Food and Bioprocess Technology

, Volume 6, Issue 10, pp 2800–2807 | Cite as

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

  • Fuguo Liu
  • Liying Niu
  • Dajing Li
  • Chunquan Liu
  • Bangquan Jin
Original Paper

Abstract

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 Km values for sweet corn POD and waxy corn POD were 11.01 and 23.01 mM, respectively, whereas the H2O2Km 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 (Ea) 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.

Keywords

Peroxidase Sweet corn Waxy corn Kinetic characterization Thermal inactivation 

Introduction

Corn (Zea mays L.), also called maize, is readily available as fresh, canned, or frozen and largely consumed worldwide due to its soft grains, thin shells, and tastefulness (Scott and Eldridge 2005). Sweet corn (Z. mays L. ssp. saccharata Sturt) is a variety of maize with a high sugar content, which is usually picked when immature (milk stage), prepared and eaten as a vegetable, rather than a grain. Waxy corn (Z. mays L. var. ceratina Kulesh), whose endosperm contained only amylopectin and no amylose starch molecule, tastes stickier than other maize varieties. When raw or processed corn is held in frozen storage for extended periods of time, quality deterioration may occur, such as loss of nutrients, undesirable color changes, off-flavor development, and softening of the texture. A high correlation between the quality changes and enzymatic activity was confirmed (Lee and Hammes 1979; Garrote et al. 1987).

Blanching vegetables to inactive endogenous enzymes is a critical step prior to freezing (Barrett et al. 2000; Gonçalves et al. 2007). Among various enzymes, peroxidase (POD) has been the most popular indicator enzyme in the blanching process because of its high concentration in most plant tissues, high thermal stability, and ease of assay (Burnette 1977; Anthon and Barrett 2002; Gonçalves et al. 2010). POD is an oxidoreductase that is widely distributed in nature, whose primary function in plants is the reduction of H2O2 at the expense of oxidation of phenolic compounds (Połata et al. 2009). Normally, POD does not exist as a single enzyme, but as isoforms in plants; hence, its activity is expressed in the form of a number of discrete isoenzymes. Also, POD exists in plant cells in both soluble and membrane-bound forms (Anthon and Barrett 2002), with the former accounting for a higher proportion in most vegetables, suggesting that studies on the thermal stability of POD in blanched and frozen vegetables should focus mainly on the soluble fraction (Morales-Blancas et al. 2002). In particular, some processed products do not require complete inactivation of POD in order to maintain color, texture, flavor, and nutritional quality. Moreover, reduced blanching time would benefit the industry by decreasing energy costs, water use, and clean up costs. The control of blanching to keep POD inactivation to a suitable residual level is important, emphasizing that the need for the analysis of the kinetics of the process in plants has been recommended (Bahçeci et al. 2005).

Because of the high thermostability, the involvement of POD in thermal inactivation in sweet corn has been studied extensively. Yamamoto et al. (1962) investigated heat inactivation over a wide range of temperatures and showed that inactivation of POD in whole kernel corn was biphasic, suggesting the presence of heat-sensitive and heat-resistant fractions. The latter represented 5 % of total enzyme activity and was concentrated in the pericarp. Lee and Hammes (1979) found that POD residual activity in the outer cob and kernels of corn-on-the-cob blanched in steam for variable times had significant correlations with off-flavor development in corn-on-the-cob. Luna et al. (1986) developed a thermokinetic model describing POD inactivation during blanching–cooling of corn-on-the-cob, which allowed the calculation of the POD activity retention for the kernel, outer cob, and central cob. Collins et al. (1996) suggested that the different POD bandings among different sweet genotypes might be involved in their different performance of flavor quality during frozen storage, and Barrett et al. (2000) evaluated POD activity of three corn cultivars which behaved differently as a function of blanching time.

For successful predictions of the residual enzyme activities to minimize the loss of nutritional and sensory properties, it is necessary to know the enzyme distribution and the parameters of the inactivation kinetics; this information is unique for each vegetable, specie, cultivar, and environmental conditions (Agüero et al. 2008). No published information is available in literature on thermal inactivation of POD during blanching of waxy corn, not to mention the differences between sweet corn and waxy corn. Therefore, the objective of the present study was to study the activity and kinetics of POD from two corn cultivars and to examine the thermal inactivation kinetics. The results will help to optimize blanching process for corn, which will certainly be important for corn products development. Non-purified enzyme extract was chosen in order to obtain information relevant to fresh corn processing at the industrial level. The results were also compared with published data for previously studied plant POD.

