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1.
Food Chem ; 343: 128401, 2021 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-33228966

RESUMEN

Enzymatic browning is one of the major difficulties for the preservation and commercial value of fresh-cut products. To research more healthy and inexpensive anti-browning methods, we investigated the effect of ultrasonic coupling purslane extract on the browning resistance of fresh-cut potato during 8d storage at 4 °C. Firstly, the optimal ultrasonic time (10 min) was obtained. Then, the results showed that the combined application with lower purslane extract concentration (0.02%, w/w) could achieve a better anti-browning effect than the optimal concentration of alone purslane extract (0.05%, w/w). The combined application not only significantly inhibited the key enzyme activities of polyphenol oxidase (PPO) and peroxidase (POD), but also effectively reduced the damage to cell membrane, maintained its integrity and permeability. Meanwhile, it also improved antioxidant capacity during storage. Overall, the ultrasonic cavitation combined with purslane extract would be a promising method for fresh-cut industry.


Asunto(s)
Almacenamiento de Alimentos/métodos , Extractos Vegetales/química , Portulaca/química , Solanum tuberosum/metabolismo , Ultrasonido , Antioxidantes/química , Catecol Oxidasa/antagonistas & inhibidores , Catecol Oxidasa/metabolismo , Color , Conductividad Eléctrica , Permeabilidad/efectos de los fármacos , Peroxidasa/antagonistas & inhibidores , Peroxidasa/metabolismo , Extractos Vegetales/metabolismo , Extractos Vegetales/farmacología , Portulaca/metabolismo , Solanum tuberosum/química , Solanum tuberosum/efectos de los fármacos
2.
Bioengineered ; 11(1): 281-290, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-32125230

RESUMEN

In this study, the conditions of extraction of loquat flowers polyphenolics were optimized through response surface methodology (RSM). Proper extraction conditions were: solid to liquid ratio 1 g per 50 mL and ethanol concentration 50% at 61°C for 9 min. Furthermore, the antioxidant and anti-polyphenol oxidase (PPO) activity of purified total polyphenolics (PTP) were investigated. PTP displayed strong antioxidant activity with IC50 values of 126.3 ± 8.9, 162.4 ± 6.3 and 94.97 mg ascorbic acid equivalent/g dry weight (mg AAE/d.w.) for ABTS, DPPH, and FRAP assays. In addition, PTP has a substantial inhibitory activity on PPO (IC50 = 115 ± 9.2 µg/mL). From the kinetics analysis, it was proved to be a reversible and mixed-type inhibitor of PPO with KI and KIS values of 76.77 µg/mL and 227.86 µg/mL, respectively. Further, the molecular mechanism underlying the inhibition of PPO by PTP was investigated by molecular docking techniques. The results showed that PTP units could form interaction with the catalytic pocket of PPO through the interaction with amino acid residues in the enzyme active center. The antioxidant activities of PTP together with its effect on PPO activity provide a strong starting point for their practical usage in the food industry.


Asunto(s)
Antioxidantes/química , Catecol Oxidasa/antagonistas & inhibidores , Eriobotrya/química , Flores/química , Extractos Vegetales/química , Etanol/química , Cinética , Oxidación-Reducción/efectos de los fármacos , Extractos Vegetales/farmacología
3.
Protein Expr Purif ; 171: 105612, 2020 07.
Artículo en Inglés | MEDLINE | ID: mdl-32097726

RESUMEN

In the literature, the polyphenol oxidase (PPO) enzyme has been purified a many times via Sepharose 4B-l-tyrosine-p-aminobenzoic acid affinity column. In order to study PPO purification efficiency, 2-aminophenol and 4-aminophenol were applied as a spacer arm to CNBr-activated Sepharose 4B. The effects of the spacer arm on specific activity, purification fold, and electrophoretic properties were investigated. The best performance with 11.7-fold purification and 23951 U/mg protein specific activity was achieved with the 4-aminophenol extension arm. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with was done to check the purity of the potato PPO enzyme obtained from affinity columns. According to the results of SDS-PAGE and native PAGE, the molecular weight of the enzyme is 50 kDa. Furthermore, the inhibition effects of curcumin and quercetin on the enzyme activity were examined, and the IC50 and Ki values were computed for the mentioned substances. IC50 values were determined to be 0.018 and 0.029 mM for potato PPO with curcumin and quercetin inhibitors with catechol as a substrate, respectively. IC50 value was also determined to be 0.0086 mM for quercetin inhibitor with 4-methylcatechol as a substrate. Ki constant was 0.0753 ± 0.0085 mM for curcumin using catechol as a substrate. No inhibition effect was observed for curcumin with the 4-methylcatechol substrate. The Ki constant for quercetin was 0.0398 ± 0.00743 mM with the 4-methylcatechol substrate and 0.0109 ± 0.0021 mM with the catechol substrate.


