• Title/Summary/Keyword: TRP1

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Cloning and Sequence Analysis of the trpB, trpA and 3' trpC(F) Gens of Vibrio metschnikovii Strain RH530 (Vibrio metschnikovii 균주 RH530의 trpB, trpA 그리고 3' trpC(F) 유전자의 클로닝 및 염기서열 결정)

  • Kwon, Yong-Tae;Kim, Jin-Oh;Yoo, Young-Dong;Rho, Hyune-Mo
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.120-125
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    • 1994
  • The genes, trpB, trpA and 3’ trpC(F) of Vibrio metschnikovii strain RH530 were cloned and sequenced. The trpB and trpA genes had open reading frames of 1,173 bp and 804 bp encoding 391 and 268 amino acids, respectively. The trpB and trpA genes had conventional ribosome-binding sequences and overlapped with each other by one nucleotide, suggesting that these two genes are translationally coupled. 115 nucleotide upstream the trpB start codon, tjere was an incomplete open reading frame of the 3’-end of the trpC(F). The amino acid sequences of trpB, trpA and trpC(F) of V. metschnikovii RH530 had identities of 64.2%, 82.4% and 73.7% respectively, for those of V. parahaemolyticus; 58.7%, 72.3% and 54.9%, respectively, for Salmonella typhimurium; and 42.6%. 54.1% and 12.5%, respectively, for brevibacterium lactofermentum. The genetic organization of these genes, especially in the noncoding region between trpC(F) and trpB, was distinct from that of Enterobacteriaceae.

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Isolation of the Phosphoribosyl Anthranilate Isomerase Gene (TRP1) from Starch-Utilizing Yeast Saccharomycopsis fibuligera

  • Park, Eun-Hee;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1324-1327
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    • 2015
  • The nucleotide sequence of the TRP1 gene encoding phosphoribosyl anthranilate isomerase in yeast Saccharomycopsis fibuligera was determined by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed the presence of an uninterrupted open-reading frame of 759 bp, including the stop codon, encoding a 252 amino acid residue. The deduced amino acid sequence of Trp1 in S. fibuligera was 43.5% homologous to that of Komagataella pastoris. The cloned TRP1 gene (SfTRP1) complemented the trp1 mutation in Saccharomyces cerevisiae, suggesting that it encodes a functional TRP1 in S. fibuligera. A new auxotrophic marker to engineer starch-degrading yeast S. fibuligera is now available. The GenBank Accession No. for SfTRP1 is KR078268.

Antioxidant Effect of Nelumbo nucifera G. Leaf Extract and Inhibition of MITF, TRP-1, TRP-2, and Tyrosinase Expression in a B16F10 Melanoma Cell Line (연잎 추출물의 항산화 활성 및 멜라노마 세포(B16F10)에서 MITF, TRP-1, TRP-2, tyrosinase의 발현 저해 효과)

  • Yoo, Dan-Hee;Joo, Da-Hye;Lee, Soo-Yeon;Lee, Jin-Young
    • Journal of Life Science
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    • v.25 no.10
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    • pp.1115-1123
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    • 2015
  • The purpose of this study was to investigate the potential of Nelumbo nucifera G. leaf (NNL) extract as a cosmetic additive. The electron-donating ability of the NNL extract at a concentration of 1,000 μg/ml was 67.83%. In xanthine oxidase, the inhibition effect of the NNL extract was 92.7% at the same concentration. For whitening effects, tyrosinase inhibition effect of NNL extract was 42.7% at a 1,000 μg/ml concentration. The cell toxicity of the NNL extract was examined in melanoma cells (B16F10) using a 3-[4, 5–dimethyl–thiazol–2–yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay. The cell toxicity assay revealed that the NNL extract had a toxicity of 81.61% at a concentration of 1,000 μg/ml The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), and tyrosinase protein expression inhibitory effect by Western blot of NNL extract were measured by a Western blot at concentrations of 25, 50, and 100 μg/ml. At a 100 μg/ml concentration of the NNL extract, the expression of the MITF, TRP-1, TRP-2, and tyrosinase protein was decreased by 69.59%, 27.74%, 67.33%, and 67.78% respectively. The MITF, TRP-1, TRP-2 and tyrosinase mRNA expression inhibitory effect were measured by reverse transcription- polymerase chain reaction (PCR) at concentrations of 25, 50, and 100 μg/ml. GAPDH was used as a positive control. At a concentration of 100 μg/ml of the NNL extract, the expression of MITF, TRP-1, TRP-2, and tyrosinase mRNA was decreased by 67.51%, 71.36%, 85.74%, and 83.64%, respectively. These findings suggest that the NNL extract has antioxidant and whitening effects and that it has great potential as a cosmetic ingredient.

