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.     

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.     

Properties of polyphenol oxidase in mango (Mangifera indica) kernel

Polyphenol oxidase (PPO) activity of filtered extract of ground mango kernel suspension (400 g litre⁻¹) was studied spectrophotometrically at 420 nm using catechol as substrate. The enzyme was most active at pH 6·0 and 25°C. Activity was reduced by 50% at pH values of 5·0 and 7·1, and also at temperatures of 14°C and 30°C. The calculated activation energy and the Michaelis constant (K_m) were 21·4 kcal mol⁻¹ °C⁻¹ and 24·6 mM , respectively. The V_max value was 2·14 units g⁻¹ mango kernel. The time to heat inactivate PPO decreased rapidly to < 10 min with increasing temperature of ⩾ 70°C at 50% activity. © 1998 SCI.     

Partial purification and characterization of polyphenoloxidase from peppermint (Mentha piperita)

Polyphenoloxidase (PPO) of peppermint leaves ( Mentha piperita) was isolated by (NH₄)₂SO₄ precipitation and dialysis. Its pH and temperature optima were 7.0 and 30°C, respectively. On heat-inactivation, half of the activity was lost after 6.5 and 1.5 min of treatment at 70 and 80°C, respectively. Sucrose, (NH₄)₂SO₄, NaCl and KCl appeared to be protective agents of peppermint PPO against thermal denaturation. Km of this enzyme ranged from 6.25×10⁻³ M with catechol to 9.00×10⁻³ M with L-dopa. The I₅₀ values of inhibitors studied on PPO were determined by means of activity percentage (I) diagrams. Values were 1.4×10⁻⁴ M, 1.7×10⁻⁴ M, 9.7×10⁻⁵ M, 2.45×10⁻⁴ M, 2.16×10⁻¹ M, 1.83×10⁻⁵ M, 6.5×10⁻⁵ M, 1.4×10⁻² M, 7.5×10⁻⁵ M, for potassium cyanide, glutathione, ascorbic acid, thiourea, sodium azide, sodium metabisulfite, dithioerythritol, β-mercaptoethanol and sodium diethyl dithiocarbamate respectively. Therefore, sodium metabisulfite was the most effective inhibitor.     

Polyphenoloxidase from DeChaunac grapes

Polyphenoloxidase (PPO) from red grape cultivar, DeChaunac, grown in New York State was isolated and purified 17-fold by using Phenyl Sepharose CL-4B column. Disc gel electrophoresis revealed near homogeneity of three isoenzyme bands. The molecular weight of this enzyme ranged from 73 000 to 85 000. The temperature and pH optima of the purified enzyme were 20 °C and 6.0, respectively. Kinetic studies showed that the thermal inactivation of the PPO followed first-order kinetics, with the activation energy, Ea = 52.39 Kcal mol^?1. The substrate specificity showed a high degree of PPO activity toward o-diphenolic compounds with the highest affinity toward caffeic acid among substrates studied. The apparent K_m values for caffeic acid and 4-methylcatechol as substrates for the Dechaunac PPO were 16 mmol and 25 mmol, respectively. The most potent inhibitors of the PPO were D.L-dithiothreitol and sodium metasulphite at the concentration level of 0.5 mmol.     

A comparative study of polyphenoloxidases from taro (Colocasia esculenta) and potato (Solanum tuberosum var. Romano)

Taro (C. esculenta) is a staple food in many tropical regions. A comparative study of crude polyphenoloxidases from taro (tPPO) and potatoes (pPPO) was carried out to provide information useful for guiding food processing operations. Crude PPO was prepared by cold acetone precipitation using ascorbic acid as antioxidant. The PPO content of taro acetone powder was 770±17 units (mg protein)⁻¹ as compared with 3848±180 units (mg protein)⁻¹ in potato acetone powder. The pH-activity optimum was pH 4.6 for tPPO and pH 6.8 for pPPO. Both enzymes retained >80% activity after incubation at pH 4.5–8 but there was rapid activity loss at pH < 4. The temperature-activity optimum (T_opt) was 30°C for tPPO and 25°C for pPPO with 75 and 27% of their respective maximum activity retained at 60°C. Both tPPO and pPPO were irreversibly inactivated by 10 min heating at 70°C. The activation enthalpy (ΔH^#) and activation entropy (ΔS^#) for tPPO heat-inactivation were 87.4 (±0.1) kJ mol⁻¹ and −56.2 (±4) J mol⁻¹ K⁻¹, respectively. For pPPO, ΔH^# was 59.1 (±0.1) kJ mol⁻¹ whilst ΔS^# was −141 (±4) J mol⁻¹ K⁻¹. The apparent substrate specificity was established from values V_max/K_m as: 4-methylcatechol>chlorogenic acid>dl-dopa>catechol>pyrogallol> dopamine>>caffeic acid for tPPO. There was no detectable activity towards caffeic acid. The substrate specificity for pPPO was: 4-methylcatechol>caffeic acid>pyrogallol>catechol>chlorogenic acid >dl-dopa>dopamine. According to the order of inhibitor effectiveness (sodium metabisulphite>ascorbic acid>NaCl≈ (EDTA), there was a significant lag-phase before increases occurred in the absorbance at 420 nm. Preincubation of PPO with inhibitors increased the extent of inhibition, indicating a direct effect on the structure of the enzyme.     