Materials and Methods

Materials

Sweet corn cobs and waxy corn cobs, at the milk ripe stage as determined by moisture and consistency of the kernel parenchyma, were obtained in a local market in Nanjing, China. Corn samples were stored at 4 °C in husk and all experiments were carried out within 48 h.

Reagents

Hydrogen peroxide (30 %) and guaiacol (99.5 %) were purchased from Sinopharm Chemical Reagent Co. Ltd., China. Sodium phosphate buffer (pH 7.0) was prepared with sodium dihydrogen phosphate and disodium hydrogen phosphate in distilled water obtaining a molar concentration of 0.2 mol/L, the buffer solution was cooled at 4 °C until used. All other chemicals and reagents were of analytical grade. Distilled water was used for all assays.

Extraction and Determination of Crude POD Activity

The intact kernels and germ fraction were separated manually, about 5.0 g samples was homogenized in ice-water bath with 25 mL phosphate buffer (pH 7.0, 4 °C), then centrifuged at 4 °C, 10,000×g for 30 min, the supernatant was collected and divided into 2 mL each sample, and then the samples were quickly cooled in liquid nitrogen for analysis of POD enzyme activity. The POD activity was determined by measuring the increase of absorbance at 470 nm (ε470 = 26.6 mM−1 cm−1) using guaiacol as a phenolic substrate with hydrogen peroxide (Díaz et al. 2001). The reaction mixture contained 0.5 mL of 1.5 % (m/v) guaiacol, 0.2 mL of 1 % (v/v) H2O2, 2.1 mL of 0.1 mol/L phosphate buffer (pH 6.0), and 0.2 mL of the enzyme extract. The blank sample contained the same mixture solution without the enzyme extract. The maximal initial reaction velocity was calculated over 20 to 140 s linear increase in absorbance. One unit of enzymatic activity was defined as amount of enzyme that oxidizes 1 mM of guaiacol per minute at 25 °C and pH 6.0 under assay conditions.

Effect of pH on Enzyme (Extracted from Corn Kernels) Activity

To determine the pH optimum for POD, activity measurements were carried out over a pH range of 3.0 to 9.0. The three buffers used were citrate phosphate (pH 3.0–5.5), sodium phosphate (pH 6.0–7.5), and borate (pH 8.0–9.0). POD activity was measured according to the method described above and expressed as a percentage of the maximum activity.

Enzymatic Kinetics

In order to determine the kinetic properties, POD (extracted from corn kernels) activity was evaluated toward guaiacol concentrations ranging from 1.0 to 20.0 mM and H2O2 concentrations ranging from 0.5 to 10.0 mM. The Michaelis–Menten constants (Km) were determined from Lineweaver–Burk plots at optimum pH conditions.

Thermal Inactivation Experiments

The thermal inactivation of crude POD extracts from sweet corn kernels and waxy corn kernels was studied at constant temperature between 60 and 85 °C at atmospheric pressure. Aliquots of the homogenates were transferred to centrifuge tubes, each tube with 1.0 mL of enzyme extracts. Samples were heated in a circulating water bath to the indicated temperatures for the times specified. The temperature of the water bath was controlled to 0.1 °C. Following heating, the samples were cooled in ice-water to stop thermal inactivation instantaneously and stored on ice until assay.

Data Analysis

All determinations were conducted three times at least. Regression analysis was made by using Microcal Origin 8.5 (Microcal Software, Inc., Northampton, USA). One-way analysis of variance was determined using the Tukey–Krammer test. Differences between means were considered to be significantly different at P < 0.05.

