Kinetic Characterization and Thermal Inactivation of Peroxidase in Aqueous Extracts from Sweet Corn and Waxy Corn

The objective of this study was to evaluate the activity, kinetic behavior, and thermal inactivation kinetics of peroxidase (POD) in aqueous extracts from two kinds of milk ripe stage corn, sweet corn and waxy corn. Optimum activities using guaiacol as the hydrogen donor were obtained for sweet corn at pH 4.8 and for waxy corn at pH 6.0. The kinetics of POD showed characteristics which were dependent upon the concentrations of guaiacol and H₂O₂. The guaiacol K _m values for sweet corn POD and waxy corn POD were 11.01 and 23.01 mM, respectively, whereas the H₂O₂ K _m values for sweet corn POD and waxy corn POD were 2.85 and 0.33 mM, respectively. Thermal treatment of enzymatic aqueous extracts was carried out at different time–temperature combinations in the range of 0–25 min and 60–85 °C. Arrhenius plot determination and calculated thermodynamic parameters suggested that the inactivation of POD followed first-order reaction kinetics, and the activation energy (E _a) for inactivation of sweet corn POD (114.36 kJ/mol) was slightly lower compared with waxy corn POD (119.72 kJ/mol). There were several notable similarities between the inactivation kinetics in the two corn cultivars.     

Inactivation Kinetics of the Most Baro-Resistant Enzyme in High Pressure Processed Litchi-Based Mixed Fruit Beverage

This work focused on a litchi-based mixed fruit beverage, comprising of coconut water and lemon juice, mixed in an optimized proportion. Based on preliminary studies, three resistant spoilage enzymes were identified in the beverage, viz. polyphenol oxidase (PPO), peroxidase (POD), and pectin methyl esterase (PME). The response surface methodology (RSM) based on central composite face-centered design (FCCD) screened out PPO as the most resistant enzyme within the high pressure processing (HPP) domain of 200–600 MPa/30–70 °C/0–20 min. A detailed kinetic study was conducted on PPO inactivation within the same HPP domain along with a set of thermal treatments (0.1 MPa/30–70 °C). A synergistic effect of pressure and temperature on PPO inactivation was observed, throughout the HPP domain. However, PPO was almost completely inactivated at 500 MPa/70 °C/20 min. The inactivation order (n) values for PPO were 1.10 and 1.25 for thermal and HPP treatments, respectively. For every 10 °C rise in temperature, the inactivation rate constant (k, U^n-1 min^?1) increased approximately by 1.5 times, within 50–70 °C (at 0.1 MPa), while a 10-fold increase was obtained in the case of HPP treatments. The activation energy (E _a ) and the activation volume (V _a), depicting the temperature and pressure dependence of k, was found to decrease slightly, with an increase in pressure and temperature, respectively. The PPO inactivation rate constant was modeled as a function of both temperature and pressure conditions by combining both Arrhenius and Eyring equations.     

Pressure-Thermal Kinetics of Furan Formation in Selected Fruit and Vegetable Juices

Furan, a potential carcinogenic compound, can be formed in array of processed foods. The objective of this study was to conduct kinetic studies in pineapple juice and assess the interactive effects of pressure (0.1 to 600 MPa) and temperature (30 to 120 °C) on furan formation. Additional experiments were carried out in tomato, watermelon, cantaloupe, kale, and carrot juice to understand the influence of matrix and juice pH. Furan was monitored in raw (control) and processed samples by automated headspace gas chromatography mass spectrometry, and quantified by calibration curve method with d₄-furan as internal standard. The data were modeled using zero-, first-, and second-order equations. The zero-order rate constants (k _T,P ), activation energy (E _a ), and Gibbs free energy of activation (ΔG ^?) of furan formation in thermally processed (TP; 90–120 °C) pineapple juice were found to be 0.036–0.55 μg/kg/min, 98–114 kJ/mol, and 173.9–180.5 kJ/mol, respectively. Furan concentration was negligible and close to the detection limit (0.37 μg/kg) after pressure treatment (600 MPa at 30 °C) of juice samples. For similar process temperatures, the rate constants of pressure-assisted thermally processed (PATP; 600 MPa at 105 °C) pineapple juice were lower than that of TP samples. Furan formation was influenced by juice matrix and pH. On the other hand, PATP markedly suppressed furan (0.7 to 1.6 μg/kg) in these selected juices. In conclusion, furan formation increased with process temperature and treatment time, while pressure treatment at ambient temperature did not promote its production. Furan formation in TP fruit juices was also influenced by juice matrix and pH, but these were not the significant factors for PATP-treated juices.     

