Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.10.3.1 (tyrosinase)
9,065 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Microporous membranes were used as a support material for enzyme immobilization by a supported aqueous-phase. To test the concept, a model reaction was chosen involving the oxidation of p-cresol by tyrosinase. Tyrosinase was first immobilized in a thin film of water formed on the inner surface of the membrane and then allowed to catalyze p-cresol oxidation in chloroform. By choosing optimal operating conditions, tyrosinase functioned catalytically for more than 6 hours with a stable reaction rate. The reaction rate was highly dependent on water content (water wt./enzyme wt. ratio) and permeation flux. Also, enzyme loading was an important factor for maintaining stable activity. This type of high-rate reactor utilized convective flow through an enzyme immobilized microporous membrane and provided high productivity by reducing mass transport limitations.
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PMID:High-rate membrane supported aqueous-phase enzymatic conversion in organic solvent. 776 81

The inactivation of the catecholase activity of mushroom tyrosinase was investigated under nonaqueous conditions. The enzyme was immobilized on glass beads, and assays were conducted in chloroform, toluene, amyl acetate, isopropyl ether, and butanol. The reaction components were pre-equilibrated for 2 weeks with a saturated salt solution at a water activity of 0.90. The initial reaction velocity varied between 1.3 x 10(3) mol product/((mol enzyme)(min)) in toluene and 8.7 x 10(3) mol product/((mol enzyme)(min)) in amyl acetate. The turnover number varied between 8.1 x 10(3) mol product/mol enzyme in toluene and 7.2 x 10(4) mol product/mol enzyme in amyl acetate. In each solvent, the tyrosinase reaction inactivation parameters were represented by a probabilistic model. Changes in the probability of inactivation were followed throughout the course of the reaction using a second model which relates the reaction velocity to the amount of product formed. These models reveal that the inactivation rate of tyrosinase decreases as the reaction progresses, and that the inactivation kinetics are independent of the quinone concentration in toluene, chloroform, butanol, and amyl acetate. Significant effects of quinone concentration were, however, observed in isopropyl ether. The likelihood of inactivation of the enzyme was found to be greatest toward the beginning of the reaction. In the latter phase of the reaction, inactivation probability was less and tended to remain constant until the completion of the reaction.
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PMID:Tyrosinase inactivation in organic solvents. 1048 31

For the molecular detection of rare tumour cells in clinical samples, real-time reverse transcription-polymerase chain reaction (RT-PCR) offers two important advantages over conventional RT-PCR assays: the results are quantitative and, perhaps more importantly, it facilitates exact sensitivity controls on a per sample basis as well as exact comparison of different assay protocols. We report here on quantitative results obtained with different protocols for RNA isolation and cDNA synthesis for amplification of beta2-microglobulin transcripts using the light cycler system. Furthermore, housekeeping gene-specific PCRs were compared with PCRs specific for an artificial transcript (internal standard) detected simultaneously at a level comparable to the wild-type sequence. Artificial tyrosinase transcripts derived from a vector construct stably transfected into a human lymphoma cell line were used as a model to test the usefulness of artificial internal standards as an alternative to housekeeping genes. The highest RNA yields were obtained using a combination of phenol-chloroform extraction and the High Pure RNA Isolation Kit. Analysing beta2-microglobulin transcript-specific RT-PCRs, the highest sensitivity was obtained for cDNAs generated with Omniscript reverse transcriptase and oligo-p(dT)15 primer. Regarding patient blood samples, RT-PCRs specific for beta2-microglobulin, porphobilinogen deaminase and artificial tyrosinase transcripts provided quantitative data for all, for 18 out of 21, and for 10 out of 21 samples, respectively. Quantification of beta2-microglobulin transcripts by the light cycler system defined the protocol revealing the highest cDNA quality. Comparisons of quantitative data from RT-PCRs specific for beta2-microglobulin, porphobilinogen deaminase and artificial tyrosinase transcripts enabled us to determine a close range for crossing points within which sufficient cDNA quality can be guaranteed, even for the detection of rare transcripts. PCRs specific for the artificial internal standard are ideally suited for cDNA quality assessment on a per sample basis.
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PMID:Reliability of PCR-based detection of occult tumour cells: lessons from real-time RT-PCR. 1147 25

The inhibitor of tyrosinase activity in black rice bran was investigated. The methanol extract from black rice bran was re-extracted with hexane, chloroform, ethyl acetate, or water. The ethyl acetate extract had the most potent inhibition against tyrosinase activity by 80.5% at a concentration of 0.4 mg/mL. Inhibitory compound in the ethyl acetate fraction was isolated by silica gel column chromatography, and identified as protocatechuic acid methyl ester (compound 1) by GC, GC-MS, IR, and 1H and 13C NMR spectroscopy. Compound 1 inhibited 75.4% of tyrosinase activity at a concentration of 0.50 micromol/mL. ID(50) (50% inhibition dose) value of compound 1 was 0.28 micromol/mL. To study the structure-activity relationship, protocatechuic acid (2), vanillic acid (3), vanillic acid methyl ester (4), isovanillic acid (5), isovanillic acid methyl ester (6), veratric acid (7), and veratric acid methyl ester (8) were also assayed.
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PMID:Tyrosinase inhibitor from black rice bran. 1461 Nov 53