Results and Discussion

POD Distribution in Fresh Corns

POD isoenzyme pattern varied in different corn tissues and within each tissue from different corn varieties (Hamill and Brewbaker 1969). As shown in Table 1, POD activity in the germ fraction was 3.55 times lower than the whole kernel in waxy corn, while there was no significant difference (P > 0.05) between the two parts of sweet corn. Additionally, noticeable variations of POD activity can be observed depending on corn cultivars. Previous work on POD distribution in corn reported that POD activity was most concentrated in the aleurone layer, germ, and pericarp of corn (Gardner et al. 1969). Yamamoto et al. (1962) and Chenchin and Yamamoto (1973) found that after minutes of heating, the activity remaining in the pericarp fractions was much higher than in other tissue fractions.
Table 1

POD activity in sweet corn and waxy corn

Cultivar

Extraction

POD activity (U)

Waxy corn

Intact kernels

11.89 ± 0.14a

Germ fraction

3.36 ± 0.23

Sweet corn

Intact kernels

5.59 ± 0.37

Germ fraction

7.12 ± 0.56

aMean (n = 3) ± standard deviation

Effect of pH on POD Activity

The optimum pH is a key factor in the expression of POD activity, which depends on the hydrogen donor and buffer solutions used in the activity assay. Using guaiacol as the hydrogen donor, and using citrate phosphate, sodium phosphate, and borate as buffer solutions, the assay of POD activity was carried out. It was found that sweet corn and waxy corn performed similar pH profiles of POD activity with lower activity below pH 4.8 or above pH 6.0, and sweet corn showed maximal activity at pH 4.8 while waxy corn at pH 6.0 (Fig. 1). Both enzymes significantly reduced their activities when pH was below 4.0 or above 6.5 with amplitude greater than 40 %. This result was similar to the report by Boyes et al. (1997), which showed that the maximal pH of soluble and membrane-bound POD was 4.9 to 5.1, respectively. With guaiacol as substrate, acidic pH optima have often been reported for POD of corn plants. Mika and Lüthje (2003) isolated two POD isoenzymes from corn root plasma membranes and their highest activities were observed within pH 4.5–5.5 and 5.5–6.0, respectively. Gray and Montgomery (2003) reported that corn steep water POD had a pH optimum of 4.6 on guaiacol, but atypically, the activity was within 90 % of the maximum over a broad pH range (pH 3.7–5.2). In general, POD from plant showed maximum activity at or near neutral pH although other studies used ABTS (Duarte-Vázquez et al. 2007) or phenol (Motamed et al. 2009) as hydrogen donor.
Fig. 1

Fresh corn POD activity (relative activity to maximal value) as a function of pH (vertical bars represent standard deviations)

Kinetic Studies

Generally, POD specifically use H2O2 as substrate but can employ a number of hydrogen donors such as guaiacol (Duarte-Vázquez et al. 2001), which may prevent the active site of POD from being inactivated by a high concentration of H2O2. Hence, the experiments were carried out at varying concentrations of guaiacol and H2O2. The apparent Km values of guaiacol and H2O2 were then calculated using the Lineweaver–Burk reciprocal plot graphic method. Figure 2a, b shows variations in initial velocity versus guaiacol concentration for sweet corn and waxy corn, respectively, and the catalytic velocity was enhanced with increasing guaiacol concentration. Obviously, the curves showed Michaelis–Menten kinetics but with different values of kinetic parameters between the two cultivars: Km for guaiacol was found to be 11.01 mM for sweet corn and 23.01 mM for waxy corn (Fig. 2a, b, insert), indicating that waxy corn possessed a greater tolerance for guaiacol than sweet corn. The Km values of guaiacol appeared to be higher than those reported for the POD from green peas (Halpin et al. 1989), turnip roots (Duarte-Vázquez et al. 2001), carrot (Soysal and Söylemez 2005), and wheat grass (Lai et al. 2006), but much lower than roots of red beet (Rudrappa et al. 2007).
Fig. 2

Substrate saturation curves and Lineweaver–Burk plots (insets) of POD activity in presence of different concentrations of a guaiacol (for sweet corn); b guaiacol (for waxy corn); and with c H2O2 (for sweet corn); d H2O2 (for waxy corn) as fixed substrate (vertical bars represent standard deviations)

In addition, a similar approach was then taken to determine the kinetic parameters for H2O2 for both corns at 20 mM guaiacol. As shown in Fig. 2c, d, when the H2O2 concentration was increased, the activity increased to reach saturation at 3.5 mM for sweet corn and 5.0 mM for waxy corn, and then decreased with H2O2 concentration above the saturation. Hence, these did not follow pure Michaelis–Menten kinetics. The forms of these graphs were typical of enzymes exhibiting substrate inhibition. At the same time, it was found that the Km values for H2O2 calculated by Lineweaver–Burk reciprocal plots was identical to that estimated from the substrate saturation curves, Km values were 2.85 and 0.33 mM for sweet corn and waxy corn, respectively (Fig. 2c, d, inset). Noticeably, the Km value of waxy corn was eight times lower than sweet corn, which implied that waxy corn POD had a relatively high affinity for H2O2. Other reported Km values for H2O2 by using guaiacol include 7.2 mM for green pea (Halpin et al. 1989), 1.4 mM for carrot (Soysal and Söylemez 2005), and 50.68 and 18.18 mM for two different strawberry cultivars (Chisari et al. 2007), but no inhibition by substrate concentration was observed between them.

