Effects of Carbon Dioxide in High-Pressure Processing on Pectinmethylesterase in Single-strength Orange Juice

ABSTRACT: High-pressure processing (HPP) in combination with added CO was examined for efficacy in inactivating Valencia orange juice pectinmethylesterase (PME). Noncarbonated juice and carbonated juice were subjected to a range of conditions from 200 to 600 MPa, 30 to 300 s dwell time at pressure, and 15 to 50 °C at final processing temperature. Processing pressure magnitude, temperature, and dwell time were significant factors (p < 0.001) in the inactivation of PME. At 600 MPa and 25 °C final processing temperature, the addition of 12 psig carbon dioxide resulted in a decrease of pressure processing time from 346 s to 111 s, while achieving an equivalent reduction in PME activity.     

Selection of Process Conditions for High Pressure Pasteurization of Sea Buckthorn Juice Retaining High Antioxidant Activity

Sea buckthorn berries juice is a nutritious beverage, rich in vitamin C and carotenoids with high antioxidant activity. The main requirements for a freshly squeezed sea buckthorn juice production are the cloud stability and antioxidant activity retention after processing. Appropriate process technologies and conditions have to be applied in order to inactivate pectin methyl esterase (PME), responsible for cloud loss, while maintaining the nutritional characteristics and antioxidant activity of the juice. The objectives of the present work were to study and model the effect of thermal treatment and high pressure (HP) processing on the inactivation kinetics of endogenous PME and on total antioxidant activity alteration. Thermal treatment significantly affected PME inactivation and residual antioxidant activity. Processing even at mild process conditions (60 °C for 1 min) resulted in 2.5-fold antioxidant activity reduction and 50 % PME inactivation compared to untreated sample. Pressure and temperature acted synergistically for PME inactivation that followed first-order kinetics with a residual PME activity at all pressure–temperature combinations used (200–600 MPa and 25–35 °C). The effect of temperature and pressure on the inactivation rate constants was expressed through the activation energy and activation volume, respectively. Values of 163 kJ/mol and ?17 mL/mol at reference pressure of 600 MPa and reference temperature of 35 °C were estimated, respectively. Antioxidant activity of the samples was expressed through the determination of the effective concentration (EC₅₀). A slight increase in sea buckthorn antioxidant activity when applying pressures (200–600 MPa) at ambient temperature (25 °C) was observed compared to the corresponding value of untreated juice. Processing at higher temperatures did not significantly alter the total antioxidant activity of sea buckthorn juice. For sample treated at 600 MPa–35 °C for 5 min, a 5 % reduction of total antioxidant activity was observed. These conditions are proposed as effective process conditions for sea buckthorn juice cold pasteurization, based on the higher antioxidant activity retention and simultaneous PME inactivation.     

Comparison of Microbial Inactivation and Rheological Characteristics of Mango Pulp after High Hydrostatic Pressure Treatment and High Temperature Short Time Treatment

The effects of high hydrostatic pressure (HHP) treatments at pressures of 300–600 MPa for 1–20 min and of high-temperature, short-time (HTST) treatment on the inactivation of natural microorganisms in blanched mango pulp (BMP) and unblanched mango pulp (UBMP) were investigated. No yeasts, molds, or aerobic bacteria were detected in BMP or UBMP after HHP treatments at 300 MPa/15 min, 400 MPa/5 min, 500 MPa/2.5 min, and 600 MPa/1 min and HTST treatment at 110 °C/8.6 s. Therefore, these conditions were selected to study the effects of HHP and HTST treatments on pectin methylesterase (PME) activity, water-soluble pectin (WSP) levels, and the rheological characteristics of UBMP and BMP. HHP treatment at a pressure of 600 MPa for 1 min significantly reduced PME activity in UBMP and significantly activated PME in BMP, whereas pressures of 300–500 MPa activated PME regardless of blanching. However, PME activity was reduced by 97 % in UBMP and was completely inactivated in BMP by HTST treatment. WSP levels were significantly decreased by HHP treatment but were increased by HTST treatment in UBMP and BMP. Both HHP and HTST treatments increased the viscosity, storage modulus, and loss modulus of UBMP and BMP. No significant changes in total sugar, total soluble solids, titratable acid, or pH were found after any treatment.     