Asunto(s)
Catecol Oxidasa , Curcumina/química , Proteínas de Plantas , Tubérculos de la Planta/enzimología , Quercetina/química , Solanum tuberosum/enzimología , Catecol Oxidasa/antagonistas & inhibidores , Catecol Oxidasa/química , Catecol Oxidasa/aislamiento & purificación , Proteínas de Plantas/antagonistas & inhibidores , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación
4.
J Agric Food Chem ; 68(8): 2467-2476, 2020 Feb 26.
Artículo en Inglés | MEDLINE | ID: mdl-32031791

RESUMEN

Enzymatic browning is a major issue affecting the quality of processed potato (Solanum tuberosum L.). To understand the molecular mechanism of browning, transcriptional analyses were performed by employing potatoes that differed in browning. Coexpression analysis indicated that 9 out of 15 upregulated genes in browning-less groups encoded for potato protease inhibitors (StPIs). In addition, gene otology analysis showed that the enriched terms were mainly involved in protease inhibitors. Overexpression of cysteine StPI 143 and StPI 146 individually reduced browning and lowered protease activities and tyrosine and total free amino acid (FAA) contents, but they could not decrease polyphenol oxidase activity. Moreover, supplementing exogenous tyrosine or total FAAs into transgenic potato mash to wild-type amounts promoted mash browning, browning with total FAAs, more than with tyrosine, resembling wild-type levels. These results implied that cysteine StPIs reduced browning via lowering the accumulation of FAAs in addition to tyrosine. Our findings have enriched the knowledge about the roles and mechanisms of protease inhibitors in regulating enzymatic browning of potato, which provide new ways for controlling potato browning.


Asunto(s)
Aminoácidos/metabolismo , Proteínas de Plantas/metabolismo , Inhibidores de Proteasas/metabolismo , Solanum tuberosum/metabolismo , Catecol Oxidasa/antagonistas & inhibidores , Catecol Oxidasa/genética , Catecol Oxidasa/metabolismo , Color , Proteasas de Cisteína/genética , Proteasas de Cisteína/metabolismo , Proteínas de Plantas/genética , Solanum tuberosum/enzimología , Solanum tuberosum/genética
5.
J Sci Food Agric ; 100(5): 2065-2073, 2020 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-31875969

RESUMEN

BACKGROUND: Polyphenol oxidase (PPO) is considered a problem in the food industry because it starts browning reactions during fruit and vegetable processing. Ultrasonic treatment is a technology used to inactivate the enzyme; however, the mechanism behind PPO inactivation is still unclear. For this reason, the inactivation, aggregation, and structural changes in PPO from quince juice subjected to ultrasonic treatments were investigated. Different intensities and times of ultrasonic treatment were used. Changes in the activity, aggregation, conformation, and structure of PPO were investigated through different structural analyses. RESULTS: Compared to untreated juice, the PPO activity in treated juice was reduced to 35% at a high ultrasonic intensity of 400 W for 20 min. The structure of PPO determined from particle size distribution (PSD) analysis showed that ultrasound treatment caused initial dissociation and subsequent aggregation leading to structural modification. The spectra of circular dichroism (CD) analysis of ultrasonic treated PPO protein showed a significant loss of α-helix, and reorganization of secondary structure. Fluorescence analysis showed a significant increase in fluorescence intensity of PPO after ultrasound treatment with evident blue shift, revealing disruption in the tertiary structure. CONCLUSION: In summary, ultrasonic treatment triggered protein aggregation, distortion of tertiary structure, and loss of α-helix conformation of secondary structure causing inactivation of the PPO enzyme. Hence, ultrasound processing at high intensity and duration could cause the inactivation of the PPO enzyme by inducing aggregation and structural modifications. © 2019 Society of Chemical Industry.


