• Title/Summary/Keyword: DOPA solution

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Drinking Wateres Effects on Tyrosinase Activities (물리 Tyrosinase의 활성도에 미치는 영향)

  • Rhee, Byung-Chul;Lee, Zong-Liong;Lee, Duk-Soo;Kim, Yil
    • Journal of Environmental Health Sciences
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    • v.22 no.1
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    • pp.1-4
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    • 1996
  • I have conducted two testings to find out which water is better for drinking water. First, I made 20 mM L-DOPA solutions by solving L-DOPA (3,4-Dihydroxyphenylalanine) in tap water, Waters' mineral water and reverse osmotic water. Then I measured activities after adding Tyrosinase (purifide enzyme, step 3), which was extracted from Salanum melongena(mad apple), in each L-DOPA solution. Second, I solved 0.1, 0.5 and 0.9% salt in each 20 mM L-DOPA distilled water to measure activity of each salt solution. The results of the testings are as follows: 1. 10 minutes after adding Salanum melongena(mad apple) tyrosinase in each L-DOPA solution, activity of Waters' mineral water was 0.867 tap water 0.777 and reverse osmotic water 0.742. 2. Activity of Waters' mineral water was higher than that of tap water by 10.4% and higher then reverse osmotic by 14.4%. 3. Activity of Waters' mineral water was much higher than that of 0.9% salt water by 41.8%. 4. The optimum pH of Salanum melongena (mad apple) tyrosinase is 9.0. Most enzymes working in the human metabolism are alkaline and body fluids' pH also alkaline. In conclusion, an alkaline water is believed better than an acidic water for drinking.

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Selective 3,4-Dihydroxyphenylalanine Analysis in Human Urine as Ethoxycarbonyltert-butyldimethylsilyl Derivatives by Gas Chromatography-Mass Spectrometry

  • Paik, Man-Jeong;Nguyen, Duc-Toan;Yoon, Jae-Hwan;Cho, In-Seon;Shim, Woo-Young;Kim, Kyoung-Rae;Cho, Ki-Hong;Choi, Sang-Dun;Lee, Gwang
    • Bulletin of the Korean Chemical Society
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    • v.32 no.3
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    • pp.977-980
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    • 2011
  • A new analytical method for measurement of 3,4-dihydroxyphenylalanine (DOPA) in human urine was developed. DOPA from an aqueous solution was converted into an ethoxycarbonyl (EOC) derivative. A tertbutyldimethylsilyl (TBDMS) reaction under anhydrous conditions was then attempted for analysis by gas chromatography-mass spectrometry in selected ion monitoring mode. A new mass spectral data on DOPA as a tri-EOC/mono-TBDMS derivative was built. This method showed good linearity (r ${\geq}$ 0.999), precision (% relative standard deviation = 3.1-9.2), and accuracy (% relative error = -7.2-8.8), with a detection limit of 0.05 ng/mL. This selective and accurate method of DOPA analysis will be useful for biochemical monitoring of various neurological disorders including Parkinson's disease in biological fluids.

Production of L-DOPA by Thermostable Tyrosine Phenol-lyase of a Thermophilic Symbiobacterium Species Overexpressed in Recombinant Escherichia coli

  • Lee, Seung-Goo;Ro, Hyeon-Su;Hong, Seung-Pyo;Kim, Eun-Hwa;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.6 no.2
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    • pp.98-102
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    • 1996
  • A thermostable tyrosine phenol-lyase gene of a thermophilic Symbiobacterium species was cloned and overexpressed in Escherichia coli in order to produce the biocatalyst for the synthesis of 3, 4-dihy-droxyphenyl-L-alanine (L-DOPA). The substrates used for the synthetic reaction were pyrocatechol, so-dium pyruvate, and ammonium chloride. The enzyme was stable up to $60^{\circ}C$, and the optimal temperature for the synthesis of L-DOPA was $37^{\circ}C$ . The optimal pH of the reaction was about 8.3. Enzyme activity was highly dependent on the amount of ammonium chloride and the optimal concentration was estimated to be 0.6 M. In the case of pyrocatechol, an inactivation of enzyme activity was observed at con-centrations higher than 0.1 M. Enzyme activity was increased by the presence of ethanol. Under op-timized conditions, L-DOPA production was carried out adding pyrocatechol and sodium pyruvate to the reaction solution intermittently to avoid substrate depletion during the reaction. The concentration of L-DOPA reached 29.8 g/l after 6 h, but the concentration didn t increase further because of the formation of byproducts by a non-enzymatic reaction between L-DOPA and pyruvate.

