Thermal Inactivation Of Polyphenol Oxidase Research Articles

Comparison of the biochemical properties and thermal inactivation of polyphenol oxidase from three lily bulb cultivars.

The biochemical properties and thermal inactivation of polyphenol oxidase (PPO) from three main planted lily cultivars in China, namely, Lilium lancifolium Thunb, Lilium brownie var. viridulum, and Lilium davidii var. unicolor cotton were evaluated. Data indicate that the PPO from three cultivars showed two optimum pH levels of 4.0 and 6.5-7.0 and temperature of 15°C and exhibited the highest affinity toward 4-methylcatechol. However, this enzyme did not exhibit monophenolase activity. Thiourea and L-cysteine were more effective than other inhibitors. The enzymatic activity of L. lancifolium Thunb PPO crude extract was higher than that of L. brownie var. viridulum and L. davidii var. unicolor cotton. For thermal inactivation, L. davidii var. unicolor cotton PPO showed the best thermal resistance at 65-75°C, and L. lancifolium Thunb showed stability at 45°C. The deactivation of the three types of PPO followed the first-order reaction kinetics, and the activation energy (Ea) was 144.28, 138.00, and 107.12kJ/mol for L. lancifolium Thunb PPO, L. brownie var. viridulum PPO, and L. davidii var. unicolor cotton PPO, respectively. PRACTICAL APPLICATIONS: Lilium is an ornamental and edible plant typically used for food and traditional Chinese medicine. Its flowers are used for decoration, and its underground bulbs are rich in various bioactive substances. Fresh lily bulbs easily turn brown and lose economic value during storage and processing. Polyphenol oxidase (PPO) is a crucial molecule involved in the enzymatic browning of fruit and vegetables. In this study, PPO was extracted from three main planted lily cultivars in China. Namely, Lilium lancifolium Thunb, Lilium brownie var. viridulum, Lilium davidii var. unicolor cotton and was partially characterized. The results are of considerable importance to further understand the PPO of lily bulbs and provide guidance for the inactivation of enzymes and the processing of lily bulb juice.

Thermal inactivation of polyphenoloxidase and peroxidase in Jubileu clingstone peach and yeast isolated from its spoiled puree

The thermal inactivation of yeast isolated from spoiled Jubileu peach puree and that of polyphenoloxidase (PPO) and peroxidase (POD) in cv. Jubileu, which is widely cultivated in southern Rio Grande do Sul state, Brazil, were studied. PPO and POD were extracted using the protein powder method and submitted to partial purification by precipitation followed by dialysis. The enzymatic activity was determined measuring the increase in absorbance at 420 nm for PPO and 470 nm for POD. The yeast used in this investigation was isolated from spoiled Jubileu peach puree at 22 °Brix, with total initial microbial count of 22 × 10² UFCmL- 1. Stock cultures were maintained on potato dextrose agar (PDA) slants at 4 °C and pH 5 for later use for microbial growth. In all cases, kinetic analysis of the results suggests that the thermal inactivation was well described by a first-order kinetic model, and the temperature dependence was significantly represented by the Arrhenius law. Both enzymes were affected by heat denaturation, and PPO was more thermostable. PPO was also more thermosTable than the yeast isolated from peach puree. The D60-values were 1.53 and 1.87 min for PPO and yeast isolated from spoiled Jubileu peach puree, respectively.

Kinetics and Thermodynamics of the Thermal Inactivation of Polyphenol Oxidase in an Aqueous Extract from Agaricus bisporus

The kinetics and thermodynamics of the thermal inactivation of polyphenol oxidase (PPO) in an aqueous extract from mushroom Agaricus bisporus (J.E. Lange) Imbach was studied, using pyrocatechol as a substrate. Optimal conditions for enzymatic studies were determined to be pH 7.0 and 35-40 °C. The kinetics of PPO-catalyzed oxidation of pyrocatechol followed the Haldane model with an optimum substrate concentration of 20 mM. Thermal inactivation of PPO was examined in more detail between 50 and 73 °C and in relation to exposure time. Obtained monophasic kinetics were adequately described by a first-order model, with significant inactivation occurring with increasing temperature (less than 10% preserved activity after 6 min at 65 °C). Arrhenius plot determination and calculated thermodynamic parameters suggest that the PPO in aqueous extract from Agaricus bisporus mushroom is a structurally robust yet temperature-sensitive biocatalyst whose inactivation process is mainly entropy-driven.