Asunto(s)
Catecol Oxidasa/metabolismo , Jugos de Frutas y Vegetales/análisis , Ultrasonido , Catecol Oxidasa/antagonistas & inhibidores , Fenómenos Químicos , Dicroismo Circular , Color , Manipulación de Alimentos , Frutas/química , Calor , Concentración de Iones de Hidrógeno , Reacción de Maillard , Tamaño de la Partícula , Proteínas de Plantas/metabolismo , Estructura Secundaria de Proteína , Rosaceae/química , Verduras/química
6.
Food Chem ; 298: 125004, 2019 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-31260986

RESUMEN

There is general interest in strategies to control polyphenol oxidase (PPO)-initiated enzymatic browning because it is often associated with declining food quality. Cyclodextrins are cyclic glucan oligosaccharides that form inclusion complexes with a number of PPO substrates. This study focuses on the effect of ß-cyclodextrins (ßCyD) on PPO-catalyzed reactions. Potato enzyme extracts and semi-purified potato PPO served as enzyme sources. Substrates included phenolics endogenous to potatoes. Reaction time-courses were followed spectrophotometrically; rates were compared by analysis of variance. Extents of ßCyD inhibition of PPO-catalyzed reactions are shown to be substrate specific and can be quantitatively accounted for based on degrees of ßCyD substrate sequestration. There was no evidence for direct irreversible ßCyD inactivation of potato PPO. An apparent "direct PPO inactivation" by ßCyD is shown to result from a sequence of sequestration-dependent reactions that occur in commonly employed assay systems for the quantification of PPO in fruits and vegetables.


Asunto(s)
Catecol Oxidasa/química , Catecol Oxidasa/metabolismo , Solanum tuberosum/enzimología , beta-Ciclodextrinas/química , Catálisis , Catecol Oxidasa/antagonistas & inhibidores , Fenoles/química , Fenoles/metabolismo , Proteínas de Plantas/antagonistas & inhibidores , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Especificidad por Sustrato , beta-Ciclodextrinas/metabolismo
7.
J Food Sci ; 84(7): 1730-1736, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-31218701

RESUMEN

With high phytochemical and starch contents, purple-fleshed sweetpotatoes (PFSP) have been processed into various functional ingredients and food products including juices and natural colorants. For juice processing, PFSP are usually subjected to heat treatment for inactivation of pigment-degrading enzymes. However, heating of sweetpotatoes gelatinizes starch and produces thick slurry with cooked flavor, which are the drawbacks. Development of alternative processes to overcome the stated problems will be beneficial to sweetpotato processors. This study demonstrated that acidified water (≥3% w/v citric acid) was effective in inhibiting polyphenol oxidase and peroxidase in raw PFSP resulting in an attractive reddish juice. About 93% total phenolics (TP) and 83% total monomeric anthocyanins (TMA) in PFSP were extracted by two repeated extractions. The combined PFSP juice (3.2 L/kg PFSP) had high levels of TP (1,850 mg/L) and TMA (475 mg/L). With the developed process, 167 g dried starch, and 140 g dried high-fiber pomace were obtained for each kg raw PFSP, besides the highly pigmented juice. Pasteurization of the PFSP juice samples (pH 3.2) at 80 °C for 12 s resulted in 15% loss in TMA and had no effect on TP. The results indicated an efficient process to produce sweetpotato juice with high bioactive compounds and recovery of starch and high dietary fiber pomace as co-products. PRACTICAL APPLICATION: Purple-fleshed sweetpotatoes (PFSP) are rich in polyphenolics and antioxidant activities. In PFSP juice extraction, heat treatment to inactivate the pigment-degrading enzymes results in starch gelatinization and cooked flavor. A nonthermal process using acidified water was developed for producing anthocyanin-rich juice from PFSP and concurrently recovering native starch and dried pomace, which would increase the economic feasibility of the developed process. The results demonstrate an efficient process for the sweetpotato industry in producing PFSP pigmented juice and co-products for various food applications.


Asunto(s)
Ácidos/química , Antocianinas/análisis , Catecol Oxidasa/antagonistas & inhibidores , Jugos de Frutas y Vegetales/análisis , Ipomoea batatas/química , Peroxidasa/antagonistas & inhibidores , Extractos Vegetales/análisis , Proteínas de Plantas/antagonistas & inhibidores , Antocianinas/aislamiento & purificación , Catecol Oxidasa/análisis , Color , Culinaria , Fibras de la Dieta/análisis , Ipomoea batatas/enzimología , Peroxidasa/análisis , Fenoles/análisis , Fenoles/aislamiento & purificación , Extractos Vegetales/aislamiento & purificación , Proteínas de Plantas/análisis , Almidón/análisis
8.
Food Chem ; 283: 445-453, 2019 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-30722896

RESUMEN

Enzymatic browning is a crucial reaction affecting the quality of fresh-cut fruit and vegetables. Purslane is an edible Chinese folk medicine with extensive distribution and containing a lot of polyphenols and alkaloids. However, little research on its' anti-browning effect on fresh-cut food was reported. In this study, the effectiveness of 0.05% (w/w) purslane aqueous extract treatment efficiently inhibited the activities of polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonia-lyase (PAL), the membrane integrity was effectively maintained, and malondialdehyde (MDA) content increases was retarded during whole storage period at 4 °C. Oddly, the higher purslane extract concentration, the lower endogenesis phenolic content. Additionally, thirty polyphenols and fifty-six alkaloids were found in purslane aqueous extract by LC-MS/MS. All results suggest that purslane aqueous extract is a promising nutritive anti-browning agent for fresh-cut potato.