Biochemical characteristics and thermal inhibition kinetics of polyphenol oxidase extracted from Thompson seedless grape

Polyphenol oxidase (PPO) was isolated from Thompson seedless grape (Vitis vinifera ‘Thompson Seedless’), and its biochemical characteristics were studied. The PPO showed activity to catechol and D, L-DOPA, but not towards monophenol l-Tyrosine, diphenols guaiacol and caffeic acid, and triphenols pyrogallic acid and gallic acid. Apparent Michaelis–Menten constant (K _m) and maximum velocity of the reaction (V _max) values were 45.0 ± 0.05 mM and 500.0 ± 15.3 OD_400 nm/min for catechol, and 34.6 ± 0.03 mM and 384.6 ± 11.7 OD_478 nm/min for D, L-DOPA, respectively. The obtained similar specificity values of V _max/K _m ratio of catechol and D, L-DOPA indicated their similar affinity to Thompson seedless PPO. The most effective inhibitor was l-cysteine, followed in decreasing order by ascorbic acid, sodium metabisulfite, EDTA, NaCl, and citric acid. It was discovered that metal ions of Mg²⁺ and Cu²⁺ increased, while Zn²⁺ and K⁺ reduced the PPO activity. Sugars showed inhibition on the PPO activity, with higher effect by sucrose and lower effect by fructose and glucose. Optimum pH and temperature for grape PPO activity were 6.0 and 25 °C with 10 mM catechol as substrate. The enzyme was heat stable between 10 and 25 °C, but showed significant activity loss at temperatures higher than 40 °C and completely inactivation at 70 °C for 10 min. Thermal inactivation of PPO showed a first-order kinetic with an activation energy (E _a) of 146.1 ± 10.8 kJ/mol at pH 6.0.     

Characterization and inhibition of <Emphasis Type="Italic">Rosmarinus officinalis</Emphasis> L. polyphenoloxidase

Polyphenoloxidase (PPO) from Rosmarinus officinalis L. was fractionated by ammonium sulfate ((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 using 4-methylcatechol, catechol, and pyrogallol as substrates. The protein content of Rosmarinus officinalis L. extracts was determined according to Bradford’s method. Kinetic parameters, K _m and V _max, were calculated from Lineweaver–Burk plots. According to V _max/K _m ratio, 4-methylcatechol was the most suitable substrate. The optimum temperature and pH values were 20, 30 and 30 °C, and 7, 8 and 8 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 partly purified Rosmarinus officinalis L. PPO was spectrophotometrically investigated. For this purpose, ascorbic acid and l-cysteine were used to inhibit the activity of Rosmarinus officinalis L. PPO at different concentrations. From the experimental results, it was found that l-cysteine is a more effective inhibitor than ascorbic acid due to lower K _i values.     

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, K _m and V _max, were calculated from Lineweaver–Burk plots. According to V _max/K _m 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.     

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.     