Inhibitory Effect of Korean Fermented Soybean (Chungkookjang) Extract and Genistein Against Trp-P-1 Induced Genotoxicity in HepG2 Cells

  • Song, Eun Jeong;Kim, Nam Yee;Heo, Moon Young
    • Journal of Food Hygiene and Safety
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    • v.32 no.3
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    • pp.171-178
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    • 2017
  • This study evaluated the protective effect of Chungkookjang (CKJ) extract, a Korean traditional fermented soybean product made from Bacillus species in rice straw and boiled soybean, and one of its main flavonoids, genistein, against Trp-P-1 induced cytotoxicity and DNA damage in HepG2 cells. CKJ and genistein exhibited protective effect against Trp-P-1 induced cytotoxicity and Trp-P-1 induced DNA single strand breaks. CKJ and genistein inhibited Trp-P-1 induced CYP1A1 and CYP1A2 transcription in HepG2 cells. Our results indicated that CKJ and genistein have the protective effect against Trp-P-1 induced cytotoxicity and DNA damage. Via inhibiting expression of CYP1A1 and CYP1A2. CKJ can be used as a promising functional food material that prevents the genotoxicity induced by carcinogens produced by the heat treatment of foods such as heterocyclic amines (HCAs) that cause genomic instability.

Anti-oxidant Function and Inhibitory Effects of the Expression of MITF, TRP-1, TRP-2 and Tyrosinase of Sesamum indicum L. in B16F10 Melanoma Cells (참깨 추출물의 항산화 활성 및 melanoma cell (B16F10)에서 MITF, TRP-1, TRP-2, tyrosinase 의 발현 저해)

  • Yoo, Dan-Hee;Joo, Da-Hye;Lee, Jin-Young
    • Journal of Life Science
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    • v.27 no.3
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    • pp.318-324
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    • 2017
  • This study was performed to improve the antioxidant and skin-whitening activities of 70% ethanol extract from Sesamum indicum L. (SIL). The electron-donating ability of the SIL extract was 71.7% at a concentration of $1,000{\mu}g/ml$. The whitening effects that was measured by tyrosinase inhibition assay. As a result, SIL extract was shown 42% at $1,000{\mu}g/ml$ concentration. The cell toxicity on B16F10 melanoma cells of SIL of 70% ethanol extract showed 84.3% at $1,000{\mu}g/ml$ concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase relate protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2) and Tyrosinase protein and mRNA expression inhibitory effect of SIL extract were measured by western blot and reverse transcription- polymerase chain reaction (PCR) at 50, 250, $500{\mu}g/ml$ concentration. Consequently, the MITF, TRP-1, TRP-2, Tyrosinase protein expression inhibitory effect of SIL extract was decreased by 68.3%, 39.2%, 89.7%, 22.3%, respectively, at $500{\mu}g/ml$ concentration. Moreover, MITF, TRP-1, TRP-2, Tyrosinase mRNA expression inhibitory effect by reverse-transcription-PCR of SIL extract was decreased by 81.8%, 66.5%, 84.2%, 68.1%, respectively, at $500{\mu}g/ml$ concentration. Therefore, we excellently identified the antioxidant activities and whitening effect of SIL extract, and this finding suggested that SIL extract has great potential as a cosmetic ingredients.