Kinetic characterisation and thermal inactivation study of polyphenol oxidase and peroxidase from table grape (Crimson Seedless)

Polyphenol oxidase (PPO) and peroxidase (POD) were extracted from a table grape (Crimson Seedless) using Triton X-114 and characterized using spectrophotometric methods. Both PPO and POD were activated by acid shock. However, in the presence of the anionic detergent sodium dodecil sulphate (SDS), PPO was activated whereas POD was inactivated. The enzymes were kinetically characterized and both followed Michaelis–Menten kinetics, although with different values of their kinetic parameters. The V_m/K_m ratio showed that Crimson Seedless grape PPO presents a similar affinity for 4-tert-butyl-catechol (TBC) whether activated by acid shock (0.018 min⁻¹) or SDS (0.023 min⁻¹). With regards to POD, the K_m and V_m values for 2,2′-azinobis(3-ethylbenzothiazolinesulphonic acid) (ABTS) were 0.79 mM and 1.20 μM/min, respectively. In the case of H₂O₂, the K_m and V_m value were 0.4 mM and 0.93 μM/min, respectively. PPO and POD showed similar thermostability, losing >90% of relative activity after only 5 min of incubation at 78 °C and 75 °C, respectively. In addition, PPO´s activation energy was similar to that obtained for POD (295.5 kJ/mol and 271.9 kJ/mol, respectively).     

Effect of pH on Enzyme Inactivation Kinetics in High-Pressure Processed Pineapple (Ananas comosus L.) Puree Using Response Surface Methodology

Effect of pH and high-pressure process treatments viz. pressure, temperature, and dwell time on inactivation of polyphenoloxidase (PPO), peroxidase (POD), bromelain (BRM), and pectinmethylesterase (PME) in pineapple puree was studied. Experiments were conducted according to rotatable central composite design (RCCD) within the range (?α to?+?α) of 100–600 MPa, 20–70 °C, and 0–30 min at three different pH levels (3.0, 3.5, and 4.0) followed by analysis through response surface methodology (RSM). Enzyme inactivation was significantly (p?k in min^?1) revealed that PPO was the most resistive (k ranged between 0.0020 and 0.0379 min^?1) when compared with other three enzymes within the experimental domain. Increased k at lower pH with constant pressure and temperature depicted that pH had negative effect on the inactivation process. The optimized conditions targeting maximum inactivation of PPO, POD and PME with simultaneous retention of BRM in pineapple puree, were 600 MPa/60 °C/9 min, 600 MPa/60 °C/10 min and 600 MPa/60 °C/10 min for the samples of pH 3.0, 3.5, and 4.0, respectively.     

