• Title/Summary/Keyword: TRP

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Genetic Analysis of Recombinants by Interspecific Protoplast Fusion of Coryneform Bacteria and Their L-glutamate & L-glutamine Production (Corynebacterium 세균의 이종간 원형질체 융합에 의한 재조합주의 유전학적 분석과 L-glutamate와 L-glutamine 생성)

  • 백선영;이혜경;최순영;김종욱;이세배;임번삼;민경희
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.296-300
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    • 1990
  • For interspecific portoplast fusion, Brevibacterium flauum lOAHR (Rifr axg his) and Corynebacterium glutamicum 11TS ($Sm-r$ trp) were induced by UV and NTG treatment. The protoplast fusion frequency between E. flavum XOAHR and C. glutamicum llTS was $3.7\times 10^{-6}$ with the lysozyme treatment (300 P $\mu g$ml) for 18 hrs. Genotypes of recombinants were analized as FMM ($Rif^r\; Sm^r$), FA (Rift $Sm^r$ arg), FH ($Rif^r\; Sm^r$ his), FT ($Rif^r\; Sm^r$ trp), FAH ($Rif^r\; Sm^r$ arg trp), FAT ($Rif^r\; Sm^r$ arg trp), and FAHT ($Rif^r\; Sm^r$ arg his trp). FAH 1 produced 12 fold of glutamate production compared to parental type, E. flauum 10AHR. In glutamine productivity, it produced 2.6 fold to parental type, C. glutamicum 11TS. Production of glutamate or glutamine by recombinants was involved in the specific activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS), respectively.

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Involvement of Transglutaminase-2 in α-MSH-Induced Melanogenesis in SK-MEL-2 Human Melanoma Cells

  • Kim, Hyun Ji;Lee, Hye Ja;Park, Mi Kyung;Gang, Kyung Jin;Byun, Hyun Jung;Park, Jeong Ho;Kim, Mi Kyung;Kim, Soo Youl;Lee, Chang Hoon
    • Biomolecules & Therapeutics
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    • v.22 no.3
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    • pp.207-212
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    • 2014
  • Skin hyperpigmentation is one of the most common skin disorders caused by abnormal melanogenesis. The mechanism and key factors at play are not fully understood. Previous reports have indicated that cystamine (CTM) inhibits melanin synthesis, though its molecular mechanism in melanogenesis remains unclear. In the present study, we investigated the effect of CTM on melanin production using ELISA reader and the expression of proteins involved in melanogenesis by Western blotting, and examined the involvement of transglutaminase-2 (Tgase-2) in SK-MEL-2 human melanoma cells by gene silencing. In the results, CTM dose-dependently suppressed melanin production and dendrite extension in a-MSH-induced melanogenesis of SK-MEL-2 human melanoma cells. CTM also suppressed a-MSH-induced chemotactic migration as well as the expressions of melanogenesis factors TRP-1, TRP-2 and MITF in a-MSH-treated SK-MEL-2 cells. Meanwhile, gene silencing of Tgase-2 suppressed dendrite extension and the expressions of TRP-1 and TRP-2 in a-MSH-treated SK-MEL-2 cells. Overall, these findings suggested that CTM suppresses a-MSH-induced melanogenesis via Tgase-2 inhibition and that therefore, Tgase-2 might be a new target in hyperpigmentation disorder therapy.