Modigication of host cells and Expression of Recombinant E. coli trp plasmids for the increased Production of Tryptophan in Klebsiella pneumoniae (Klebsiella pneumoniae에서 트립토판 생산증대를 위한 숙주개발 및 재조합 trp plasmid의 발현)

  • 지연태;홍광원;박장현;이세영
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.46-51
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    • 1987
  • In order to increase the production of tryptophan by maximizing expression of recombinant trp plasmid, Klebsiella pneumoniae KC 105(pheA tyrA trpE trpR tyrR) was genetically modified. KC 107, inosine monophospate(IMP) auxotroph from KC 105 and KC 108, histidine(His) auxotroph from KC 107 were also derived respectively to increase phosphoribosylpyrophosphate(PRPP) production which is required for tryptophan biosynthesis. From KC 107 phosphoribosylpyrophosphate consumption which is required for tryptophan biosynthesis. From KC 107 and KC 108, KC 109 and KC 110, both arginine auxotrophs were derived respectively. To investigate the expression of recombinant trp plasmid in the selected K. pneumoniae mutants, the auxotrophic mutants were transformed with recombinant trp plasmids pSC 101-$trpE^{FBR}$, pSC 101-trpL(.DELTA.att) $trpE^{FBR}$ (pSC 101-trp-AF). Amount of tryptophan produced and activities of tryptophan synthase of $trpR^{-}$ mutant (KC 100) and $tyrR^{-}$ mutnat(KC 105) containing recombinant plasmid pSC 101-trp operon were increased by 30-40% as compared with KC 99(pheA tyrA trpE) containing recombinant plasmid pSC 101-trp operon. Activities of tryptophan synthase and production of tryptophan of KC 108 ($His^{-}$) and KC 109($Arg^{-}$) containing recombinant plasmid pSC 101-trp operon were increase by two-fold as compared with KC 107 containing pSC 101-trp operon.

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Antioxidant and Whitening Activities of Chlorogenic Acid, Quercetin, and Quercitrin from the Fruit of Vaccinum oldhami (정금나무 열매(Fruit of Vaccinum oldhami)의 분리 정제물(클로로겐산, 퀘르세틴 및 퀘르시트린)에 관한 항산화 및 미백활성 검증)

  • Jung-Woo Chae;Min-Jeong Oh;Hyeon-Ji Yeom;Jin-Young Lee
    • Journal of Life Science
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    • v.33 no.2
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    • pp.115-128
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    • 2023
  • The fruit of Vaccinum oldhami was separated and purified to obtain the compounds chlorogenic acid (CA), quercetin (QT), and quercitrin (QR). The electron-donating abilities of CA, QT, and QR at 1,000 ㎍/ml were 91.9%, 89.9%, and 77.4%, respectively QT and QR showed 99.5% and 91.4% ABTS+ radical scavenging ability at a 1,000 ㎍/ml concentration, respectively, and CA showed a 95% ability or higher at 100 ㎍/ml. Regarding tyrosinase inhibitory activity, CA, QT, and QR exhibited 29.5%, 34.7%, and 23.7% efficacy, respectively, at 1,000 ㎍/ml. Regarding the cell viability for melanoma cells (B16F10) assessed through MTT assay, CA, QT, and QR showed cell a viability of 80% or more at 100 ㎍/ml. To measure the deterrent of protein expression, CA affected TRP-1 and TRP-2 in accordance with increases in concentration. The protein expression inhibition rate of QT was excellent for TRP-1, TRP-2, and tyrosinase. CA was confirmed to have an excellent mRNA expression inhibitory effect against MITF, and the amount of mRNA expression of TRP-1, TRP-2, and tyrosinase decreased with an increase in the CA concentration. As the concentration of QT increased, the mRNA expression of MITF, TRP-2, and tyrosinase decreased. QR decreased the amount of mRNA as the QR concentration increased. The excellent antioxidant and whitening effects of CA, QT, and QR were thus confirmed.