Asunto(s)
Extractos Vegetales/química , Portulaca/química , Solanum tuberosum/química , Alcaloides/análisis , Alcaloides/química , Alcaloides/metabolismo , Catecol Oxidasa/antagonistas & inhibidores , Catecol Oxidasa/metabolismo , Cromatografía Líquida de Alta Presión , Almacenamiento de Alimentos , Malondialdehído/metabolismo , Peroxidasa/antagonistas & inhibidores , Peroxidasa/metabolismo , Fenilanina Amoníaco-Liasa/antagonistas & inhibidores , Fenilanina Amoníaco-Liasa/metabolismo , Extractos Vegetales/metabolismo , Polifenoles/análisis , Polifenoles/química , Polifenoles/metabolismo , Portulaca/metabolismo , Solanum tuberosum/metabolismo , Espectrometría de Masas en Tándem , Agua/química
9.
Food Chem ; 274: 345-350, 2019 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-30372949

RESUMEN

Although enzymatic browning is important for the beneficial coloration of some foods, it also causes negative effects on safety, quality, and nutritional values of fruit and vegetable. Thus, anti-browning natural compounds have gained attention in the food industry. Xanthone-related compounds have been well-known for its biological activities, but their roles in enzymatic browning are unclear. We screened xanthone-related natural compounds for their anti-polyphenol oxidase (PPO) activity and maclurin was selected for further experiments due to stronger PPO-inhibitory activity. Maclurin suppressed enzymatic browning in potato supernatant for long-term partly through direct binding to and inactivating PPO presumably by forming multiple hydrogen bonds and aromatic interactions with the binding pocket. In addition, maclurin elevated antioxidant capacity when added to potato supernatant. Considering the diverse health-promoting effects of antioxidants, maclurin can be applied as a functional food additive to block enzymatic browning and increase the antioxidant property of foods including beverages and soups.


Asunto(s)
Antioxidantes/química , Antioxidantes/farmacología , Catecol Oxidasa/metabolismo , Aditivos Alimentarios/química , Aditivos Alimentarios/farmacología , Xantonas/química , Xantonas/farmacología , Catecol Oxidasa/antagonistas & inhibidores , Color , Oxidación-Reducción/efectos de los fármacos , Solanum tuberosum/efectos de los fármacos , Solanum tuberosum/metabolismo
10.
Comput Biol Chem ; 68: 131-142, 2017 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-28340400

RESUMEN

Polyphenol oxidases (PPOs)/tyrosinases are metal-dependent enzymes and known as important targets for melanogenesis. Although considerable attempts have been conducted to control the melanin-associated diseases by using various inhibitors. However, the exploration of the best anti-melanin inhibitor without side effect still remains a challenge in drug discovery. In present study, protein structure prediction, ligand-based pharmacophore modeling, virtual screening, molecular docking and dynamic simulation study were used to screen the strong novel inhibitor to cure melanogenesis. The 3D structures of PPO1 and PPO2 were built through homology modeling, while the 3D crystal structures of PPO3 and PPO4 were retrieved from PDB. Pharmacophore modeling was performed using LigandScout 3.1 software and top five models were selected to screen the libraries (2601 of Aurora and 727, 842 of ZINC). Top 10 hit compounds (C1-10) were short-listed having strong binding affinities for PPO1-4. Drug and synthetic accessibility (SA) scores along with absorption, distribution, metabolism, excretion and toxicity (ADMET) assessment were employed to scrutinize the best lead hit. C4 (name) hit showed the best predicted SA score (5.75), ADMET properties and drug-likeness behavior among the short-listed compounds. Furthermore, docking simulations were performed to check the binding affinity of C1-C10 compounds against target proteins (PPOs). The binding affinity values of complex between C4 and PPOs were higher than those of other complexes (-11.70, -12.1, -9.90 and -11.20kcal/mol with PPO1, PPO2, PPO3, or PPO4, respectively). From comparative docking energy and binding analyses, PPO2 may be considered as better target for melanogenesis than others. The potential binding modes of C4, C8 and C10 against PPO2 were explored using molecular dynamics simulations. The root mean square deviation and fluctuation (RMSD/RMSF) graphs results depict the significance of C4 over the other compounds. Overall, bioactivity and ligand efficiency profiles suggested that the proposed hit may be more effective inhibitors for melanogenesis.