Whitening Effect of Watersoluble Royal Jelly from South Korea

  • Han, Sang Mi;Kim, Jung Min;Hong, In Phyo;Woo, Soon Ok;Kim, Se Gun;Jang, Hye Ri;Park, Kwan Kyu;Pak, Sok Cheon
    • Food Science of Animal Resources
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    • v.35 no.5
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    • pp.707-713
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    • 2015
  • Royal jelly has been widely used as a health supplement worldwide. However, royal jelly has been implicated in allergic reactions, and we developed a water-soluble royal jelly (WSRJ) without the allergy inducing protein. In this study, we aimed to identify the anti-melanogenic efficacy of WSRJ. B16F1 melanoma cells were first treated with 10 nM α-melanocyte stimulating hormone (α-MSH) and then with various doses of WSRJ. In addition, we investigated the mRNA and protein expression of melanogenesis-related genes such as tyrosinase, tyrosinase related protein-1 (TRP-1) and TRP-2 by reverse transcription-polymerase chain reaction and western blotting. WSRJ directly inhibited tyrosinase and cellular tyrosinase activity, which decreased melanin synthesis in α-MSH stimulated B16F1 melanoma cells a level comparable to that observed with arbutin. WSRJ decreased the mRNA and protein expressions of tyrosinase, TRP-1, and TRP-2, which was comparable to that observed with arbutin. WSRJ has strong anti-melanogenic activity, which invoice direct inhibition of tyrosinase enzyme activity and suppression of expression of melanogenesis related genes. Results from this study suggests that WSRJ is a potential candidate for the treatment of skin pigmentation.

A Study on the Depigmenting Effect of Carthamus tinctorius Seed, Cyperus rotundus and Schizonepeta tenuifolia Extracts (홍화자, 향부자, 형개 추출물의 미백효과에 관한 연구)

  • Hwang, Eun-Young;Kim, Dong-Hee;Hwang, Jo-Young;Kim, Hui-Jeong;Park, Tae-Soon;Lee, In-Sun;Son, Jun-Ho
    • Korean Journal of Food Science and Technology
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    • v.44 no.1
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    • pp.76-81
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    • 2012
  • The objective of the present study was to evaluate the skin depigmentation effect of the extracts of three herbs, Carthamus tinctorius seed, Cyperus rotundus and Schizonepeta tenuifolia. Their effects on tyrosinase and melanin synthesis inhibitory action were assessed. We found that the C. tinctorius seed ethanol extracts reduced the tyrosinase activity and melanin formation of B16F10 melanoma cells. The C. tinctorius seed suppressed the expression in microphthalmia associated transcription factor (MITF), tyrosinase, tyrosinase related protein 1 (TRP-1), and tyrosinase related protein 2 (TRP-2) in B16F10 melanoma cells. These results show that C. tinctorius seed inhibited melanogenesis on the B16F10 melanoma cell. The underlying mechanism of C. tinctorius seed whitening activity may be the inhibition of tyrisinase, MITF, tyrosinase, TRP-1, and TRP-2 expression. The results suggested that C. tinctorius seed has considerable potential as a natural functional ingredient with a depigmentation effect.

The Studies on the Heterogenous Gene Expression in Schizophyllum commune (치마버섯에서 이형 유전자 발현에 관한 연구)

  • Park, Dong-Chul;Kim, Hyun-Jeong;Kim, Ok-Mi;Bae, Jun-Tae;Park, Sun-Hee;Lee, Byeung-Hun;Lee, Kap-Rang
    • The Korean Journal of Mycology
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    • v.26 no.1 s.84
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    • pp.103-107
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    • 1998
  • The trp2 gene encoding trifunctional enzyme in Coprinus cinereus was investigated the expression in the heterothallic mushroom species. To identify the homology of trp2 gene to Schizophyllum commune and Pleurotus ostreatus, southern hybridization was performed with plasmid pHIONA8 containing C. cinereus trp2 gene as a probe, which resulted in a strong signal indicating an homologous sequence to the chromosomal DNA of S. commune. About 50 transformants per dish was appeared in the complementation test by pHIONA8 using S. commune tryptophan auxotroph as host. In the mating test between transformants and other mating type alleles, the fruiting body of S. commune was formed at $30^{\circ}C$ in $2{\sim}3$ weeks.