Thermal Inactivation Kinetics of POD Crude Extracts

The remaining activities of POD from sweet corn and waxy corn were studied as a function of temperature to determine their thermodynamic properties. As expected, the extent of POD denaturation increased with increasing temperature and treatment time (Fig. 3). POD activities were reduced by approximately 90 % at 70 °C after 3 and 5 min of heat treatment, for sweet corn and waxy corn, respectively. This behavior compared well with the report by Yemenicioğlu et al. (1998), who reported that the soluble POD from fresh pinto beans lost almost 97 % of its activity after being heated for 5 min at 70 °C. The semi-log plots of the residual activity of POD from sweet corn (Fig. 3a) and POD from waxy corn (Fig. 3b) versus heating time were linear at all temperatures studied (R2 > 0.860), which was consistent with inactivation by means of a simple first-order (monophasic) process for both samples. Monophasic behavior of the enzyme inactivation at high temperature could be due to the rapid inactivation of the heat-labile fraction of the enzyme during the first seconds of treatment (Ganjloo et al. 2011). This study was in good agreement with the results of Anthon et al. and Serrano-Martínez et al. (2002–2011), who reported a simple first-order inactivation of POD from carrots and potatoes (Anthon and Barrett 2002), tomato juice (Anthon et al. 2002), red pepper (Serrano-Martínez et al. 2008), table grape (Fortea et al. 2009), and red alga (Fortea et al. 2011). However, some researchers observed biphasic curves during thermal inactivation of POD in vegetables (Yemenicioğlu et al. 1998a, b; Morales-Blancas et al. 2002; Garrote et al. 2004; Cruz et al. 2006; Agüero et al. 2008). Rudra et al. (2008) also found that two-fraction first-order model was the best model to describe the POD inactivation kinetics in coriander leaves (R2 > 0.97). Moreover, Soysal and Söylemez (2005) and Agüero et al. (2008) found that inactivation kinetics of POD showed a biphasic first-order model below 70 °C, while above 70 °C, POD showed monophasic first-order behavior.
Fig. 3

POD inactivation curves at different temperature for a sweet corn and b waxy corn as represented by first-order kinetic equation (standard deviation bars are smaller than the symbol size)

From the slopes of these lines, the inactivation rate constants (k) were calculated by linear regression according to the equation (Anthon and Barrett 2002):
$$ \log \left( {{A_t \left/ {A_0 } \right.}} \right)=-\left( {{k \left/ {2.303 } \right.}} \right)t $$
where A0 is the initial activity, and At is the remaining activity at time t. The rate constant increased with the heating temperature, indicating that POD from both sweet corn and waxy corn was less thermostable at higher temperature, and POD from waxy corn was more heat-resistant as shown by its smaller inactivation rate constants (Table 2). Activation energy (Ea) value is an important parameter for the inactivation of enzymes. Arrhenius plots (Fig. 4) of the constants in Table 1 were also linear and showed little difference between the two corn cultivars. This linearity indicated that the inactivation in fresh corn POD occurred through a unique temperature-dependent mechanism, such as protein unfolding (Fortea et al. 2011). From the slopes of these lines, Ea of 114.36 ± 6.97 kJ/mol for sweet corn POD and 119.72 ± 5.11 kJ/mol for waxy POD were calculated by the equation (Anthon and Barrett 2002) below:
Table 2

Thermal inactivation parameters for POD in sweet corn and waxy corn

Sample

T (°C)

k (min−1)

D (min)

R2

Ea (kJ/mol)

ZT (°C)

Sweet corn

60

0.18 ± 0.02a

12.76 ± 1.43

0.958

 