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Determination of L-Dopa by Spectrofluorimetry Using Co-fluorescence Enhancer (보조 형광증가제를 이용한 L-Dopa의 형광분광법적 정량)

  • Lee, Sang Hak;Ahn, Jung Mi
    • Journal of the Korean Chemical Society
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    • v.44 no.6
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    • pp.541-546
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    • 2000
  • Methods to determine L-dopa(L-3,4-dihydroxyphenylalanine) in aqueous solution by spectrofluorimetry based upon the ligand sensitized luminescence of Tb(III) ion L-dopa complex have been studied. Tb(III) ion and Lu(III) ion were used as ligand sensitized fluorescencer and co-fluorescence enhancer, respectively. The effects of excitation wavelength, pH, concentration of Tb(III) ion, concentration of Lu(III) ion and emission wavelength on the fluorescence intensity were investigated. The fluorescence intensity of the Tb(III) ion L-dopa complex was further increased with addition of Lu(III) ion. The calibration curve for L-dopa was linear over the range from 5.0 ${\times}$ $10^{-7}$ M to 1.0${\times}$ $10^{-4}$ M and the detection limit was 4.0 ${\times}$ $10^{-8}$ M under the optimal experimental conditions of 300 nm, 8.0, 1.0 ${\times}$ $10^{-4}$ M and 545 nm for excitation wavelength, pH, concentration of Tb(III) ion and emission wavelength, respectively. When Lu(III) ion was added to Tb(Ⅲ) ion L-dopa complex, the concentration range of linear response and detection limit were 1.0${\times}$$10^{-8}$ M to 2.0 ${\times}$ $10^{-4}$ M and 1.0 ${\times}$ $10^{-9}$ M, respectively under the optimal experimental conditions of 300 nm, 8.5, 1.0 ${\times}$ $10^{-5}$ M, 1.0 ${\times}$ $10^{-5}$ M, 545 nm for excitation wavelength, pH, concentration of Tb(III) ion, concentration of Lu(III) ion and emission wavelength, respectively.

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Production of 3,4-dihydroxyphenyl-L-alanine by Using the ${\beta}$-Tyrosinase of Citrobacter freundii Overexpressed in Recombinant Escherichia coli. (재조합 대장균에서 과발현된 Citorbacter Freundii KCTC2006 유래의 ${\beta}$-Tyrosinase를 이용한 3,4-Dihydroxyphenyl-L-alannine의 생산)

  • Lee, Seung-Goo;Ro, Hyeon-Su;Hong, Seung-Pyo;Lee, Kyu-Jong;Wang, Ji-Won;Tae, Dong-Nyeon;Uhm, Ki-Nam;Bang, Sang-Gu;Kim, Young-Jun;Sung, Moon-Hee
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.44-49
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    • 1996
  • By using the ${\beta}$-tyrosinase of Citrobacter freundii KCT2006, which was cloned and overexpressed in Escherichia coli, 3,4-dihydroxy phenyl-L-alanine (L-DOPA) was synthesized efficiently from pyrocatechol, sodium pyruvate, and ammonium acetate. Optimal temperature and pH for the reaction were determined to be about 18$^{\circ}C$ and 8.5, respectively. The effects of substrate concentrations were also examined at different concentrations of ammonium acetate, sodium pyruvate, and pyrocatechol. Ammoniumacetate and sodium pyruvate increased the reaction rate until the concentrations reached to 300mM and 50mM, respectively. Although pyrocatechol showed the optimal concentration at 20mM, it was controlled between 20mM and 50mM to avoid the depletion of substrate during the enzymatic synthesis. Meanwhile the synthetic rate was improved about 20% when ethanol was included in the reaction solution. Based on above results, a reaction medium for the productin of L-DOPA was prepared and incubated with 1 unit/ml of ${\beta}$-tyrosinase. Pyrocatechol and sodium pyruvate was added to the reaction solutin intermittently to avoid the substrate depletion during the enzymatic reaction. After 24 hour of reaction, 31.6g/l of L-DOPA was accumulated in the reaction solution as soluble and precipitated ones and the conversion yield was about 85.2%.