Inactivation kinetics and secondary structural change of PEF-treated POD and PPO

Effects of pulsed electric fields (PEF) on the activity of peroxidase (POD) and polyphenol oxidase (PPO) in buffered solution were studied while the corresponding changes to their secondary structures was demonstrated by far-UV Circular dichroism (CD). The relative residual activity of POD and PPO decreased with the increase in electric field strength and treatment time, and PPO was more susceptible than POD to PEF treatment. The greatest reduction of the activity was achieved for POD at 25 kV/cm for 1740 μs and PPO at 25 kV/cm for 744 μs with reductions of 32.2% and 76.2%, respectively. The inactivation kinetic parameters D-value and Z_E value were calculated. The D-values of PPO were smaller than those POD at higher electric field strength, and Z_E values of POD and PPO were 36.9 and 16.2 kV/cm, respectively. The secondary structures of the two enzymes were changed following treatment by PEF. The intensity of negative peaks in the CD spectra decreased, and the CD spectra of PPO changed more significantly than that of POD; the reduction of the relative α-helix fractions for POD at 25 kV/cm for 124 μs was 22.63% while it was 50.72% for PPO at 25 kV/cm for 52 μs. The inactivation of PEF-treated POD and PPO was in close agreement with their secondary structure changes.     

Kinetics of Quality Changes of Shrimp (<Emphasis Type="Italic">Litopenaeus setiferus</Emphasis>) During Pasteurization

Effect of cooking between 1 and 80 min at 60 to 100 °C on several quality attributes of whole peeled shrimp (Litopenaeus setiferus) (80–90 counts/kg) was studied using an isothermal heating method. Cook loss, area shrinkage, and hardness of shrimp increased with increasing heating time and temperature, following a fractional first-order kinetic model with activation energies (E _a ) of 71.0, 53.3, and 29.9 kJ/mol, respectively. Cook loss, area shrinkage, and hardness were positively correlated. The toughness of shrimp muscle increased in the initial period of heating, then decreased in the later period during the treatments. The overall color change (ΔE) increased with increasing treatment time and temperature, and followed a zero kinetic model with an E _a of 37.2 kJ/mol. The kinetic parameters obtained from this study can be applied toward understanding and predicting shrimp-quality changes during pasteurization treatments, and further provides insight into the pasteurization conditions required to achieve safe and high-quality shrimp products and potentially other crustacean shellfish and seafood products.     

Thermal inactivation kinetics parameters of browning enzymes in starfruit (Averrhoa carambola L.) juice

This study aimed to evaluate the thermal inactivation kinetics of polyphenol oxidase (PPO) and peroxidase (POD) in starfruit juice. It followed the Malaysia Food Regulations 1985 and CODEX STAN 247-2005. Glucose, fructose and sucrose were the main sugars in starfruit juice. The total soluble solids, pH, titratable acidity, and total phenolics content of the starfruit juice produced were 8.13 ± 0.25 °Brix, 3.80 ± 0.05, 0.43% ± 0.02% malic acid, and 93.67 ± 4.96 mg GAEL⁻¹, respectively. Thermal inactivation kinetics of PPO and POD followed the first-order kinetic model. The decimal reduction time at 83.6 °C (D_83.6) of PPO and POD was 198.48 and 98.4 s, respectively, while the thermal resistance constant (z value) of PPO and POD was 12.8 and 5.4 °C, respectively. In conclusion, PPO might be a suitable signal for thermal processing on starfruit juice since it has higher heat resistance than POD.     

Biochemical characterization and process stability of polyphenoloxidase extracted from Victoria grape (Vitis vinifera ssp. Sativa)

Polyphenol oxidase (PPO) was isolated from Victoria grapes (Vitis vinifera ssp. Sativa) grown in South Africa and its biochemical characteristics were studied. Optimum pH and temperature for grape PPO activity were pH 5.0 and T = 25 °C with 10 mM catechol in McIlvaine buffer as substrate. PPO showed activity using the following substances: catechol, 4 methyl catechol, d, l-DOPA, (+) catechin and chlorogenic acid. K_m and V_max values were 52.6 ± 0.00436 mM and 653 ± 24.0 OD_400 nm/min in the case of 10 mM catechol as a substrate. Eight inhibitors were tested in this study and the most effective inhibitors were found to be ascorbic acid, l-cysteine and sodium metabisulfite. Kinetic studies showed that the thermal inactivation of Victoria grape PPO followed first-order kinetics, with an activation energy, E_a = 225 ± 13.5 of kJ/mol. Both in semipurified extract and in grape juice, PPO showed a pronounced high pressure stability.     