65

0.37 ± 0.01

6.22 ± 0.20

0.961

70

0.77 ± 0.03

2.98 ± 0.12

0.972

75

1.27 ± 0.20

1.82 ± 0.27

0.988

85

3.26 ± 0.25

0.71 ± 0.06

0.926

 

114.36 ± 6.97

20.08 ± 0.09

Waxy corn

60

0.10 ± 0.01

24.27 ± 1.58

0.860

 

65

0.26 ± 0.03

8.91 ± 0.93

0.925

70

0.54 ± 0.07

4.27 ± 0.53

0.946

75

0.88 ± 0.13

2.61 ± 0.51

0.991

80

1.18 ± 0.13

1.96 ± 0.26

0.948

 

119.72 ± 5.11

18.38 ± 0.44

aMean (n = 3) ± standard deviation

Fig. 4

Arrhenius plot showing the effect of temperature on the rate constant for the thermal inactivation of POD crude extract from sweet corn and waxy corn (standard deviation bars are smaller than the symbol size)

$$ \ln (k)=-{{{{E_{\mathrm{a}}}}} \left/ {RT+c } \right.} $$
where R is the gas constant (8.314 J mol−1 K−1) and T is the temperature in Kelvin. The result proved that sweet corn POD had a greater sensitivity to temperature change than that of waxy corn POD. The Ea values observed for POD from both sweet corn and waxy corn were higher than other POD reported, such as corn-on-the-cob (30.96 ± 1.67 kJ/mol, Lee and Hammes 1979; 74.89–87.86 kJ/mol, Naveh et al. 1982; 76.15–82.84 kJ/mol, Luna et al. 1986), pumpkin (86.20 ± 5.57 kJ/mol, Gonçalves et al. 2007), and seedless guava (96.39 ± 4 kJ/mol, Ganjloo et al. 2011); similar to that obtained for POD from siam weed (120.14 kJ/mol, Rani and Abraham 2006) and red alga (121.6 kJ/mol, Fortea et al. 2011); and lower than those reported for tomato (149 kJ/mol, Ercan and Soysal 2011), carrot (151.40 ± 5.44 kJ/mol, Gonçalves et al. 2010; 480 kJ/mol, Anthon and Barrett 2002), red pepper (151 kJ/mol, Serrano-Martínez et al. 2008), table grape (271.9 kJ/mol, Fortea et al. 2009), and potatoes (478 kJ/mol, Anthon and Barrett 2002). Such differences may be attributed to differences in plant sources of the enzyme, degree of ripening, or experimental methodology.
To further study the thermostability, the more commonly used D and ZT values for the enzyme systems were also estimated. D value is the time required to reduce the enzyme activity to 10 % of its original value, which is calculated from k according to their relation (Anthon and Barrett 2002): D = 2.303/k. As summarized in Table 2, D values for both corns decreased with increasing temperature, indicating a faster inactivation at higher temperature. At the same time, D values of POD from sweet corn were much lower than that from waxy corn at a constant temperature, implying again that POD from sweet corn was less stable than that from waxy corn. The effect of temperature on D values is shown in Fig. 5, where the slope of the curves represents −1/ZT. Table 2 showed the calculated ZT values for POD from sweet corn and waxy corn, between which there were only slight differences. However, Naveh et al. (1982) reported significantly higher ZT values up to 39.3 °C, for the inactivation of POD from corn-on-the-cob, which was probably due to different sources and different experimental conditions. In general, low ZT values are thought to indicate greater sensitivity to heat.
Fig. 5

Variation of decimal reduction times with temperature for POD crude extract from sweet corn and waxy corn (vertical bars represent standard deviations)

Conclusion

The present results indicated that POD activity in aqueous extracts from sweet corn and waxy corn differed significantly in total kernel or germ fraction and varied by pH and substrate concentrations. The enzyme kinetics followed Michaelis–Menten equation, showing different values of POD kinetics parameters between the two cultivars. In addition, the thermostability of POD is very important due to its negative effect on the color and flavor of corn during storage. POD from sweet corn and waxy corn showed diverse heat sensitivities with variations in K, Ea, D, and ZT. Taking those values into consideration, POD from waxy corn was found to be more heat stable than sweet corn, which implies that the blanching parameters should be optimized discriminately.

Notes

Acknowledgments

The authors would like to thank the financial support from Jiangsu Academy of Agricultural Sciences for Innovation in Agriculture Science and Technology [project no. CX (11) 4026].