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Capsosiphon Fulvescens Decreases Melanin Synthesis Via Downregulation of Tyrosinase and TRP-2 Expression

  • Yoo, Hyun-Ju;Jo, Mi-Gyeong;Park, Si-Jun;Mun, Yeun-Ja;Pyo, Hyeong-Bae;Lim, Kyu-Sang;Lee, Ki-Nam;Woo, Won-Hong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.4
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    • pp.998-1003
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    • 2007
  • The green marine algae, Capsosiphon fulvescens (CF) is a food supplement cultivated in south coast of Southern Korea. The purpose of this study was to investigate the mechanism of CF-induced hypopigmentation. The present study was designed to determine the effect of CF extracton melanogenesis in B16 cells, particularly its specific effects on tyrosinase and tyrosinase-related protein 2 (TRP-2). We measured melanin contents and analyzed melanosome associated protein levels using Western blot and Reverse transcription-polymerase chian reaction (RT-PCR) analysis. CF extract markedly inhibited melanin synthesis and tyrosinase activity. In addition, cellular dendricity was slightly decreased by CF extract. In further experiments, CF extract significantly reduced the protein levels of tyrosinase and TRP-2 in B16 cells. RT-PCR analysis revealed that tyrosinase and TRP-2 mRNA levels were unaffected by CF treatment. Therefore, these results suggest that hypopigmentary effect of CF was due to post-translational degradationof tyrosinase and TRP-2.

Inhibitory Effects on Melanin Production of Demethylsuberosin Isolated from Angelica gigas Nakai (참당귀로부터 분리한 Demethylsuberosin의 멜라닌 생성 억제 효과)

  • Kim, You Ah;Park, Sung Ha;Kim, Bo Yun;Kim, A Hyun;Park, Byoung Jun;Kim, Jin Jun
    • Korean Journal of Pharmacognosy
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    • v.45 no.3
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    • pp.209-213
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    • 2014
  • The anti-melanogenic substance was isolated from the root of Angelica gigas Nakai by silica gel column chromatography, preparative HPLC and TLC. As a result of the structure analysis by mass, $^1H$-NMR, and $^{13}C$-NMR spectrometry, the compound was identified as demethylsuberosin. Demethylsuberosin reduced melanin contents of B16F1 melanoma cells in a dose-dependent manner and decreased to about 74% at a concentration $5{\mu}g/ml$. Demethylsuberosin inhibited the expression in microphthalmia associated transcription factor (MITF), tyrosinase, tyrosinase related protein-1 (TRP-1), and tyrosinase related protein-2 (TRP-2) in melanocytes. These results suggest that the whitening activity of demethylsuberosin may be due to the inhibition of the melanin synthesis by down-regulation of MITF, tyrosinase, TRP-1 and TRP-2 expression. Thus, our results provide evidence that demethylsuberosin might be useful as a potential skin-whitening agent.

Inhibitory effects of crude polysaccharide fractions from Annona muricata L. on melanogenesis (그라비올라 잎(Annona muricata L.) 조다당 분획분의 멜라닌 생성 저해 효과)

  • Kim, Yi-Eun;Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.51 no.1
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    • pp.52-57
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    • 2019
  • The objective of this study was to evaluate the anti-melanogenic effects of crude polysaccharide fractions from Annona muricata L. (ALP) in 3-isobutyl-1-methylxanthine (IBMX) stimulating hormone-induced mouse B16F10 melanoma cells. The inhibitory effect of ALP on tyrosinase activity was approximately $33.88{\pm}0.79%$ at 5 mg/mL. Additionally, the B16F10 cellular tyrosinase and melanin synthesis inhibition activities by ALP were $54.21{\pm}4.76$ and $56.74{\pm}6.97%$ at $250{\mu}g/mL$, respectively. Similarly, whitening-related protein tyrosinase, tyrosinase-related protein 1 (TRP-1) and TRP-2, and microphthalmia-associated transcription factor (MITF) were reduced by ALP treatment. These results indicated that ALP could be used as a functional cosmetic ingredient after confirming its whitening activity related to melanin content.