Thermal degradation kinetics of anthocyanins from Chinese red radish (Raphanus sativus L.) in various juice beverages

Thermal and storage stabilities of red radish anthocyanins (RRAs) in various juice beverages (apple, grape, peach, pear, pomegranate and lemon) were studied over temperature range 70–90 °C and 4–25 °C. RRAs degradation in all juice beverages followed first-order reaction kinetics. RRAs showed a much faster degradation rate during storage at room temperature (t _1/2 value ≤84.0 days) than did in refrigerated temperature (t _1/2 ≥value 130.9 days). The rate constant (k), E _a and Q ₁₀ values for RRAs in juice beverages varied from 1.33 to 0.33, 47.94 to 14.77 kJ mol^?1 and 1.16 to 1.89 at 70–90 °C. During heating, RRAs in peach and pomegranate showed higher stability than others at these temperatures. There was a positive correlation (R ² > 0.9128) between ascorbic acid content of juice beverages (8–36 mg/100 mg) and stability of RRAs at 70–90 °C. It was found that RRAs in apple and pear juice beverage were more stable than in other juice beverages.     

Characterization of polyphenoloxidase from 2 peach (Prunus persica L.) varieties grown in Argentina

Polyphenoloxidase was extracted from September peach (_SEPPO) and Summerset peach (_SUPPO) and its physicochemical characteristics were analyzed. The optimum pH was 6.5 for _SEPPO and 5.5 for _SUPPO. The optimum temperature was 35°C for _SEPPO and 39.4°C for _SUPPO. Activation energy (Ea) from thermal activation was 41.5 kJ/mol for _SEPPO and 37.5 kJ/mol for _SUPPO. Heating at 60°C by 5 min, _SUPPO was denatured whereas _SEPPO retained 2.6% of activity. Activation enthalpy (ΔH^#) and activation entropy (ΔS^#) for _SEPPO heat-inactivation were 69.9 J/mol and −83.5 kJ/mol·K for _SUPPO, ΔH^# was 91.8 J/mol while ΔS^# was −21.0 kJ/mol·K. Substrate specificity (V_max/K_M) was 4-methylcatechol>catechol>pyrogallol for _SEPPO and 4-mehtylcatechol>pyrogallol>catechol for _SUPPO. For both enzymes, the order of inhibition effectiveness using reductor agents was metabisulphite>ascorbic acid. Benzaldehyde, 4-hydroxybenzaldehyde, and dl-dopa were competitive inhibitors, and their K_I values were 38.86, 8.43, and 2.08 mM, respectively.     

Effect of High Hydrostatic Pressure and Temperature on Enzymatic Activity and Quality Attributes in Mango Puree Varieties (cv. Tommy Atkins and Manila)

Pectinmethylesterase (PME), peroxidase (POD), and polyphenoloxidase (PPO) residual activities (RAs) and physicochemical parameters (pH, total soluble solids (TSS), water activity (a_w), viscosity and color) of Tommy Atkins and Manila mango purees (MPs) were evaluated after high hydrostatic pressure (HHP) treatments at 400–550 MPa/0–16 min/34 and 59 °C. HHP treatment applied at 59 °C induced higher enzyme inactivation levels than the treatment applied at 34 °C in both MPs. The lowest RA of PME (26.9–38.6%) and POD (44.7–53%) was achieved in Manila MP treated at 450 MPa/8–16 min/59 °C and 550 MPa/4–16 min/59 °C, respectively. Otherwise, Tommy Atkins puree pressurized at 550 MPa/8–16 min/59 °C had the lowest PPO RA (28.4–34%). A slight decrease in pH and TSS values of both HHP-processed MPs at 34 and 59 °C was observed, whereas the a_w remained constant after processing. The viscosity of MPs tended to augment up to 2.1 times due to the application of HHP. No significant changes were observed in color parameters of Tommy Atkins MP, except at 550 MPa and 59 °C where higher yellow index (YI) (122.4?±?3.3) and lower L* (37.3?±?5.3) were obtained compared to the untreated MP. HHP caused an increase in L* values in Manila MP, whereas no clear trend was observed in YI. HHP processing at 550 MPa combined with mild temperature (59 °C) during 8 min could be a feasible treatment to reduce enzymatic activity and preserve fresh-like quality attributes in MP.     