Effects of high pressure treatment on the quality and storage of kimchi

The effects of high pressure treatment on the microflora and storage of kimchi were investigated. In a bacterial suspension, numbers of Lactobacillus plantarum were reduced by 6 logs by 500 MPa, at 25 °C for 10 min. Kimchi juice did not alter the rate of inactivation of lactic acid bacteria by high pressure treatment. There was no change in the texture of kimchi subjected to a pressure of 400 MPa, but an increase in cutting force was observed at 600 MPa. When kimchi was pressurized at 400 MPa for 10 min at 25 °C and subsequently stored at 20 °C for 4 weeks, the total number of viable cells stayed at 10³ CFU mL⁻¹. High pressure treatment above 400 MPa prevented excessive acidification that typically occurs during the extended storage of kimchi. The inflation of pouches as a result of accumulated carbon dioxide was also prevented by high pressure treatment. Although colour changes were accelerated by high pressure treatment, this study demonstrates that high pressure treatment can be used to control overripening during the distribution and storage of kimchi products.     

Evaluation of processing qualities of tomato juice induced by thermal and pressure processing

Effects of processing conditions including hot-break processing (92 °C for 2 min), cold-break processing (60 °C for 2 min) and hydrostatic pressure treatments (100-500 MPa) at different temperatures (4, 25 and 50 °C) for 10 min on quality aspects of tomato juice were investigated. Both hot- and cold-break processing induced significant changes in color, viscosity and radical-scavenging capacity of tomato juice compared with control (fresh tomato juice); moreover, hot-break processing induced a specific range of reduction of pectin methylesterase (PME) and polygalacturonase (PG) activities. Pressure treatments at and below 200 MPa at 4 and 25 °C maintained the color, extractable total carotenoids and lycopene, and radical-scavenging capacity; further, those at 500 MPa at 4 and 25 °C improved all the quality attributes the most except inactivation of PME in this study. The residual activity of PME showed the lowest after treating by 200 MPa at 25 °C; however, the PME activity was enhanced by treatments at 300-500 MPa and various temperatures. The residual activity of PG decreased gradually to 72% with pressure elevated from 100 to 400 MPa at 4 and 25 °C, further, that declined quickly to 10% after 500 MPa treatments. This research clearly shows that it is possible to selectively produce good tomato juice products by high pressure processing at ambient temperature.     

Comparison of high pressure treatment and thermal pasteurization effects on the quality and shelf life of guava puree

The effects of high pressures and thermal pasteurization on the survival of microorganisms, enzyme inactivation and quality changes of guava puree during storage at 4°C were investigated and compared with untreated samples. After treatment at a pressure of 600MPa and 25°C for 15 min, the microorganisms in guava puree were inactivated to less than 10 cfu mL⁻¹ and the product exhibited no change in colour, pectin, cloud and ascorbic acid content as compared with fresh samples. The inactivation of enzymes in guava puree by thermal pasteurization was greater than by high pressures. The microbial count in guava puree reduced to 200 cfu mL⁻¹ and the product showed marked changes in viscosity, turbidity and colour when heated at 88–90°C for 24s. The content of pectin, cloud and ascorbic acid as well as colour in untreated and high pressurized (400MPa) guava puree gradually decreased, whereas these changes were not observed in pasteurized (88–90°C) and high pressurized (6000MPa) puree during storage at 4°C for 60 days. The guava puree treated at 600MPa and 25°C for 15 min retained good quality similar to the freshly extracted puree after storage at 4°C for 40 days.     

Modeling the inactivation of pectin methylesterase in pineapple puree during combined high-pressure and temperature treatments

The inactivation of pectin methylesterase (PME) in pineapple puree was studied within the domain of 0.1–600 MPa/30–70 °C/1 s–40 min. The combined effect of pressure-build up and decompression, as characterized by pulse inactivation (PI value), was modeled by the artificial neural network (ANN) through a tan-sigmoidal function of target pressure, target temperature, compression, and decompression time. Besides, nth order kinetic model was fitted during the isobaric-isothermal hold period. The extent of pulse inactivation of PME ranged from 15% (200 MPa/30 °C) to 67% (600 MPa/70 °C) and it increased at a higher temperature and/or pressure. The inactivation orders (n) during thermal (0.1 MPa/30–70 °C) and high pressure (100–600 MPa/30–70 °C) treatments were 1.15 and 1.3, respectively. The rate constant (k) ranged within 4.0 to 71.2 × 10⁻³ Uⁿ⁻¹·min⁻¹. A nonlinear model considering the pressure dependency of activation energy, and temperature dependency of activation volume was developed which adequately described the inactivation behavior of PME within the domain.Industrial relevancePectin methylesterase (PME) in the pineapple puree results in a product with a modified texture and consistency that is usually not entertained by the consumer. Therefore, pineapple puree has to be processed to inactivate PME to avoid the cloud loss. Now-a-days, high-pressure processing is being used for fruit products to retain the heat sensitive nutrients. In this sense, a model capable of predicting the exact inactivation behavior of PME during the treatment is very much obligatory for process design. This combined model developed in the study will help the food industry to come-up with the exact pressure-temperature-holding time combination achieving a certain degree of PME inactivation.     

