Extraction and characterisation of pectin methylesterase from black carrot (Daucus carota L.)

This study was carried out to determine some of the biochemical properties of pectin methylesterase (PME) from black carrot. The enzyme showed very high activity in a broad pH range of 6.5–8.5, with the optimum pH occurring at 7.5. The optimum temperature for maximal PME activity was found to be 55 °C. NaCl enhanced PME activity, particularly at 0.2 M. K_m and V_max values for black carrot PME using apple pectin as substrate were found to be 2.14 mg/ml (r² = 0.988) and 3.75 units/ml, respectively. The enzyme was stable between the temperatures of 30–50 °C/5 min whereas it lost nearly all of its activity at 70 °C/5 min. E_a and Z values were found to be 196.8 kJmol⁻¹ (r² = 0.996) and 2.16 °C (r² = 0.995), respectively.     

Effect of preheating on potato texture

Preheating potatoes at 50 to 80°C has a firming effect on the cooked potato tissue. This effect is particularly pronounced at a preheating temperature of 60 to 70°C followed by cooling. Several theories have been presented in the literature to explain this firming effect: retrogradation of starch, leaching of amylose, stabilization of the middle lamellae and cell walls by the activation of the pectin methylesterase (PME) enzyme, and by the release of calcium from gelatinized starch and the formation of calcium bridges between pectin molecules. Most probably, none of these theories alone can explain the phenomenon and more than one mechanism seems to be involved. Some of these mechanisms seem to be interdependent. As an example, calcium could be considered as a link all the way through release after starch gelatinization to cross‐linking pectin substances in the cell wall and the middle lamellae, which has been demethylated by the PME enzyme. More research and “clear cut” experiments are needed in order to elucidate the role of each mechanism, especially which of them is the main contributor to the process of firming. Most probably, the calcium‐pectin‐PME mechanism plays a secondary role, that is, it only retards the collapse of the tissue structure that would otherwise occur during the final heating without preheating, and it is not the main factor of firmness.     

Pectin methylesterase catalyzed firming effects on low temperature blanched vegetables

The effect of low temperature blanching on firmness of eight vegetables was studied. Preheating vegetables at low temperatures prior to a conventional blanch resulted in firmer products. Temperature and time had significant effects on texture, with temperature the most influential. Under optimal conditions, firmness improvements in preheated vegetables as compared to blanched controls were: Bok choy—3.0 × (65 °C, 45 min); Chinese cabbage—1.8 × (55 °C, 45 min); cabbage—1.6 × (65 °C, 15 min); green bell peppers—1.36 × (70 °C, 15 min); sugar snap peas—1.7 × (65 °C, 30 min); carrots—2.1 × (60 °C, 15 min) and broccoli—2.9 × (60 °C, 15 min). Thermal stability and optimal temperature for pectin methylesterase in homogenates from these vegetables were also analyzed. The relationship between optimum preheating conditions for textural integrity and pectin methylesterase activity is discussed.     

The effect of low-temperature blanching on the quality of fresh and frozen/thawed mashed potatoes

The effect of low‐temperature blanching (LTB) prior to cooking on colour, textural, firmness and oscillatory parameters, sensory attributes and overall acceptability of either fresh or frozen/thawed mashed potatoes was studied using response surface methodology (RSM) to establish the optimum temperature and time for blanching in both types of mashed potatoes. A central composite rotatable design was used to study the effects of variation in levels of blanching temperature (57.93–72.07 °C) and time (15.86–44.14 min) on the quality parameters. Stationary points showing maximum thickening had critical temperatures (approximately 67–69 °C) and times (approximately 26–30 min) in the ranges of temperature and time used for each independent variable for both fresh and frozen/thawed mashed potato. Results showed a high correlation between structural reinforcement and overall acceptability under optimum experimental blanching conditions. This demonstrates the potential of this experimental approach in terms of tailoring physical properties to predetermined levels in order to meet consumer preferences in mashed potatoes, and of altering the changes that occur after freezing and thawing.     