Isolation, purification and determination of some biochemical properties of β-glucosidase from Muscat of Bornova grape

This research was undertaken to determine biochemical properties of β-glucosidase (β-d-glucoside glucohydrolase, EC 3.2.1.21) isolated from Muscat of Bornova grape. The optimum pH for β-glucosidase activity was found to be 5.0, and the enzyme showed high activity over a broad pH range of 4.5–6.0. However, due to low activity at pH 3.0, the enzyme is expected to exhibit only a fraction of the maximum activity during grape juice fermentation due to low pH of grape juice. As the temperature increased from 30 to 55 °C, the activity increased, too, the maximum activity occurring at 55 °C which implies that the enzyme is expected to exhibit a low activity at grape juice fermentation. According to thermal inactivation studies, k _D values increased as the temperature increased, whereas half-life and D values decreased. Energy of activation (E _a) and Z values were found to be 120.99 kj mol^?1 (r ² = 0.9776) and 18.08 °C (r ² = 0.9750), respectively. d-glucose and ethyl alcohol inhibited the enzyme at varying degrees depending on the concentration.     

Inactivation of peroxidase and polyphenol oxidase in red beet (Beta vulgaris L.) extract with continuous high pressure carbon dioxide

The inactivation of peroxidase (POD) and polyphenol oxidase (PPO) in red beet extract (RBE) with continuous high pressure carbon dioxide (HPCD) was investigated. HPCD treatment at 7.5 MPa (55 °C, 30 min) resulted in a reduction of their activities by approximately 73% and 93%, respectively. Compared with thermal treatment, continuous HPCD treatment reduced the decimal reduction time (D) of POD and PPO from 555.6 min to 55.9 min and 161.3 min to 32.1 min, respectively. The inactivation process could be described by first-order kinetics (r² > 0.70, p < 0.05); D values declined when temperature increased and continuous HPCD at 7.5 MPa and 55 °C resulted in the highest reaction rate constant (k value; smallest D value). The activation energy of the inactivation was reduced by HPCD treatment from 92.5 kJ/mol to 69.8 kJ/mol and 57.1 kJ/mol to 49.5 kJ/mol for POD and PPO, respectively. Continuous HPCD treatment had little effect on the antioxidant capacities of RBE samples.     

Kinetics of Thermal Inactivation of Peroxidase and Color Degradation of African Cowpea (Vigna unguiculata) Leaves

Cowpea leaves form an important part of the diet for many Kenyans, and they are normally consumed after a lengthy cooking process leading to the inactivation of peroxidase (POD) that could be used as an indicator for the potential shelf life of the vegetables. However, color degradation can simultaneously occur, leading to poor consumer acceptance of the product. The kinetics of POD in situ thermal (for thermal treatments in the range of 75 to 100 °C/120 min) inactivation showed a biphasic first‐order model, with Arrhenius temperature dependence of the rate constant. The kinetic parameters using a reference temperature (T_ref) of 80 °C were determined for both the heat‐labile phase (k_ref = 11.52 ± 0.95 × 10^?2 min^?1 and E_a of 109.67 ± 6.20 kJ/mol) and the heat‐stable isoenzyme fraction (k_ref = 0.29 ± 0.07 × 10^?2 min^?1 and E_a of 256.93 ± 15.27 kJ/mol). Color degradation (L*, a*, and b* value) during thermal treatment was investigated, in particular as the “a*” value (the value of green color). Thermal degradation (thermal treatments between 55 and 80 °C per 90 min) of the green color of the leaves followed a fractional conversion model and the temperature dependence of the inactivation rate constant can be described using the Arrhenius law. The kinetic parameters using a reference temperature (T_refC = 70 °C) were determined as k_refC = 13.53 ± 0.01 × 10^?2 min^?1 and E_aC = 88.78 ± 3.21 kJ/mol. The results indicate that severe inactivation of POD (as an indicator for improved shelf life of the cooked vegetables) is accompanied by severe color degradation and that conventional cooking methods (typically 10 min/100 °C) lead to a high residual POD activity suggesting a limited shelf life of the cooked vegetables.     