Asunto(s)
Catecol Oxidasa/antagonistas & inhibidores , Evaluación Preclínica de Medicamentos , Inhibidores Enzimáticos/farmacología , Simulación de Dinámica Molecular , Catecol Oxidasa/metabolismo , Inhibidores Enzimáticos/química , Humanos , Simulación del Acoplamiento Molecular , Estructura Molecular , Relación Estructura-Actividad
11.
J Complement Integr Med ; 14(1)2017 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-28195547

RESUMEN

Background Phenolic compounds from Citrus are known to be a topic of many studies due to their biological properties including antioxidant activity. Methods Methanolic and aqueous extracts were isolated from Citrus leaves of different species (C. clementina, C. limon, C. hamlin, C. navel, C. aurantifolia, C. aurantium and C. grandis) harvested in Algeria. Results The results showed that aqueous extracts of all species are rich in total phenolic compounds and flavonoids (from 68.23 to 125.28 mg GAE/g DM) and (from 11.99 to 46.25 mg QE/g DM) respectively. The methanolic and aqueous extracts were examined for in vitro antioxidant properties using various antioxidant assays. For aqueous extracts, C. limon showed an important DPPH radical scavenging activity (IC50 35.35 µg/mL), and C. clementina exerted the highest ABTS radical scavenging activity (1,174.43 µM ET/g DM) and a significant ferric reducing potential (30.60 mg BHAE/g DM). For methanolic extracts, C. clementina showed the highest antioxidant activity for all the realized assays (IC50 41.85 µg/mL, 378.63 µM ET/g DM and 13.85 mg BHAE/g DM) for DPPH, ABTS radicals scavenging activities and ferric reducing potential respectively. Antiperoxidase and antipolyphenol oxidase activities of these samples were also evaluated. Conclusions In this investigation, the assessment of antiperoxidase activity proved that the leaves extracts of different species were able to inhibit peroxidase activity. However, this inhibition varied with the species and the source of these enzymes. On the other hand, the aqueous extracts of different species showed moderate inhibition of polyphenol oxidase, while no effect on these enzymes was obtained with methanolic extracts.


Asunto(s)
Antioxidantes/farmacología , Citrus/química , Flavonoides/farmacología , Reacción de Maillard , Oxidorreductasas/antagonistas & inhibidores , Fenoles/farmacología , Extractos Vegetales/farmacología , Argelia , Benzotiazoles/metabolismo , Compuestos de Bifenilo/metabolismo , Catecol Oxidasa/antagonistas & inhibidores , Peroxidasas/antagonistas & inhibidores , Picratos/metabolismo , Hojas de la Planta/química , Especificidad de la Especie , Ácidos Sulfónicos/metabolismo
12.
J Sci Food Agric ; 97(5): 1420-1426, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-27378649

RESUMEN

BACKGROUND: Enzymatic browning of fruits and vegetables and their products is an important factor worsening their quality. The influence of five green tea extracts at the concentrations of 1 g L-1 , 2 g L-1 and 3 g L-1 on polyphenol oxidase (PPO) activity in fresh cloudy apple juice was investigated. Moreover, PPO inhibition by tea extract and colour stability of juice during short-time refrigerated storage was studied. The changes of juice colour during storage was expressed as the total colour differences (ΔE*), browning index (BI), yellowness index (YI), and the absorbance at 420 nm (A420 ). RESULTS: All extracts inhibited PPO activity in fresh apple juice in concentration-dependent manner. PPO activity in pure apple juice decreased by 7% after 48 h, whereas PPO activity in samples with 1 g L-1 , 2 g L-1 and 3 g L-1 tea extract decreased by 53%, 74%, and 96%, respectively. Browning of apple juice during storage decreased with increased concentration of green tea extract. After 48 h, extract at 1 g L-1 , 2 g L-1 and 3 g L-1 inhibited browning of juice expressed as BI by 48%, 60%, and 86%, respectively, comparing to pure apple juice. CONCLUSION: Green tea extract may be an effective anti-browning agent for short-time stored cloudy apple juices. © 2016 Society of Chemical Industry.