Kinetics of Tomato Pectin Methylesterase Inactivation by Temperature and High Pressure

ABSTRACT: In this study we investigated the inactivation of endogenous pectin methylesterase (PME) in tomato juice during combined high-hydrostatic pressure (ambient to 800 MPa) and moderate temperature (60 to 75 °C) treatments under isobaric and isothermal processing conditions. PME inactivation rates increased with increasing processing temperature, with the highest rate obtained during processing at 75 °C and ambient pressure. Inactivation rates were dramatically reduced as soon as processing pressure was raised. High inactivation rates were again attained when processing pressure exceeded a value of about 700 MPa. Such a behavior was described by considering two parallel mechanisms of inactivation, each one following first order kinetics with its own kinetic parameters.     

高静压对桃汁杀菌、钝化酶活性的效果

研究在不同处理压力和时间条件下,高静压加工技术对桃汁中微生物(细菌总数、霉菌、酵母菌、大肠菌群)以及酶(多酚氧化酶、果胶甲基酯酶、脂肪氧化酶)的影响。结果表明:经400MPa、5min高静压处理即可完全杀灭桃汁中的微生物;在400MPa和500MPa条件下,桃汁中的多酚氧化酶和脂肪氧化酶的活性出现了不同程度的激活现象,但在600MPa时,随着处理时间的延长,其活性逐渐降低,经30min处理后,分别被钝化了0.7662和0.641。而果胶甲基酯酶在400、500、600MPa条件下,出现了不规律的激活或钝化现象。另外,研究表明在高静压加工前增加漂烫工艺,可以有效杀灭桃汁中的微生物及钝化酶活性。     

Process stability of Capsicum annuum pectin methylesterase in model systems, pepper puree and intact pepper tissue

Process stability studies towards temperature and/or pressure on pepper pectin methylesterase (PME) were carried out in different systems (purified form, crude extract, pepper pieces and puree) at pH 5.6. Within the temperature range studied (22–80 °C, 5 min), pepper PME in pure form and crude extract was gradually inactivated showing a biphasic inactivation behaviour, indicating the presence of isoenzymes of different thermostability. Pepper samples heated for 15 min showed a maximum of residual PME activity around 55 °C. Isothermal inactivation of pepper PME in purified form and crude extract at pH 5.6 could be described by a biphasic inactivation model for the temperature range studied (62–76 °C). A stable behaviour towards high-pressure/temperature treatments (400–800 MPa/25–60 °C) was observed for crude extract and purified pepper PME. PME in pepper puree samples revealed to be very pressure stable. Mild temperatures combined with pressure treatments seem to increase the extractability from PME in pepper tissue, probably due to the effect on the cell structure.     

Inactivation of Pectin Methyl Esterase in Orange Juice by Pulsed Electric Fields

ABSTRACT: Pulsed electric fields (PEF) treatments were applied to nonpasteurized orange juice using a bench top PEF system to study effects of PEF on the activity of pectin methyl esterase (PME). Effects of electric strength on PME activity at a constant water bath temperature were studied using electric field strengths up to 35 kV/cm at 30 °C. Increase of electric field strength caused a significant inactivation of PME with increase in orange juice temperature ( p < 0.05). A thermal inactivation study showed that heating of orange juice at the same temperature as orange juice during PEF treatment was not effective as PEF treatment in inactivating PME. Effects of electric field strength at different water bath temperatures were studied using electric field strengths up to 25 kV/cm and water bath temperatures of 10–50 °C. Higher electric field strengths at higher water bath temperature were the more effective to inactivate PME. A combination of PEF treatment at 25 kV/cm and a water bath temperature of 50 °C caused 90% inactivation of PME.     

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.     