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Rec-assay and DNA-breaking Action on the Engymatic Browning Reaction Products -Influence of Cupric Ion Concentration- (효소적(酵素的) 갈변반응(褐變反應) 생성물(生咸物)에 대(對)한 Rec-assay 및 DNA 절단(切斷) 작용(作用) -동(銅) Ion 농도(濃度)의 영향(影響)-)

  • Ham, S.S.;Park, B.K.;Lee, S.Y.;Lee, J.H.;Kang, C.K.;Lee, D.S.;Omura, Hirohisa
    • Applied Biological Chemistry
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    • v.27 no.4
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    • pp.264-270
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    • 1984
  • In order to obtain mutagenic data of enzymatic browning reaction products, we investigated their mutagenicity. The rec-assay with Bacillus subtilis strains $H17(rec^+)$ and $M45(rec^-)$ were carried out with their spores. Detection of double strand breakage in calf-thymus DNA was investigated into sample solution with and without $Cu^{2+}$ by the agarose slab gel electrophoresis. In the rec-assay, catechol, pyrocatechol, DL-dopa, 3,4-dihydroxytoluene, hydroquinone, hydroxyhydroquinone with and without $Cu^{2+}$ in 0.05M ana 0.1M at the enzymatic browning reaction products stowed mutagenic action. And also browning solution of 0.05M hydroxyhydroquinone and catechol with $Cu^{2+}$, hydroquinone with and without $Cu^{2+}$ of 0.1M at the enzymatic browning reaction products were strong mutagenic action. The DNA breakability of the enzymatic browning reaction products of 0.1M pyrogallol was stronger than that of 0.05M pyrogallol browning solution with $Cu^{2+}$ and 3,4-dihydroxytoluene browning solution was stronger than that of 0.01M 3,4-dihydroxytoluene browning solution.

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Recent Progress in Mussel-inspired Catechol-conjugated Chitosan Hemostats (홍합 모사 카테콜기가 도입된 키토산 지혈제 연구 동향)

  • Seongyeon Jo;Soomi Kim;Chanwoo Park;Seungwon Hong;Hong Kee Kim;Ji Hyun Ryu
    • Journal of Adhesion and Interface
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    • v.24 no.4
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    • pp.113-119
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    • 2023
  • Since it was reported that the unusual amino acid DOPA in synergy with lysine and histidine residues found in mussel adhesive proteins plays a pivotal role in mussel adhesion in underwater environments, there has been a burgeoning development of various catecholamines-based adhesives for biomedical applications. Among these, catechol-conjugated chitosan, containing catecholamine, featuring multiple catechol groups within its aminerich chitosan backbone, has found versatile utility in fields, such as tissue adhesion, wound dressing, tissue healing, hemostats, drug delivery systems, and tissue engineering scaffolds. Significantly, chitosan-catechol is a mussel-inspired material approved by both US Food and Drug Administration (FDA) and KR Ministry of Food and Drug Safety (MFDS) for its effectiveness in hemostasis. This review focuses on 1) general aspects of catechol-conjugated chitosan, highlighting catechol group integration into chitosan backbones, 2) examination of proposed mechanisms of hemostasis, and 3) exploration of diverse physical forms, including solution, hydrogels, patches, and thin films with practical applications inapplicable to hemostasis.

The Synthetic Melanin Nanoparticles Having An Excellent Binding Capacity of Heavy Metal Ions

  • Kim, Da Jeong;Ju, Kuk-Youn;Lee, Jin-Kyu
    • Bulletin of the Korean Chemical Society
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    • v.33 no.11
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    • pp.3788-3792
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    • 2012
  • Spherical-shape melanin nanoparticles with good water-dispersibility were successfully synthesized by a simple oxidation polymerization of 3,4-dihydroxy-phenylalanin (DOPA) with $KMnO_4$. Similar features to those known from natural and synthetic melanin polymers were observed from prepared melanin nanoparticles by FT-IR, UV-Vis., and ESR spectroscopic methods. Their binding ability with several heavy metal ions from aqueous solution was quantitatively investigated, and the maximum binding capacities with melanin nanoparticles to lead, copper, and cadmium ions were obtained as 2.45, 2.17 and 1.88 mmol/g, respectively, which are much larger values than those reported from natural and synthetic melanin polymers. The large binding capacity and fast binding rate of melanin nanoparticles to metal ions can make them an excellent candidate for the remediation of contaminated water.