Effects of temperature on the physicochemical properties of starches extracted from microtubers grown from different potato cultivars

Twelve potato cultivars were grown as microtubers in constant environment chambers at two temperatures in total darkness for 28 days. For starch extracted from microtubers grown at 12 or 22 °C, the amylose content ranged from 25.0 to 29.0% (average 27.3 ± 1.4%) or 29.5 to 32.7% (average 30.9 ± 1.4%), the amylose/amylopectin ratio from 1:2.5 to 1:3.0 (average 1:2.7) or 1:2.1 to 1:2.4 (average 1:2.2) and the phosphorus content from 0.41 to 0.93 g kg⁻¹ (average 0.72 g kg⁻¹) or 0.38 to 1.67 g kg⁻¹ (average 0.97 g kg⁻¹) respectively. Two major fractions (F1 and F2) were obtained for isoamylase‐debranched starch (amylopectin), with the chain length (CL) averaging 56 ± 3 or 55 ± 1 for F1 and 20 ± 1 or 21 ± 1 for F2 from 12 or 22 °C growth temperature respectively. Peak gelatinisation temperature (T_p) and enthalpy (ΔH) were influenced strongly by growth temperature, with T_p ranging from 60.8 to 64.5 °C (average 62.1 ± 1.0 °C) or 66.9 to 69.6 °C (average 68.2 ± 0.9 °C) and ΔH from 13.3 to 16.9 J g⁻¹ (average 14.8 ± 1.0 J g⁻¹) or 11.3 to 19.5 J g⁻¹ (average 16.3 ± 2.4 J g⁻¹) from 12 or 22 °C growth temperature respectively. The collective data generated at 12 °C were generally more comparable to data published elsewhere (but discussed in the text) for the same potato cultivars grown in field plots (Perthshire). However, there were cultivar specific responses to field or constant environment chambers which make direct comparisons between starches produced from the two systems subject to some variation. Copyright © 2004 Society of Chemical Industry     

Characterization of polyphenol oxidase from Uapaca kirkiana fruit

Polyphenol oxidase (PPO), the enzyme responsible for the postharvest spoilage of fruits, was extracted and purified from Uapaca kirkiana peel and pulp by ammonium sulfate precipitation and dialysis. Further purification of peel PPO was carried out by gel filtration chromatography. Optimum pH values were 7 and 8 for peel and pulp PPO, respectively. The optimum temperatures for peel and pulp PPO were 45 and 35 °C, respectively. Inhibition studies of the PPO enzyme were performed using citric acid, sodium azide, sodium metabisulfite and thiourea. The most effective inhibitors were sodium azide and citric acid for both peel and pulp PPO. V_max and K_m values were 13.63 units min^?1 and 4.923 mmol L^?1, respectively, for peel PPO and 14.03 units min^?1 and 5.43 mmol L^?1, respectively, for pulp PPO. Three isoenzymes of Uapaca kirkiana PPO were detected by polyacrylamide gel electrophoresis. One of the isoenzymes could be identified as having a molecular weight of 26 625 Da. Copyright © 2005 Society of Chemical Industry     

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.     

Urea and Heat Unfolding of Cold-Adapted Atlantic Cod(Gadusmorhua)Trypsin and Bovine Trypsin

The reversible unfolding reactions for phenylmethylsulphonyl fluoride (PMSF)-modified trypins from Atlantic cod (cod PMS-trypsin) and cattle (bovine PMS-trypsin) were monitored by fluorescence spectrophotometry as a function of urea concentration and temperature. For urea unfolding at 25°C, the free energy change at zero concentration of urea (ΔG(H₂O)) for cod PMS-trypsin was 11(±4·4) kJ mol⁻¹ compared with 18(±1·14) kJ mol⁻¹ for bovine PMS-trypsin, while the mid-point concentration for urea unfolding curve ([urea]_1/2) was 3·0(±0·57) M and 4·1(±0·16) M, respectively. From studies of enzyme heat unfolding, the mid point temperature of the thermal unfolding curve ( T _m ) was 46(±1·4)°C for cod PMS-trypsin compared with 57(±2)°C for bovine PMS-trypsin. The standard free energy change (Δ G° ) for reversible thermal unfolding of cod PMS-trypsin was 9(±1) kJ mol⁻¹ compared with 19(±1) kJ mol⁻¹ for bovine PMS-trypsin. Values for the enthalpy (Δ H _m ), entropy (Δ S _m ) and heat capacity (Δ C _p ) for heat unfolding are compared. Results from urea and thermal unfolding studies show that cod PMS-trypsin has a significantly lower conformational stability than bovine PMS-trypsin.     

