile for the effect on human derived melanoma cell, SK-MEL-2, was measured using microarray technique, and the functional analysis on these genes was conducted. Results : Total 541 genes were up-regulated and 1,079 genes down-regulated in cells treated with TOF. Genes induced by TOF were mainly concerned with anti-cancer effects and apoptosis. Genes suppresed by TOF were related in extracellular signalling pathway. The network of total protein interactions was measured using cytoscape program, and some key molecules, such as THAP1, MAX1, STAM2, SMAD6, CYCS, PEX5, PSEN1, NONO, MAP2K7 and CREB1 that can be used for elucidation of therapeutical mechanism of medicine in future were identified. Conculusion : These results suggest possibility of TOF as anti-cancer drug for human melanoma. In addition, the present author also suggest that related mechanisms are involved in inhibition of several cancer pathway, activation of apoptosis pathway and suppression of general metabolic pathway.

Extracellular Vesicles from Korean Codium fragile and Sargassum fusiforme Negatively Regulate Melanin Synthesis

  • Jang, Bohee;Chung, Heesung;Jung, Hyejung;Song, Hyun-Kuk;Park, Eunhye;Choi, Hack Sun;Jung, Kyuhyun;Choe, Han;Yang, Sanghwa;Oh, Eok-Soo
    • Molecules and Cells
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    • v.44 no.10
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    • pp.736-745
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    • 2021
  • Although various marine ingredients have been exploited for the development of cosmetic products, no previous study has examined the potential of seaweed extracellular vesicles (EV) in such applications. Our results revealed that EV from Codium fragile and Sargassum fusiforme effectively decreased α-MSH-mediated melanin synthesis in MNT-1 human melanoma cells, associated with downregulation of MITF (microphthalmia-associated transcription factor), tyrosinase and TRP1 (tyrosinase-related proteins 1). The most effective inhibitory concentrations of EV were 250 ㎍/ml for S. fusiforme and 25 ㎍/ml for C. fragile, without affecting the viability of MNT-1 cells. Both EV reduced melanin synthesis in the epidermal basal layer of a three-dimensional model of human epidermis. Moreover, the application of the prototype cream containing C. fragile EV (final 5 ㎍/ml) yielded 1.31% improvement in skin brightness in a clinical trial. Together, these results suggest that EV from C. fragile and S. fusiforme reduce melanin synthesis and may be potential therapeutic and/or supplementary whitening agents.

Ethanol extract of medicinal herbal mixture accelerates hair growth and melanogenesis in vivo and in vitro (한약재 복합추출물이 모발 성장 및 멜라닌 생성 촉진에 미치는 영향)

  • Kim, Ji Yoon;Kim, Yoo-Jin;Kim, Moon Ju;Kim, Mi Ryeo
    • The Korea Journal of Herbology
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    • v.33 no.5
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    • pp.9-18
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    • 2018
  • Objectives : This study was performed to determine the transdermal effects of ethanol extract from medicinal herbal mixture (SHJ) on hair growth in C57BL/6 mice and melanogenesis in melanoma cells. Methods : Mice were divided into 3 experimental groups including vehicle (CON), SHJ extract and 5% minoxidil (MNXD, positive control)-treated group. SHJ was applied topically on the hair-shaved skin of C57BL/6 mice everyday for 15 days. The thickness and density of hair with a folliscope and morphometry of hair follicle with a H&E staining were monitored at last day. Also then, hair growth-associated gene expressions were measured by immunoblot assay. Results : The MNXD or SHJ-treated group promoted on hair growth compared to that of vehicle-treated group (CON). Hair density and thickness of MNXD or SHJ treated-group increased compared to that of vehicle application on the 15 days, respectively. Induction of insulin-like growth factor (IGF)-1 and vascular endothelial growth factor (VEGF) were also accelerated by application of SHJ extract compared to those of CON group. But expression of transforming growth factor (TGF)-${\beta}1$ decreased in SHJ treated-group compared to that of CON group. Furthermore, SHJ extract showed to increase melanin contents in a dose-dependent manner. Tyrosinase activity significantly increased in SHJ-treated group compared with CON group in dose-dependant manner. Conclusions : These results suggest that SHJ can be used as a component of cosmeceuticals for hair care via promoting growth and melanogenesis of hair.