Secretory Expression System of Xylose Reductase (GRE3) for Optimal Production of Xylitol (Xylitol 생산에 최적화된 xylose reductase (GRE3)의 분비발현 시스템)

  • Jung, Hoe-Myung;Kim, Jae-Woon;Kim, Yeon-Hee
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1376-1382
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    • 2016
  • Xylitol is widely used in the food and medical industry. It is produced by the reduction of xylose (lignocellulosic biomass) in the Saccharomyces cerevisiae strain, which is considered genetically safe. In this study, the expression system of the GRE3 (YHR104W) gene that encodes xylose reductase was constructed to efficiently produce xylitol in the S. cerevisiae strain, and the secretory production of xylose reductase was investigated. To select a suitable promoter for the expression of the GRE3 gene, pGMF-GRE3 and pAMF-GRE3 plasmid with GAL10 promoter and ADH1 promoter, respectively, were constructed. The mating factor ${\alpha}$ ($MF{\alpha}$) signal sequence was also connected to each promoter for secretory production. Each plasmid was transformed into S. cerevisiae $SEY2102{\Delta}trp1$, and $SEY2102{\Delta}trp1$/pGMF- GRE3 and $SEY2102{\Delta}trp1$/pAMF-GRE3 transformants were selected. In the $SEY2102{\Delta}trp1$/pGMF-GRE3 strain, the total activity of xylose reductase reached 0.34 unit/mg-protein when NADPH was used as a cofactor; this activity was 1.5 fold higher than that in $SEY2102{\Delta}trp1$/pAMF-GRE3 with ADH1 as the promoter. The secretion efficiency was 91% in both strains, indicating that most of the recombinant xylose reductase was efficiently secreted in the extracellular fraction. In a baffled flask culture of the $SEY2102{\Delta}trp1$/pGMF-GRE3 strain, 12.1 g/l of xylitol was produced from 20 g/l of xylose, and ~83% of the consumed xylose was reduced to xylitol.

Isolation and Identification of a Streptomyces sp. that Produces Antibiotics Against Multidrug - Resistant Acinetobacter baumannii (다제내성 Acinetobacter baumannii의 생장을 억제하는 항생물질을 생산하는 방선균의 분리.동정 및 항균효과)

  • Rhee, Ki-Hyeong
    • Microbiology and Biotechnology Letters
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    • v.39 no.1
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    • pp.37-42
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    • 2011
  • I isolated the actinomycete strain KH223 from soil samples collected from the Kye Ryong mountain area. This strain is antagonistic to the multidrug-resistant Acinetobacter baumannii. KH223 was confirmed as belonging to the genus Streptomyces based on the scanning electronmicroscopy(SEM) observations of the diaminopimelicacid(DAP) type and morphological and physiological characteristics. Comparison of the 16S rDNA nucleotide sequences revealed that KH223 has a relationship with Streptomyces galbus. Production of antibiotics by KH223 was most favorable when cultured on a glucose, polypeptone, and yeast extract(PY) medium for 6 days at 27$^{\circ}C$. The supernatant was found to exhibit an antimicrobial effect on various kinds of bacteria and fungi. Particularly, butanol and ethylacetate extracts of KH223 and cyclo(trp-trp) exhibited significant activity against A. baumannii at concentration ranges of 0.8-12.5 ${\mu}g$/mL, 5.0-25 ${\mu}g$/mL and 12.5${\rightarrow}$100 ${\mu}g$/mL, respectively. Moreover, in contrast to cyclo(trp-trp) had shown to activity against Micrococcus luteus JCM 1464 at the concentration of 12.5 ${\mu}g$/mL, the butanol extract of KH223 showed significant activity against Bacillus subtilis IAM 1069 and Micrococcus luteus JCM 1464 at the concentration of 0.4 and 0.8 ${\mu}g$/mL, respectively. These results suggest that KH223 may have a great potential in the production of new antibiotics to combat multidrug-resistant pathogens and further studies may be warranted for the same.