Biomedical subjects

Estrella Núñez-Delicado

Publications and source records attributed to Estrella Núñez-Delicado.

4 recordsLinked to original sources

PubMed2006

Determination of stoichiometric coefficients and apparent formation constants for beta-cyclodextrin complexes of trans-resveratrol using reversed-phase liquid chromatography.

The complexation of trans-resveratrol with beta-cyclodextrin (beta-CD) was investigated using reversed-phase liquid chromatography and mobile phases to which beta-CD was added. The decrease in the retention times with increasing concentrations of beta-CD (0-2.5 mM) showed that trans-resveratrol forms a 1:1 complex with beta-CD, while the apparent formation constants (K(F)) were strongly dependent of the water-methanol proportion of the mobile phase employed. A slight decrease in K(F) was observed at 50-55% water in the mobile phase but a large increase from 55 to 70% water. The values of K(F) for the trans-resveratrol-beta-CD interaction decreased when the temperature was raised from 20 to 37 degrees C. In order to gain information about the mechanism aspect of the trans-resveratrol affinity for beta-CD, the thermodynamic parameters of the complexation were obtained. Complex formation of trans-resveratrol with beta-CD (DeltaG degrees =-17.01 kJ/mol) is largely driven by enthalpy (DeltaH degrees = -30.62 kJ/mol) and slightly entropy changes (DeltaS degrees = -45.68 J/mol K).

Chromatography, High Pressure Liquid↗

PubMed2005

Polyphenol oxidase from Dominga table grape.

Dominga grape polyphenol oxidase (PPO) was extracted using phase partitioning with Triton X-114. The enzyme was obtained in latent state and could be optimally activated by the presence of 0.2% sodium dodecyl sulfate (SDS) at pH 6.0. In the absence of SDS, the enzyme showed maximum activity at acid pH. The kinetic parameters of the enzyme at pH 3.0 and 6.0 in the presence of SDS were calculated. The effect of several inhibitors was studied, tropolone being the most effective with a K(i) value of 18 muM. The effect of cyclodextrins was also studied, and the complexation constant K(c) between G(2)-beta-cyclodextrins and 4-tert-butylcatechol was calculated using the enzymatic method (K(c) = 13960 M(-)(1)). The evolution of the color parameters (L, a, b) of liquefied grape berries was inhibited by inhibitors of PPO activity, such as diethyldithiocarbamate, metabisulfite, and G(2)-beta-cyclodextrins, indicating that enzymatic browning by PPO is the main process involved in the browning of Dominga grape juice at room temperature.

Catechol Oxidase↗

PubMed2003

Partial purification of latent persimmon fruit polyphenol oxidase.

Persimmon fruit polyphenol oxidase (PPO) was partially purified using a combination of phase partitioning with Triton X-114 and ammonium sulfate fractionation between 50 and 75%. The enzyme, which showed both monophenolase and diphenolase activities, was partially purified in a latent form and could be optimally activated by the presence of 1 mM sodium dodecyl sulfate (SDS) with an optimum pH of 5.5. In the absence of SDS, the enzyme showed maximum activity at acid pH. SDS-PAGE showed the presence of a single band when L-DOPA was used as substrate. The apparent kinetic parameters of the latent enzyme were determined at pH 5.5, the V(m) value being 15 times higher in the presence of SDS than in its absence, whereas the K(M) was the same in both cases, with a value of 0.68 mM. The effect of several inhibitors was studied, tropolone being the most active with a K(i) value of 0.45 microM. In addition, the effect of cyclodextrins (CDs) was studied, and the complexation constant (K(c)) between 4-tert-butylcatechol (TBC) and CDs was calculated using an enzymatic method. The value obtained for K(c) was 15580 M(-1).

Ammonium Sulfate↗

PubMed2003

Anomalous oxidation of MDL 73,404 by horseradish peroxidase.

3,4-Dihydro-6-hydroxy-N,N,N-2,5,7,8-heptamethyl-2H-1-benzopyran-2-ethanaminium-4-methylbenzene sulfonate (MDL 73,404) is a cardioselective water-soluble quaternary ammonium analogue of Vitamin E which is synthesized to augment the antioxidant defence in situations of free radical injury such as myocardial infarction/reperfusion. Its oxidation by any peroxidative enzyme has not been studied kinetically. This paper describes its enzymatic oxidation by horseradish peroxidase (HRP). The activity was followed spectrophotometrically at 255nm, and the experimental results were simulated using the program "KINETIC 3.1" for Windows 3.x. The MDL 73,404 was oxidized by horseradish peroxidase in the presence of H2O2 to its corresponding MDL 73,404 quinone. During this oxidation, the horseradish peroxidase showed an unexpectedly slow kinetic response with time, which contrast with the linear product accumulation curve measured with 2,2'-azino-bis-(3-estilbenzotiazol-6-sulfonic acid) (ABTS). This response was dependent on the respective concentrations of enzyme, MDL 73,404 and H2O2. However, when the enzyme was incubated with H2O2, the slow kinetic response disappeared and a lag period was observed. Furthermore, when p-coumaric acid (PCA) was added, the activity increased and the slow kinetic response became a straight line. In order to explain this anomalous behaviour, a kinetic model has been proposed and its differential equations simulated. From the correlation between experimental and simulated results it is concluded that MDL 73,404 can act as a slow response substrate for peroxidase, probably due to the presence of a quaternary ammonium side chain that confers on it a slow capacity to convert compound III into ferriperoxidase.

Coumaric Acids↗