References

  1. Agüero, M. V., Ansorena, M. R., Roura, S. I., & del Valle, C. E. (2008). Thermal inactivation of peroxidase during blanching of butternut squash. LWT-Food Science and Technology, 41(3), 401–407.CrossRefGoogle Scholar
  2. Anthon, G. E., & Barrett, D. M. (2002). Kinetic parameters for the thermal inactivation of quality-related enzymes in carrots and potatoes. Journal of Agricultural and Food Chemistry, 50(14), 4119–4125.CrossRefGoogle Scholar
  3. Anthon, G. E., Sekine, Y., Watanabe, N., & Barrett, D. M. (2002). Thermal inactivation of pectin methylesterase, polygalacturonase, and peroxidase in tomato juice. Journal of Agricultural and Food Chemistry, 50(21), 6153–6159.CrossRefGoogle Scholar
  4. Bahçeci, K. S., Serpen, A., Gökmen, V., & Acar, J. (2005). Study of lipoxigenase and peroxidase as indicator enzymes in green beans: change of enzyme activity, ascorbic acid and chlorophylls during frozen storage. Journal of Food Engineering, 66(2), 187–192.CrossRefGoogle Scholar
  5. Barrett, D. M., Garcia, E. L., Russell, G. F., Ramirez, E., & Shirazi, A. (2000). Blanch time and cultivar effects on quality of frozen and stored corn and broccoli. Journal of Food Science, 65(3), 534–540.CrossRefGoogle Scholar
  6. Boyes, S., Chevis, P., & Perera, C. (1997). Peroxidase isoforms of corn kernels and corn on the cob: preparation and characteristics. LWT-Food Science and Technology, 30(2), 192–201.CrossRefGoogle Scholar
  7. Burnette, F. S. (1977). Peroxidase and its relationship to food flavor and quality a review. Journal of Food Science, 42(1), 1–6.CrossRefGoogle Scholar
  8. Chenchin, E. E., & Yamamoto, H. Y. (1973). Distribution and heat inactivation of peroxidase isoenzymes in sweet corn. Journal of Food Science, 38(1), 40–42.CrossRefGoogle Scholar
  9. Chisari, M., Barbagallo, R. N., & Spagna, G. (2007). Characterization of polyphenol oxidase and peroxidase and influence on browning of cold stored strawberry fruit. Journal of Agricultural and Food Chemistry, 55(9), 3469–3476.CrossRefGoogle Scholar
  10. Collins, J. K., Biles, C. L., Wann, E. V., Perkins-Veazie, P., & Maness, N. (1996). Flavour qualities of frozen sweetcorn are affected by genotype and blanching. Journal of the Science of Food and Agriculture, 722(4), 425–429.CrossRefGoogle Scholar
  11. Cruz, R. M. S., Vieira, M. C., & Silva, C. L. M. (2006). Effect of heat and thermosonication treatments on peroxidase inactivation kinetics in watercress (Nasturtium officinale). Journal of Food Engineering, 72(1), 8–15.CrossRefGoogle Scholar
  12. Díaz, J., Bernal, A., Pomar, F., & Marino, F. (2001). Induction of shikimate dehydrogenase and peroxidase in pepper (Capsicum annuum L.) seedings in response to copper stress and its relation to linification. Plant Sciences, 16(1), 179–188.CrossRefGoogle Scholar
  13. Duarte-Vázquez, M. A., García-Almendárez, B. E., Regalado, C., & Whitaker, J. R. (2001). Purification and properties of a neutral peroxidase isozyme from turnip (Brassica napus L. var. purple top white globe) roots. Journal of Agricultural and Food Chemistry, 49(9), 4450–4455.CrossRefGoogle Scholar
  14. Duarte-Vázquez, M. A., García-Padilla, S., García-Almendárez, B. E., Whitaker, J. R., & Regalado, C. (2007). Broccoli processing wastes as a source of peroxidase. Journal of Agricultural and Food Chemistry, 55(25), 10396–10404.CrossRefGoogle Scholar
  15. Ercan, S. S., & Soysal, Ç. (2011). Effect of ultrasound and temperature on tomato peroxidase. Ultrasonics Sonochemistry, 18(2), 689–695.CrossRefGoogle Scholar