Determination of conformational changes of polyphenol oxidase and peroxidase in peach juice during mild heat treatment using FTIR spectroscopy coupled with chemometrics

The effect of thermal treatment on activity and structural changes of polyphenol oxidase (PPO) and peroxidase (POD) in peach juice was investigated. The D₇₀ values for PPO and POD were 10.38 and 13.68 min, respectively. The z values for PPO and POD were found to be 3.35 and 23.15 °C, respectively. Fourier transform infrared (FTIR) spectroscopy was used to determine the conformational changes. With the increase in temperature, α-helix and β-sheet were decreased, and β-turn and aggregated β-sheet structure were increased for PPO, random coil and β-sheet decreased, while aggregated β-sheet structure increased for POD. In contrast, an increase in α-helix structure of POD was observed which was thought to be effective in POD regeneration. Furthermore, native and denatured forms of enzymes were discriminated by using principle component analysis (PCA), and models that correlated activity and secondary structural components with infrared spectra were constructed by using partial least square (PLS).     

Study of the quality changes and myofibrillar proteins of white shrimp (Litopenaeus vannamei) under modified atmosphere packaging with varying CO2 levels

This paper studied the effect of modified atmosphere packaging (MAP) with 40–80 % CO₂, 5 % O₂ and 15–55 % N₂ on the quality of Pacific white shrimp (Litopenaeus vannamei) during 10-day storage at (4 ± 1) °C. The effect of CO₂ on the shrimp quality was compared by analyzing the quality indices such as total viable counts (TVC), total volatile basic nitrogen (TVB-N), pH and polyphenoloxidase (PPO) activity, whiteness and sensory attributes. Changes in myofibril length of shrimp flesh were also studied, which correspond to flesh softening. The findings suggest that the shrimp packaged in modified atmosphere had better quality indices than the control. Compared the MAP-batches, 80 % CO₂/15 % N₂/5 % O₂ led to the lowest level of TVC, TVB-N values, PPO activity and the highest sensory scores. Therefore, high-CO₂ packaging should be recommended to delay quality loss of shrimp during transportation and storage.     

Biochemical and Structural Changes of Taro (Colocasia esculenta) Tubers During Simple Thermal Treatments (Low Temperature) or in Combination with Chemicals

The present study set out to study the structural and biochemical modification of taro (Colocasia esculenta) grown in Cameroon, through simple heating or in association with chemicals. Both techniques are known to inactivate the reactions of polyphenol oxidase. The textural analysis was performed on tubers heated in water, 0.5% NaCl and 1% NaCl solution at various times from 0 to 105?min. The result showed optimum pH and temperature for taro polyphenol oxidase (PPO) activity at pH?6.0 and t?=?40?°C with 10?mM catechol in 0.1?M phosphate buffer as substrate. K _m and V _max values were about 7.317?±?0.012?mM and 0.148?±?0.0003 OD_{430 nm}/minute. Seven inhibitors were tested in this study, and the most effective inhibitors were found to be NaCl, CaCl2 and KCl. Kinetic studies showed that the thermal inactivation of taro PPO followed first-order kinetics, with an activation energy of E _a?=?422.79?±?0.52?kJ/mol. The textural modification of taro tubers during heating follows the kinetic of the fractional model. It was noticed that the activation energy increased with the concentration of NaCl.     