Thermal inactivation of polyphenoloxidase and peroxidase in green coconut (Cocos nucifera) water

Coconut water is an isotonic beverage naturally obtained from the green coconut. After extracted and exposed to air, it is rapidly degraded by enzymes peroxidase (POD) and polyphenoloxidase (PPO). To study the effect of thermal processing on coconut water enzymatic activity, batch process was conducted at three different temperatures, and at eight holding times. The residual activity values suggest the presence of two isoenzymes with different thermal resistances, at least, and a two‐component first‐order model was considered to model the enzymatic inactivation parameters. The decimal reduction time at 86.9 ^°C (D_{86.9 °C}) determined were 6.0 s and 11.3 min for PPO heat labile and heat resistant fractions, respectively, with average z‐value = 5.6 °C (temperature difference required for tenfold change in D). For POD, D_{86.9 °C} = 8.6 s (z = 3.4 °C) for the heat labile fraction was obtained and D_{86.9 °C} = 26.3 min (z = 6.7 °C) for the heat resistant one.     

Some properties of polyphenol oxidase from lily

A study of crude polyphenol oxidase (PPO) from lily bulbs was carried out to provide information useful for guiding food processing operations. Optimum pH for the enzyme activity in the presence of catechol, were 4.0 and 7.0 at room temperature(approximately 20 °C) and the enzyme was stable in the pH range from 5.0 to 6.5 at 4 °C for 10 h. Its optimum temperature was 40 °C and the heat inactivation of the enzyme followed first‐order kinetics. Lily PPO possessed a diphenolase activity toward catechol, catechin and gallic acid; catechin was the best substrate for the enzyme considering the V_max/K_m ratio. The most effective enzyme inhibitor was sodium sulphite, although ascorbic acid, l ‐cysteine and thiourea were also effective inhibitors at high concentration. But NaCl and citric acid were poor inhibitors of the enzyme. Data generated by this study might help to better prevent lily bulbs browning.     

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.     

Effects of hot-water and steam blanching of sliced potato on polyphenol oxidase activity

The worldwide potato production is considered the fourth-most important food sector due to the increasing use of potatoes as raw materials for high-convenience food. Enzymatic browning, due to polyphenol oxidase (PPO), is related to unacceptability by consumer. Among antibrowning agents, thermal treatments are viable alternatives. In this study, the efficacy of hot-water and steam blanching at 80–90 °C of potato slices (1-cm thick) was evaluated in terms of colour changes as well as PPO inactivation kinetics, substrate specificity and transition state parameters. In general, all treatments [1] bleached the slices, [2] inactivated PPO and [3] reduced its kinetic efficiency. Results from thermal inactivation kinetics promoted hot-water blanching at 90 °C for approx. 2 min as the fastest treatment to obtain enzymatic-stable potato slices. Moreover, steam blanching required more energy (53.93 ± 1.24 kJ mol⁻¹) than hot-water treatment (41.41 ± 4.51 kJ mol⁻¹) to reach the transition state and then to unfold the PPO enzyme.     

Physicochemical and microbiological changes in postmortem crayfish (Procambarus clarkii) stored at 4 °C and 25 °C

The physicochemical and microbiological parameters of postmortem crayfish stored at 4 °C and 25 °C were evaluated in order to reduce safety risks of crayfish from temperature abuse during transportation and storage. Results showed that hepatopancreas of postmortem crayfish had significantly higher contents of total volatile basic nitrogen, biogenic amines (BAs) and higher microbial loads than tail meat, regardless of storage temperature. Enterobacteriaceae and Pseudomonas counts reached log 6.41 log₁₀ CFU g⁻¹ and 6.31 log₁₀ CFU g⁻¹ in hepatopancreas of crayfish at 25 ℃ for 6 h. Putrescine and cadaverine were the main BAs in tail meat with levels of 28.18 ± 0.73 mg kg⁻¹ and 187.32 ± 3.57 mg kg⁻¹, respectively, whilst cadaverine, spermidine and spermine were the major BAs in hepatopancreas, reaching 283.45 ± 3.95 mg kg⁻¹, 62.87 ± 9.02 mg kg⁻¹ and 155.31 ± 4.55 mg kg⁻¹, respectively, after postmortem storage at 25 °C for 12 h. With time increasing, Acinetobacter, Flavobacterium, Aeromonas and Chryseobacterium at 25 °C and Acinetobacter, Flavobacterium, Psychrobacter at 4 °C in tail meat as well as Bacteroides and Muribaculaceae at 25 °C, and Acinetobacter, Psychrobacter at 4 °C in hepatopancreas, gradually became the major genus at the end of storage. Based on the results of spoilage microorganisms and biogenic amines, crayfish stored at 4 °C and 25 °C could be edible within 24 h and 6 h, respectively.     

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.