  16. Fortea, M. I., López-Miranda, S., Serrano-Martínez, A., Carreño, J., & Núñez-Delicado, E. (2009). Kinetic characterisation and thermal inactivation study of polyphenol oxidase and peroxidase from table grape (Crimson Seedless). Food Chemistry, 113(4), 1008–1014.CrossRefGoogle Scholar
  17. Fortea, M. I., López-Miranda, S., Serrano-Martínez, A., Hernández-Sánchez, P., Zafrilla, M. P., Martínez-Cachá, A., et al. (2011). Kinetic characterisation and thermal inactivation study of red alga (Mastocarpus stellatus) peroxidase. Food Chemistry, 127(1), 1091–1096.CrossRefGoogle Scholar
  18. Ganjloo, A., Rahman, R. A., Osman, A., Bakar, J., & Bimakr, M. (2011). Kinetics of crude peroxidase inactivation and color changes of thermally treated seedless guava (Psidium guajava L.). Food and Bioprocess Technology, 4(8), 1442–1449.CrossRefGoogle Scholar
  19. Gardner, H. W., Inglett, G. E., & Anderson, R. A. (1969). Inactivation of peroxidase as a function of corn processing. Cereal Chemistry, 46(6), 626–634.Google Scholar
  20. Garrote, R. L., Luna, J. A., Silva, E. R., & Bertone, R. A. (1987). Prediction of residual peroxidase activity in the blanching-cooling of corn-on-the-cob and its relation to off-flavor development in frozen storage. Journal of Food Science, 52(1), 232–233.CrossRefGoogle Scholar
  21. Garrote, R. L., Silva, E. R., Bertone, R. A., & Roa, R. D. (2004). Predicting the end point of a blanching process. LWT-Food Science and Technology, 37(3), 309–315.CrossRefGoogle Scholar
  22. Gonçalves, E. M., Pinheiro, J., Abreu, M., Brandão, T. R. S., & Silva, C. L. M. (2007). Modelling the kinetics of peroxidase inactivation, colour and texture changes of pumpkin (Cucurbita maxima L.) during blanching. Journal of Food Engineering, 81(4), 693–701.CrossRefGoogle Scholar
  23. Gonçalves, E. M., Pinheiro, J., Abreu, M., Brandão, T. R. S., & Silva, C. L. M. (2010). Carrot (Daucus carota L.) peroxidase inactivation, phenolic content and physical changes kinetics due to blanching. Journal of Food Engineering, 97(4), 574–581.CrossRefGoogle Scholar
  24. Gray, J. S. S., & Montgomery, R. (2003). Purification and characterization of a peroxidase from corn steep water. Journal of Agricultural and Food Chemistry, 51(6), 1592–1601.CrossRefGoogle Scholar
  25. Halpin, B., Pressey, R., Jen, J., & Mondy, N. (1989). Purification and characterization of peroxidase isoenzymes from green peas (Pisum sativum). Journal of Food Science, 54(3), 644–649.CrossRefGoogle Scholar
  26. Hamill, D. E., & Brewbaker, J. L. (1969). Isoenzyme polymorphism in flowering plants. IV. the peroxidase isoenzyme of maize (Zea mays). Physiologia Plantarum, 22(5), 945–958.CrossRefGoogle Scholar
  27. Lai, L., Wang, D., Chang, C., & Wang, C. (2006). Catalytic characteristics of peroxidase from wheat grass. Journal of Agricultural and Food Chemistry, 54(22), 8611–8616.CrossRefGoogle Scholar
  28. Lee, Y. C., & Hammes, J. K. (1979). Heat inactivation of peroxidase in corn-on-the-cob. Journal of Food Science, 44(3), 785–787.CrossRefGoogle Scholar
  29. Luna, J. A., Garrote, R. L., & Bressan, J. A. (1986). Thermo-kinetic modeling of peroxidase inactivation during blanching–cooling of corn on the cob. Journal of Food Science, 51(1), 141–145.CrossRefGoogle Scholar
  30. Mika, A., & Lüthje, S. (2003). Properties of guaiacol peroxidase activities isolated from corn root plasma membranes. Plant Physiology, 132(3), 1489–1498.CrossRefGoogle Scholar
  31. Morales-Blancas, E. F., Chandia, V. E., & Cisneros-Zevallos, L. (2002). Thermal inactivation kinetics of peroxidase and lipoxygenase from broccoli, green asparagus and carrots. Journal of Food Science, 67(1), 146–154.CrossRefGoogle Scholar