Towards a better understanding of the pectin structure-function relationship in broccoli during processing: Part I—macroscopic and molecular analyses

To investigate the structure-function relationship of pectin during (pre)processing, broccoli samples (Brassica oleracea L. cultivar italica) were subjected to one of the following pretreatments: (i) low-temperature blanching (LTB), (ii) LTB in combination with Ca²⁺ infusion, (iii) high-pressure pretreatment (HP), (iv) HP in combination with Ca²⁺ infusion, or (v) no pretreatment (control sample), whether or not in combination with a thermal treatment of 15 min at 90 °C. The macroscopic attributes of broccoli were linked to the chemical structure of broccoli pectin. By enhancing the cross-linking of pectic polymers, both LTB and HP reduced the texture loss that occurred during thermal processing of broccoli. During these pretreatments, homogalacturonan was de-esterified by pectin methylesterase, which led to changes in pectin solubility. When LTB or HP was combined with Ca²⁺ infusion, changes in the structure of pectin occurred, however not always reflected at the macroscopic level. The degree of esterification of pectin in Ca²⁺-soaked broccoli samples was lower compared to non-Ca²⁺-soaked samples and, in addition, a higher amount of ionically cross-linked pectin was retrieved.     

Mild-Heat and High-Pressure Inactivation of Carrot Pectin Methylesterase: A Kinetic Study

ABSTRACT: Pectin methylesterase (PME) was extracted from carrots and purified by affinity chromatography. The thermal high-pressure inactivation of the PME was investigated in a model system. Under these conditions, the (thermo) stable fraction is not inactivated and the isobaric-isothermal inactivation followed a fractional-conversion model. At lower pressures (< 300 MPa) and higher temperatures (> 50°C), an antagonistic effect of pressure and heat was observed. A 2nd- and 3rd-degree polynomial model (derived from available thermodynamic model) was successfully used to describe the heat pressure dependence of the inactivation rate constants. From the purified carrot PME sample, the thermostable PME fraction was isolated. The thermal inactivation of this fraction followed first-order kinetics.     

ACEROLA's CLONES OF INDUSTRIAL INTEREST

In order to know which clone of acerola is better for acerola industrialization, we studied the pectin methylesterase (PME) specific activity, pectin content and vitamin C content in five different clones of acerola. The pectin yield varied from 1.37 to 2.99% and the highest content of pectin occurred in clones 3 and 5. Ascorbic acid varied significantly from 1157.5 to 1735.5 mg/100 g of pulp in the five clones. The highest content of vitamin C occurred in clone 4. The PME specific activity varied from 0.79 to 2.92 units g ^?1 /g of pulp and the highest values occurred in clone 2. We also studied the optimum temperature and the optimum pH of this enzyme. Clones 1, 2, 4 and 5 showed optimum temperature at 90C. Clone 3 showed practically the same specific activity at all temperatures studied. Clones 1 and 4 showed an optimum pH of 9.0 and clone numbers 2, 3 and 5 showed a pH optimum at 8.5.     

High-pressure treatment of cloudy apple juice

A factorial design with four factors (pressure, holding time, temperature and waiting period between crushing of apple and high-pressure (HP) treatment) was built up to study the effect of HP treatment on pectin methylesterase activity (PME, EC 3.1.1.11) in cloudy apple juice. PME activity was measured by the methanol release during the storage of samples at 4 °C for 1 month. Only the holding time of the treatment and three interactions between factors (holding time×temperature, pressure×waiting and holding time×waiting) had significant effects on the PME level. Increasing the pressure or the holding time at moderate temperatures (15-40 °C) increased the activity. PME purified from apple was pressure stable in the range 100-600 MPa at 25 °C. The soluble pectin content was not changed after HP treatment. The particles size increased with the temperature reached during the treatment, in agreement with a diffusion-limited-aggregation model but were independent of pressure and holding time. In a separate experiment we found a positive correlation between the PME activity and the residual content of the catechins suggesting that polyphenoloxidase (PPO, EC 1.14.18.1) and oxidized polyphenols play a role in the PME activity level after HP treatment.