Asunto(s)
Catecol Oxidasa/antagonistas & inhibidores , Manipulación de Alimentos/métodos , Jugos de Frutas y Vegetales/análisis , Malus/química , Extractos Vegetales , Té/química , Antioxidantes/análisis , Color , Almacenamiento de Alimentos , Malus/enzimología
13.
Food Chem ; 190: 922-927, 2016 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-26213057

RESUMEN

Full-fatted and commercially defatted rice bran extracts (RBE and CDRBE) were evaluated for their ability to inhibit enzymatic browning in potato and apple. RBE showed more effective inhibition of polyphenol oxidase (PPO) activity and browning in potato and apple as compared to CDRBE. Five phenolic compounds in RBE and CDRBE (protocatechuic acid, vanillic acid, p-coumaric acid, ferulic acid and sinapic acid) were identified by HPLC. They were then evaluated for their important role in the inhibition using a model system which found that ferulic acid in RBE and p-coumaric acid in CDRBE were active in enzymatic browning inhibition of potato and apple. p-Coumaric acid exhibited the highest inhibitory effect on potato and apple PPO (p ⩽ 0.05). Almost all phenolic compounds showed higher inhibitory effect on potato and apple PPO than 100 ppm citric acid.


Asunto(s)
Catecol Oxidasa/antagonistas & inhibidores , Reacción de Maillard , Malus/metabolismo , Oryza , Fenoles/farmacología , Extractos Vegetales/farmacología , Solanum tuberosum/metabolismo , Catecol Oxidasa/metabolismo , Oryza/química , Extractos Vegetales/análisis
14.
Molecules ; 20(12): 22422-34, 2015 Dec 14.
Artículo en Inglés | MEDLINE | ID: mdl-26694329

RESUMEN

Residual enzymatic activity in certain foods, particularly of polyphenoloxidase (PPO), is responsible for the majority of anthocyanin degradation in food systems, causing also parallel losses of other relevant nutrients. The present work explored the feasibility of modifying phenolic profiles of thyme extracts, by use of chromatographic resins, to obtain phenolic extracts capable of enhancing anthocyanin colour and stability in the presence of PPO activity. Results indicated that pretreatment of thyme extracts with strong-anion exchange resins (SAE) enhanced their copigmentation abilities with strawberry juice anthocyanins. Phenolic chromatographic profiles, by HPLC-PDA, also demonstrated that thyme extracts subjected to SAE treatments had significantly lower concentrations of certain phenolic compounds, but extracts retained their colour enhancing and anthocyanin stabilization capacities though copigmentation. Additional testing also indicated that SAE modified extract had a lower ability (73% decrease) to serve as PPO substrate, when compared to the unmodified extract. Phenolic profile modification process, reported herein, could be potentially used to manufacture modified anthocyanin-copigmentation food and cosmetic additives for colour-stabilizing applications with lower secondary degradation reactions in matrixes that contain PPO activity.


Asunto(s)
Antocianinas/química , Antioxidantes/química , Bebidas/análisis , Catecol Oxidasa/antagonistas & inhibidores , Fragaria/química , Thymus (Planta)/química , Resinas de Intercambio Aniónico/química , Antioxidantes/aislamiento & purificación , Color , Conservación de Alimentos/métodos , Humanos , Oxidación-Reducción , Fenoles/química , Extractos Vegetales/química
15.
Int J Biol Macromol ; 80: 469-74, 2015 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-26187193

RESUMEN

Polysaccharides differing in structure and chemical nature were screened for their ability to bind non-covalently with polyphenol oxidase (PPO) from potato (as a model) and their effect on enzyme activity. All the polysaccharides selected inhibited the PPO but ß-cyclodextrin showed maximum inhibition under optimum conditions. Process details for the inhibition of PPO were studied with respect to concentration of ß-cyclodextrin, temperature, pH, and time. Higher inhibition constant and lower half life was obtained at 40 °C than at 30 °C in the presence of inhibitor. ß-Cyclodextrin showed mixed type of inhibition of PPO. ß-Cyclodextrin was further exploited as anti-browning agent in selected fruit juices. It not only showed a significant anti-browning effect on freshly prepared potato juice but was also effective in other fruit juices. Better effect was seen in pineapple, apple and pear as compared to banana, sugarcane and guava fruit juices.