  32. Motamed, S., Ghaemmaghami, F., & Alemzadeh, I. (2009). Turnip (Brassica rapa) peroxidase: purification and characterization. Industrial and Engineering Chemistry Research, 48(23), 10614–10618.CrossRefGoogle Scholar
  33. Naveh, D., Mizrahi, S., & Kopelman, I. J. (1982). Kinetics of peroxidase deactivation in blanching of corn on the cob. Journal of Agricultural and Food Chemistry, 30(5), 967–970.CrossRefGoogle Scholar
  34. Połata, H., Wilińska, A., Bryjak, J., & Polakovič, M. (2009). Thermal inactivation kinetics of vegetable peroxidases. Journal of Food Engineering, 91(3), 387–391.CrossRefGoogle Scholar
  35. Rani, D. N., & Abraham, T. E. (2006). Kinetic study of a purified anionic peroxidase isolated from Eupatorium odoratum and its novel application as time temperature indicator for food materials. Journal of Food Engineering, 77(3), 594–600.CrossRefGoogle Scholar
  36. Rudra, S. G., Shivhare, U. S., Basu, S., & Sarkar, B. C. (2008). Thermal inactivation kinetics of peroxidase in coriander leaves. Food and Bioprocess Technology, 1(2), 187–195.CrossRefGoogle Scholar
  37. Rudrappa, T., Lakshmanan, V., Kaunain, R., Singara, N. M., & Neelwarne, B. (2007). Purification and characterization of an intracellular peroxidase from genetically transformed roots of red beet (Beta vulgaris L.). Food Chemistry, 105(3), 1312–1320.CrossRefGoogle Scholar
  38. Scott, C. E., & Eldridge, A. L. (2005). Comparison of carotenoid content in fresh, frozen and canned corn. Journal of Food Composition and Analysis, 18(6), 551–559.CrossRefGoogle Scholar
  39. Serrano-Martínez, A., Fortea, M. I., del Amor, F. M., & Núñez-Delicado, E. (2008). Kinetic characterisation and thermal inactivation study of partially purified red pepper (Capsicum annuum L.) peroxidase. Food Chemistry, 107(1), 193–199.CrossRefGoogle Scholar
  40. Soysal, Ç., & Söylemez, Z. (2005). Kinetics and inactivation of carrot peroxidase by heat treatment. Journal of Food Engineering, 68(3), 349–356.CrossRefGoogle Scholar
  41. Yamamoto, H. Y., Steinberg, M. P., & Nelson, A. I. (1962). Kinetic studies on the heat inactivation of peroxidase in sweet corn. Journal of Food Science, 27(2), 113–119.CrossRefGoogle Scholar
  42. Yemenicioğlu, A., Özkan, M., Velioğlu, S., & Cemeroğlu, B. (1998). Thermal inactivation kinetics of peroxidase and lipoxygenase from fresh pinto beans (Phaseolus vulgaris). Zeitschrift für Lebensmittel-Untersuchung und-Forschung A, 206(4), 294–296.CrossRefGoogle Scholar
  43. Yemenicioğlu, A., Özkan, M., & Cemeroglu, B. (1998a). Thermal stabilities of peroxidases from fresh pinto beans. Journal of Food Science, 63(6), 987–990.CrossRefGoogle Scholar
  44. Yemenicioğlu, A., Özkan, M., & Cemeroglu, B. (1998b). Partial purification and thermal characterization of peroxidase from okra (Hibiscus esculentum). Journal of Agricultural and Food Chemistry, 46(10), 4158–4163.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Fuguo Liu
    • 1
    • 2
    • 3
  • Liying Niu
    • 1
    • 2
  • Dajing Li
    • 1
    • 2
  • Chunquan Liu
    • 1
    • 2
  • Bangquan Jin
    • 3
  1. 1.Institute of Farm Product ProcessingJiangsu Academy of Agricultural SciencesNanjingPeople’s Republic of China
  2. 2.Engineering Research Center for Agricultural Products ProcessingNational Agricultural Science and Technology Innovation Center in East ChinaNanjingPeople’s Republic of China
  3. 3.Department of Food Science and Technology, Ginling CollegeNanjing Normal UniversityNanjingPeople’s Republic of China

Personalised recommendations