Inactivation kinetics of polyphenol oxidase and peroxidase in green coconut water by microwave processing

The inactivation kinetics of enzymes polyphenol oxidase (PPO) and peroxidase (POD) was studied for the batch (discontinuous) microwave treatment of green coconut water. Inactivation of commercial PPO and POD added to sterile coconut water was also investigated. The complete time–temperature profiles of the experimental runs were used for determination of the kinetic parameters D-value and z-value: PPO ( D 92.20 °C = 52 s and z = 17.6 °C); POD ( D 92.92 °C = 16 s and z = 11.5 °C); PPO/sterile coconut water: ( D 84.45 °C = 43 s and z = 39.5 °C) and POD/sterile coconut water: ( D 86.54 °C = 20 s and z = 19.3 °C). All data were well fitted by a first order kinetic model. The enzymes naturally present in coconut water showed a higher resistance when compared to those added to the sterilized medium or other simulated solutions reported in the literature. The thermal inactivation of PPO and POD during microwave processing of green coconut water was significantly faster in comparison with conventional processes reported in the literature.

Partial characterization of lettuce (Lactuca sativa L.) polyphenol oxidase

Polyphenol oxidase (PPO) from garden lettuce (Lactuca sativa L.) was partially purified by ammonium sulphate ((NH4)2SO4) precipitation and dialysis, and then some of its kinetic properties such as optimum pH and temperature, substrate specificity, thermal inactivation and inhibition were investigated. The total phenolic and protein contents of Lactuca sativa L. extracts were determined according to the Folin-Ciocalteu and Bradford methods, and found to be 304 mg/100 g on a fresh weight basis and 494 μg/mL, respectively. PPO activity was determined using 4-methylcatechol, catechol and pyrogallol as substrates. Kinetic parameters, Km and Vmax, were calculated from Lineweaver–Burk plots. According to Vmax/Km ratio, pyrogallol was the most suitable substrate, followed by catechol and 4-methylcatechol. The optimum temperature and pH values were 30, 40 and 30 °C; and 6.5, 8.0 and 7.5 for 4-methylcatechol, catechol and pyrogallol substrates, respectively. The thermal inactivation of PPO was investigated at 35, 55 and 75 °C. The enzyme activity decreased with increasing temperature. The effect of different inhibitors on partially purified Lactuca sativa L. PPO was spectrophotometrically investigated. For this purpose, tropolone, glutathione, ascorbic acid and 4-aminobenzoic acid were used to inhibit the activity of Lactuca sativa L. PPO at different concentrations. From the experimental results, it was found that glutathione was found to be the most potent inhibitor for Lactuca sativa L. PPO.

THERMAL AND HIGH PRESSURE INACTIVATION KINETICS OF VICTORIA GRAPE POLYPHENOL OXIDASE: FROM MODEL SYSTEMS TO GRAPE MUST

ABSTRACT Polyphenol oxidase (PPO) was extracted from white grapes (var. Victoria, South Africa) and partially purified by ammonium sulfate precipitation. Detailed kinetic studies on thermal and high‐pressure inactivation of grape PPO were preformed in McIlvaine buffer (pH = 4.0), in reconstituted grape must and in grape must. Thermal inactivation of PPO in McIlvaine buffer followed a first‐order model while in reconstituted grape must and grape must a biphasic inactivation behavior has been observed. Under all conditions investigated, a first‐order kinetic model could describe the pressure inactivation data. It was found that PPO is more thermostable and pressure‐stable in must than in reconstituted grape must and McIlvaine buffer at the same pH = 4.0.

Thermal inactivation of polyphenoloxidase in pineapple puree

Prevention of browning in pineapple puree by thermal inactivation of enzyme, Polyphenoloxisase (PPO), was examined between 40 and 90 °C and in relation to exposure time. The amount of inactivation was measured as a function of time and temperature under isothermal conditions. Reaction rate constant and activation energy ( E a) as well as Decimal reduction time ( D) and z-value of thermal inactivation, were determined. The rate of inactivation varied with temperatures and follows a logarithmic law. Kinetic studies showed that the thermal inactivation (40–90 °C) of the PPO followed first-order kinetics.