Asunto(s)
Catecol Oxidasa/química , Inhibidores Enzimáticos/química , Proteínas de Plantas/química , beta-Ciclodextrinas/química , Catecol Oxidasa/antagonistas & inhibidores , Estabilidad de Enzimas , Concentración de Iones de Hidrógeno , Cinética , Proteínas de Plantas/antagonistas & inhibidores , Unión Proteica , Solanum tuberosum/enzimología
16.
Food Chem ; 171: 382-7, 2015 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-25308684

RESUMEN

Crude extracts of Ataulfo exhibited polyphenol oxidase (PPO) activity with pyrogallol, 3-methylcatechol, catechol, gallic acid, and protocatechuic acid. The substrate dependent pH optima ranged from pH 5.4 to 6.4 with Michaelis-Menten constants between 0.84 ± 0.09 and 4.6 ± 0.7 mM measured in MES or phosphate buffers. The use of acetate buffers resulted in larger Michaelis-Menten constants, up to 14.62 ± 2.03 mM. Sodium ascorbate, glutathione, and kojic acid are promising inhibitors to prevent enzymatic browning in Ataulfo. PPO activity increased with ripeness and was always higher in the skin compared to the pulp. Sodium dodecyl sulphate (SDS) enhanced PPO activity, with pulp showing a stronger increase than skin. SDS-PAGE gels stained for catecholase activity showed multiple bands, with the most prominent bands at apparent molecular weights of 53, 112, and 144 kDa.


Asunto(s)
Catecol Oxidasa/metabolismo , Mangifera/enzimología , Ácido Ascórbico/química , Ácido Ascórbico/metabolismo , Catecol Oxidasa/antagonistas & inhibidores , Electroforesis en Gel de Poliacrilamida , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Glutatión/química , Glutatión/metabolismo , Cinética , Peso Molecular , Extractos Vegetales/metabolismo , Unión Proteica , Pironas/química , Pironas/metabolismo , Dodecil Sulfato de Sodio/química , Dodecil Sulfato de Sodio/metabolismo
17.
J Agric Food Chem ; 62(1): 214-21, 2014 Jan 08.
Artículo en Inglés | MEDLINE | ID: mdl-24344979

RESUMEN

Enzymatic browning is a major quality issue in fruit and vegetable processing and can be counteracted by different natural inhibitors. Often, model systems containing a single polyphenol oxidase (PPO) are used to screen for new inhibitors. To investigate the impact of the source of PPO on the outcome of such screening, this study compared the effect of 60 plant extracts on the activity of PPO from mushroom ( Agaricus bisporus , AbPPO) and PPO from potato ( Solanum tuberosum , StPPO). Some plant extracts had different effects on the two PPOs: an extract that inhibited one PPO could be an activator for the other. As an example of this, the mate ( Ilex paraguariensis ) extract was investigated in more detail. In the presence of mate extract, oxygen consumption by AbPPO was found to be reduced >5-fold compared to a control reaction, whereas that of StPPO was increased >9-fold. RP-UHPLC-MS analysis showed that the mate extract contained a mixture of phenolic compounds and saponins. Upon incubation of mate extract with StPPO, phenolic compounds disappeared completely and saponins remained. Flash chromatography was used to separate saponins and phenolic compounds. It was found that the phenolic fraction was mainly responsible for inhibition of AbPPO and activation of StPPO. Activation of StPPO was probably caused by activation of latent StPPO by chlorogenic acid quinones.


Asunto(s)
Agaricus/enzimología , Catecol Oxidasa/antagonistas & inhibidores , Catecol Oxidasa/metabolismo , Inhibidores Enzimáticos/química , Proteínas Fúngicas/antagonistas & inhibidores , Proteínas Fúngicas/metabolismo , Ilex paraguariensis/química , Extractos Vegetales/química , Proteínas de Plantas/antagonistas & inhibidores , Proteínas de Plantas/metabolismo , Solanum tuberosum/enzimología , Fenoles/química , Saponinas/química
18.
J Agric Food Chem ; 61(22): 5385-90, 2013 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-23683299

RESUMEN

Enzymatic browning by polyphenoloxidase (PPO) affects food quality and taste in fruits and vegetables. Thus, the study was designed to reduce browning in apple juice by coumarin. The ethanolic extract of cinnamon was prepared and its coumarin content was quantitated by HPLC, using authentic coumarin (AC) as standard. The effect of cinnamon extract (CE) and AC on enzymatic browning, its time dependent effects, and the specific activity of PPO and peroxidase (POD) were studied in apple juice. The docking of coumarin with PPO and POD was also performed to elucidate its antibrowning mechanism. The CE (73%) and AC (82%) showed better reduction in browning, maintained its antibrowning effect at all time points, and significantly (p < 0.05) reduced the specific activity of PPO and POD when compared with controls. Coumarin showed strong interaction with binding pockets of PPO and POD, suggesting its potential use as inhibitor to enzyme mediated browning in apple juice.