Inhibitory Efficacy of Smilax china L. on MITF, TRP-1, TRP-2, Tyrosinase Protein and mRNA Expression in Melanoma Cell (B16F10) (멜라노마 세포(B16F10)에서 청미래 덩굴 뿌리 추출물의 MITF, TRP-1, TRP-2, tyrosinase 단백질 및 mRNA 발현 억제 효과)

  • Lee, Soo-Yeon;Yoo, Dan-Hee;Joo, Da-Hye;Jo, Hui-Seon;Lee, Jin-Young
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.1-8
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    • 2016
  • The purpose of this study was to assess the whitening effects of an extract from Smilax china L., which is a vine shrub belonging to the lily family. With regard to the whitening effects, 70% ethanol and water extracts from Smilax china L. showed more than 77.6% and 40.2% tyrosinase inhibition at a concentration of $1,000{\mu}l$. Furthermore, the 70% ethanol extract showed cytotoxicity of 89% at a concentration of $100{\mu}g/ml$ in melanoma cells. Western blot showed that the inhibitory effect of the 70% ethanol extract on MITF, TRP-1, TRP-2, and tyrosinase protein expression decreased by 89.9%, 46.2%, 57.6%, and 55.8%, respectively, at a concentration of $50{\mu}g/ml$. Moreover, reverse transcription-PCR showed that the inhibitory effect of the 70% ethanol extract on MITF, TRP-1, TRP-2, and tyrosinase mRNA expression decreased by 78.5%, 58.0%, 78.8%, and 70.8%, respectively, at the same concentration of $50{\mu}g/ml$ concentration. Further, realtime PCR showed that the 70% ethanol extract-induced decrease in MITF, TRP-1, TRP-2, and tyrosinase quantitative mRNA expression rate was concentration-dependent. The findings suggest that the extract from Smilax china L. has great potential as a cosmetic ingredient with whitening effects.

Whitening Effect of Poria cocas Ethanol Extract by Inhibition of Melanin Synthesis (백복령 주정 추출물의 멜라닌합성 억제를 통한 미백효과)

  • Park, Hye-Jung;Kwon, Eun-Jeong;Kim, Moon-Moo;Lee, Kyeong Rok;Hong, Il;Lee, Do Gyeong;Oh, Yunghee
    • Journal of Life Science
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    • v.24 no.5
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    • pp.485-490
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    • 2014
  • Poria cocas has been reported to be effective in skin whitening. However, the direct effect of P. cocas ethanol extracts (PCEE) on melanin synthesis has not been scientifically studied. To elucidate the direct effect of PCEE on melanogenesis, a 3,4-dihydroxyindole-2-carboxylic acid (DOPA) synthesis assay, tyrosinase activity assay, and Western blotting for melanogenic proteins, including tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 were performed in mouse B16F1 cells. The results revealed that PGEE inhibited melanin production in a dose-dependent manner by blocking the synthesis of DOPA. Although the activation of tyrosinase was not affected, the expression levels of TRP-1 and TRP-2 were controlled. These results suggest that PCEE has a whitening effect, indicating that it may be a useful agent in the development of whitening cosmetics.

Antimutagenic Effect of Rresveratrol on Trip P-1 in Salmonella typhimurium TA98 (Trp P-1 변이원성에 대한 Resveratrol의 항돌연변이 효과)

  • 장귀현;안병용;권용주;최동성
    • The Korean Journal of Food And Nutrition
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    • v.14 no.4
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    • pp.329-332
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    • 2001
  • The antimutagenic activity of resveratrol on the mutagenicity induced by Trp P-1 (3-amino-1,4-dime-thyl-5H-pyrido{4,3-b}indole) was studied using the Ames test with Salmonella typhimurium TA98 and 100. Trp P-1 showed strong mutagenecity in S. typhimurium TA98, but was higly decreased mutagenecity in S. typhimurium TA100. This result suggests that the mutagenecity of Trp P-1 can be mainly induced by the DNA lesions causing frame shift. Resveratrol itself did not show antibacterial effect upon 300 $\mu\textrm{g}$/assay. Resveratrol showed the strongest inhibitory effect with dose dependent manner on the mutagenicity induced by Trp P-1. The inhibition rates of resveratrol at concentration of 2, 10, 25, 50, 100, 300 $\mu\textrm{g}$/assay were 13%, 37%, 52%, 65%, 81%, 89%, respectively.

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