Sensitive amperometric biosensors for phenols compounds, based on tyrosinase (polyphenoloxidase, PPO) immobilized on a Pt electrode in an electropolymerized poly-amphiphilic pyrrole matrix or cross-linked with glutaraldehyde, were constructed and compared. Steady-state amperometric measurements, performed at -50 mV vs. SCE in aqueous phosphate buffer containing LiClO(4) 0.1 M (pH 7) as well as in a chloroform solution containing 0.1 M C(6)H(5)CH(2)N(CH(3))(3)Cl, were used in order to compare the electroanalytical and kinetic parameters of the investigated amperometric biosensors in aqueous and nonaqueous media. It was established that the polypyrrole matrix has a higher efficiency for enzyme retention resulting in higher bioelectrode sensitivity, both in aqueous buffer (690 microA M(-1)) and in chloroform (149 microA M(-1)).
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PMID:Phenols monitoring and Hill coefficient evaluation using tyrosinase-based amperometric biosensors. 1521 46

Long-chain esters 1 and 2 have been isolated from the chloroform-soluble fraction of Amberboa ramosa and their structures assigned to be methyl 2beta(2S)-hydroxyl-7(E)-tritriacontenoate (1) and methyl 2beta(2S)-O-beta-D-galactopyranosyl-7(E)-tetratriacontenoate (2). In addition, tricontane (3) and apigenin (4) are also reported for the first time from this species. The structures were assigned on the basis of 1D and 2D NMR techniques. Compounds 1 and 2 showed strong to moderate inhibitory activity against tyrosinase.
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PMID:Tyrosinase-inhibitory long-chain esters from Amberboa ramosa. 1563 36

Aloe chromone is a group of anti-inflammatory and anti-tyrosinase constituents found in aloe vera leaves. High-speed countercurrent chromatography (HSCCC) is reported for the preparative isolation and purification of a chromone from aloe vera. The crude extract was obtained by a series of pretreatment of aloe vera leaves and extracted from decolorizing active carbon with methanol. Then the extract was distributed between dichloromethane and water, and the organic part was then subjected to HSCCC for the isolation of chromone constituents. The chromone compounds with a high performance liquid chromatographic grade (>95%) was isolated through two step HSCCC separations by employing two solvent systems composed of chloroform-methanol-water and dichloromethane-methanol-water at volume ratios of 4/3/2 and 5/4/2, respectively. The chromone was finally identified as cinnamoyl-C-glycoside chromone by ultraviolet (UV), fast atom bombardment mass spectrometry (FAB-MS), nuclear magnetic resonance (1H NMR and 13C NMR).
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PMID:[Preparative isolation and purification of cinnamoyl-C-glycoside chromone from aloe vera by high-speed countercurrent chromatography]. 1588 78

Several triazine pesticides, such as atrazine, are much more soluble in several organic solvents, such as chloroform, than in water. Our recent research was aimed at analyzing this class of pesticides using tyrosinase OPEE (organic phase enzyme electrodes), exploiting their inhibiting action on the tyrosinase enzyme when operating in water-saturated chloroform medium. In this work we studied the response of a tyrosinase inhibition enzyme sensor to several triazinic (simazine, propazine, terbuthylazine) and benzotriazinic (azinphos-ethyl and azinphos-methyl) pesticides (LOD=0.5x10(-9) mol l(-1)). Recovery trials were also performed in vegetal matrixes (corn, barley, lentils). Lastly, the effect of the solvent (chloroform or water) on the inhibition process was investigated via Hill's equation and the diffusion of analyte from the solvent to the enzyme membrane.
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PMID:Tyrosinase inhibition organic phase biosensor for triazinic and benzotriazinic pesticide analysis (part two). 1632 40

Recent research performed in our laboratory (using a butyrylcholinesterase+choline oxidase enzyme electrode) suggested the validity of the biosensor approach using enzyme inhibition OPEEs (i.e. enzyme electrodes working in organic phase) in the case of organophosphorus and carbamate pesticides, which are poorly soluble in aqueous solutions. Since these pesticides are generally much more soluble in chloroform than in water, the present research aimed at analysing this class of pesticides using a tyrosinase inhibition OPEE operating in water-saturated chloroform medium. The tyrosinase biosensor was assembled using an oxygen amperometric transducer coupled to the tyrosinase enzyme, immobilized in kappa-carrageenan gel. Lastly a detailed comparison between the inhibition monoenzymatic tyrosinase and inhibition bienzymatic (butyrylcholinesterase+choline oxidase) OPEEs was performed and discussed in this work.
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PMID:Organophosphorus and carbamate pesticide analysis using an inhibition tyrosinase organic phase enzyme sensor; comparison by butyrylcholinesterase+choline oxidase opee and application to natural waters. 1738 49

In the present study, we analyzed the antioxidant, antiplatelet aggregation, anti-inflammatory, and tyrosinase inhibitory activities of a variety of solvent extracts of Elaeagnus multiflora Thunb. (Family Elaeagnaceae). Among the solvent extracts of E. multiflora, the ethyl acetate extract (EE) exhibited the highest 1,1-diphenyl-2-picrylhydrazyl radical and xanthine oxidase inhibition activity, as well as the greatest tyrosinase inhibition activity. Only the chloroform extract (CE) inhibited platelet aggregation, and that was a weak effect with 19.29% inhibition at 250 microg/mL, as compared to controls. The CE was also the most potent inhibitor of nitric oxide production among the tested fractions, with almost 100% inhibition at 500 microg/mL. We also detected 2,4-bis(1,1-dimethylethyl)phenol in the CE and EE, via a gas chromatography-mass spectrometry assay. In conclusion, we found that E. multiflora Thunb. has antioxidant and antiplatelet aggregation effects to some extent, and its CE and EE possess potent inhibitory effects on nitric oxide production and tyrosinase activity.
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PMID:Antioxidant activity, anti-inflammatory activity, and whitening effects of extracts of Elaeagnus multiflora Thunb. 1747 76


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