Alternative Pressing/Ultrafiltration Process for Sugar Beet Valorization: Impact of Pulsed Electric Field and Cossettes Preheating on the Qualitative Characteristics of Juices

Alternative process of sugar beet transformation is investigated by tuning experimental conditions. A three-step process has been set-up: (1) sugar beet cossettes pretreatment by pulsed electric field (PEF) and (or) short preheating to different temperatures; (2) extraction of juice from pre-treated cossettes by pressing; and (3) purification of the expressed juice by ultrafiltration. The PEF treatment was applied to cold (10 °C) and preheated (to 20, 50, 60, 70, and 80 °C) sugar beet cossettes with intensity of E?=?600 V cm^?1 using rectangular monopolar pulses of 100 μs during t _PEF?=?5–20 ms. Experiments were performed with cossettes of three sizes. Control experiments were done without PEF treatment using cold (10 °C) and preheated (to 20–80 °C) cossettes. PEF-treated and (or) preheated cossettes were pressed at 5 bars during 15 min. Afterward, expressed juices obtained from the PEF-treated cossettes at 20 °C and from the untreated ones at 80 °C were purified by dead-end ultrafiltration with stirring (500 rpm) at the temperature of 20 °C by using polyethersulfone membrane with MWCO of 30 kDa. Application of PEF (E?=?600 V cm^?1, t _PEF?=?10 ms, T?=?20 °C) with following pressing of cossettes at 5 bars during 15 min permits to obtain the juice yield Y?=?66,5 %, which is equivalent to that obtained from cossettes preheated to 80 °C and untreated electrically (Y?=?64 %). The energy consumption of cold PEF treatment (≈2–3 Wh/kg) is very attractive as compared to preheating at high temperatures (≈138–194 Wh/kg). Combination of thermal and electrical pretreatments leads to additional softening of sugar beet tissue and to a slightly higher (on 5–10 %) juice yield, but the electroporation of preheated cossettes is more energetically costly. The raw juice expressed from PEF-treated cossettes at 20 °C has higher purity (93.5 %) than juices expressed at 50 °C (92.9 %) and at 80 °C (92.3 %). The temperature increasing from 20 to 80 °C results in a higher juice coloration (5680 IU at 20 °C and 7820 IU at 80 °C) and leads to a higher (on about 35 %) colloids concentration in the expressed juice. The filtrate obtained from the juice expressed at 20 °C with PEF treatment has a higher purity (96 %) than the filtrate obtained from the juice expressed at 80 °C (95.3 %) and its coloration is considerably lower (330 IU versus 1930 IU). In addition, the quantity of proteins and colloids in the filtrate of juice expressed at 20 °C is lower than that in the filtrate of juice expressed at 80 °C     

Characterization of Sultaniye grape (Vitis vinifera L. cv. Sultana) polyphenol oxidase

Polyphenol oxidase (PPO) was extracted from Sultaniye grapes grown in Turkey, and its characteristics in terms of pH and temperature optima, thermal inactivation, kinetic parameters and potency of some PPO inhibitors were studied. Optimum pH and temperature for grape PPO were found to be 3.4 and 30 °C, using catechol as substrate. K_m and V_max values were found to be 44.5 ± 5.47 mm and 0.695 ± 0.0353 OD₄₁₀ min^?1, respectively. Four inhibitors were tested in this study and the most potent inhibitor was sodium metabisulphite, followed by ascorbic acid. From the thermal inactivation studies in the range of 65–80 °C, the half‐life values of the enzyme ranged between 2.6 and 49.5 min. Activation energy (E_a) and Z values were calculated to be 208.5 kJ mol^?1 (r² = 0.9544) and 10.95 °C (r² = 0.9517), respectively.