Asunto(s)
Bebidas/análisis , Cinnamomum zeylanicum/química , Cumarinas/química , Inhibidores Enzimáticos/química , Conservantes de Alimentos/química , Malus/química , Especias/análisis , Catecol Oxidasa/antagonistas & inhibidores , Catecol Oxidasa/química , Catecol Oxidasa/metabolismo , Color , Cumarinas/análisis , Cumarinas/aislamiento & purificación , Cumarinas/metabolismo , Inhibidores Enzimáticos/análisis , Inhibidores Enzimáticos/aislamiento & purificación , Inhibidores Enzimáticos/metabolismo , Conservantes de Alimentos/análisis , Conservantes de Alimentos/aislamiento & purificación , Conservantes de Alimentos/metabolismo , Calidad de los Alimentos , Frutas/química , Frutas/enzimología , Malus/enzimología , Conformación Molecular , Simulación del Acoplamiento Molecular , Peroxidasa/antagonistas & inhibidores , Peroxidasa/química , Peroxidasa/metabolismo , Corteza de la Planta/química , Extractos Vegetales , Proteínas de Plantas/antagonistas & inhibidores , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo
19.
Phytochemistry ; 90: 16-24, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23523329

RESUMEN

Ferula communis (L.), a plant belonging to Apiaceae, is widely present in Sardinia, Italy. Currently, interest in F. communis focuses on the presence of two chemotypes in the wild. One chemotype is poisonous to animals, whereas the other chemotype is non-poisonous. Polyphenol oxidase (PPO) has been extracted and partially purified from the two chemotypes of F. communis. The biochemical characterization of the enzymes showed significant differences. In particular, while the two PPOs were not able to use 6- and 7-hydroxycoumarin as substrates, they showed distinct specificity for 6,7- and 7,8-dihydroxycoumarin. Significant differences in the enzyme behavior towards common PPO inhibitors were also observed. In addition, activation energy and activation energy for denaturation were determined, showing significant differences between FP-PPO and FNP-PPO, particularly for denaturation kinetics. The possible roles of the two PPOs in determining differences in composition and toxicity of the two F. communis chemotypes are also discussed.


Asunto(s)
Catecol Oxidasa/aislamiento & purificación , Catecol Oxidasa/metabolismo , Ferula/enzimología , Catecol Oxidasa/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Concentración de Iones de Hidrógeno , Italia , Estructura Molecular , Relación Estructura-Actividad , Especificidad por Sustrato
20.
J Food Sci ; 78(3): C425-31, 2013 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-23425135

RESUMEN

UNLABELLED: Enzymatic browning remains a problem for the fruit and vegetable industry, especially new emerging markets like pre-cuts. A crude inhibitor from blue mussel (Mytilus edulis) showed broad inhibition for apple (58%), mushroom (32%), and potato (44%) polyphenol oxidase (PPO) and was further characterized. Inhibition increased as the concentration of inhibitor increased in the reaction mixture eventually leveling off at a maximum inhibition of 92% for apple PPO. The inhibitor was capable of bleaching the brown color formed in the reaction mixture with apple PPO. Identification of the inhibitor by mass spectrometry and high-performance liquid chromatography revealed it to be hypotaurine (C2 H7 NO2 S). Hypotaurine and other sulfinic acid analogs (methane and benzene sulfinic acids) showed very good inhibition for apple PPO at various concentrations with the highest inhibition occurring at 500 µM for hypotaurine (89%), methane sulfinic acid (100%), and benzene sulfinic acid (100%). PRACTICAL APPLICATION: An inhibitor found in the expressed liquid from blue mussel shows very good inhibition on enzymatic browning. Since this enzyme is responsible for losses to the fruit and vegetable industry, natural inhibitors that prevent browning would be valuable. Finding alternative chemistries that inhibit browning and understanding their mode of action would be beneficial to the fruit and vegetable industries and their segments such as pre-cuts, juices, and so on. Inhibitors from products ingested by consumers are more acceptable as natural ingredients.


Asunto(s)
Catecol Oxidasa/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/aislamiento & purificación , Mytilus edulis/química , Agaricales/enzimología , Animales , Cromatografía Líquida de Alta Presión , Frutas/enzimología , Malus/enzimología , Solanum tuberosum/enzimología , Verduras/enzimología
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