Functional Cosmetic Characteristics of Momordica charantia Fruit Extract (여주 열매 추출물의 기능성화장품 소재 특성)

  • Kim, Hyun-Woo;Shin, Hyejin;Hwang, Danbi;Lee, Jieun;Jeong, Hyangli;Kim, Donguk
    • Korean Chemical Engineering Research
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    • v.53 no.3
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    • pp.289-294
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    • 2015
  • In this research, water and ethanol extract of Momordica charantia shells, fruits and seeds were tested to see possibility as natural functional cosmetic agent. Water and ethanol extract showed 69.45 mg/g and 70.87 mg/g polyphenol concentration, respectively. Momordica charantia water and ethanol extracts did not indicate cell toxicity up to $1,000{\mu}g/ml$ concentration in MTT assay. Tyrosinase inhibition effects of water and ethanol extract were lower than arbutin, however, ethanol extract showed better DOPA oxidation inhibition effect than arbutin. Elastase inhibition effects of ethanol extract displayed similar efficacy with adenosine at higher concentrations. Solution formulations (5% extract) were stable for 28 days in both extracts, however, lotion formulation (1% extract) showed considerable variation in viscosity whereas ethanol extraction indicated relative stability. In conclusion, water and ethanol extract of Momordica charantia shells, fruits and seeds indicated strong possibility for whitening and antiwrinkle functional cosmetic ingredient.

Experimental studies on the Whitening and Anti-allergic effect of various Herbal medicines that clear heat (수종의 청열약물의 미백 및 항알러지 효과에 대한 실험적 연구)

  • Lee, Myoung-Gyu;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.20 no.3
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    • pp.71-81
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    • 2007
  • Objective : In this study, herbal medicine(GJE, Gardenia jasminoides Ellis; HCT, Houttuynia cordata Thunb.; CIL, Chrysanthemum indicum Linne; PMS,Paeonia moutan Sims, P. subfruticosa Makino; APL, Agrimonia pilosa Ledebour) were screened for their inhibitory activities against Tyrosinase and PMA plus A23187-induced $TNF-{\alpha}$, IL-6, IL-8 productions in HMC-1 cells to reveal their skin -whitening and anti-allergic effect. Method : To investigate Tyrosinase inhibition we treated Mushroom Tyrosinase(Fluka, 93898) $10{\mu}{\ell}$ and 7.5mM Tyrosine (Sigma, T3754) $20{\mu}{\ell}$ with 80% ethanol medicine extracts. Then we observed 96well micro plate extinction at 490nm. In the next experiment, to investigate Anti-allergic effect we blended cultured Human Mast Cells(HMC-1) with medicine extracts. We treated the blended solution with Phorbol 12-myristate 13-acetate(PMA) and A23187, then observed $TNF-{\alpha}$, IL-6, IL-8 by ELISA (enzyme-linked immunosorbent assay) at 450nm. Results : In inhibiting Tyrosinase the results are as follows. 1. We observed 22% inhibition of Mushroom Tyrosinase at $500{\mu}g/m{\ell}$ concentration of GJE extracts. 2. We also could observe that the decreased Mushroom Tyrosinase activities in HCT, CIL extracts. In inhibiing $TNF-{\alpha}$, IL-6, IL-8 productions in HMC-1 cells the results are as follows. 1. Of the extracts examined, HCT, PMS, APL extracts showed over 50% inhibitions of Cytokines at $200{\mu}g/m{\ell}$ concentration. 2. In particular, APL extracts showed the best inhibitory effect on Cytokine productions in a dose-dependent manner. Conclusion : These results suggest that GJE extracts contributes to the anti melanin activities and represent a potential source of whitening agent. Thus these herbal medicines suggest novel drugs on anti-allergic effects.

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