THERMAL AND HIGH-PRESSURE STABILITY OF PURIFIED PECTIN METHYLESTERASE FROM PLUMS (PRUNUS DOMESTICA)

Pectin methylesterase (PME) from greengage plums (Prunus domestica) has been extracted and purified using affinity chromatography. Only one band on sodium dodecyl sulfate–polyacrylamide gel electrophoresis was obtained, with an estimated molecular weight of 31 kDa. On isoelectric focusing electrophoresis, two bands with neutral isoelectric points (6.8 and 7.0) were detected. The optimal pH and temperature for plum PME activity were 7.5 and 65C, respectively. A study of purified plum PME thermostability was performed at pH 7.5 and 4.0, indicating a higher thermostability at pH 7.5 than at pH 4.0. A biphasic inactivation behavior was observed for thermal treatments (54–70C), whereas its pressure inactivation could be described by a first‐order kinetic model in a pressure range of 650–800 MPa at 25C. Purified plum PME was found to be relatively stable to thermal and pressure (≤600 MPa) treatments, compared to PME from other fruits.     

High Pressure and Temperature Effects on Enzyme Inactivation in Strawberry and Orange Products

High hydrostatic pressure treatment (50-400 MPa) combined with heat treatment (20–60°C) effects on peroxidase (POD), polyphenoloxidase (PPO) and pectin methylesterase (PME) activities of fruit-derived products were studied. Assays were carried out on fresh orange juice and strawberry puree. Pressurization/depressurization treatments caused a significant loss of strawberry PPO (60%) up to 250 MPa and POD activity (25%) up to 230 MPa, while some activation was observed for treatments carried out in 250–400 MPa range for both enzymes. Optimal inactivation of POD was using 230 Mpa and 43°C in strawberry puree. Combinations of high pressure and temperature effectively reduced POD activity in orange juice (50%) to 35°C. The effects of high pressure and temperature on PME activity in orange juice were very similar to those for POD.     

Pressure Inactivation Kinetics of Microbial Transglutaminase from Streptoverticillium mobaraense

Microbial transglutaminase (MTGase) forms nondisulfide covalent crosslinks in proteins. Changes in activity of MTGase from Streptoverticillium mobaraense after exposure to pressure (100 to 600 MPa for 10 to 60 min, respectively) were analyzed. MTGase activity increased linearly with enzyme concentration, regardless of pressures in the range examined. The Vmax value was changed by pressure while the Km value was independent of pressure. Sodium chloride (0 to 0.86 N) apparently caused MTGase destabilization under pressure. Oscillatory pressurization (400 to 600 MPa, 25°C, 10 min/cycle, 0 to 5 cycles) resulted in a higher degree of inactivation when pressures were greater than 500 MPa. MTGase maintained 60% of its initial activity even after pressurization at 600 MPa for 60 min, indicating that MTGase was pressure‐resistant as compared to other enzymes.     

Enzyme Activity and Nutritional Quality of Peach (Prunus persica) Juice: Effect of High Hydrostatic Pressure

The inactivation of polyphenol oxidase and pectin methylesterase in peach juice was investigated after high hydrostatic pressure processing at 400–600 MPa and 25°C for 5–25 min, respectively. At 400 MPa, polyphenol oxidase and pectin methylesterase were activated by 7.3 and 2.6%. At 500 and 600 MPa, polyphenol oxidase and pectin methylesterase were inactivated significantly with increasing the pressure and time, and the inactivation kinetics was fitted by the first order model. Moreover, some physio-chemical properties were studied. The results revealed that high hydrostatic pressure treatment preserved more L-ascorbic acid and maintained the color and sensory quality better than thermal treatment.     

超高压处理对鲜橙汁中果胶酶及过氧化物酶活性的影响

为推进国内非冷冻浓缩橙汁加工业的发展,对新型超高压杀菌技术对橙汁中酶钝化的效果进行研究,采用200～600MPa超高静压处理鲜榨橙汁,使用紫外分光光度法、滴定法分别测定鲜橙汁中过氧化物酶(POD)和果胶酶(PME)的活性,进行两种酶在常温下超高压钝化酶一级动力学拟合研究。结果表明：在常温条件下200MPa处理10min使两种酶轻微激活,在300～600MPa条件下,随压力和处理时间的增大,两种酶钝化反应明显,且符合一级动力学模型,且果胶酶对压力钝化更加敏感。     

Effect of high hydrostatic pressure on Listeria innocua 910 CECT inoculated into Ewe's milk.

Ewe's milk standardized to 6% fat was inoculated with Listeria innocua 910 CECT at a concentration of 10(7)CFU/ml and treated by high hydrostatic pressure. Treatments consisted of combinations of pressure (200, 300, 350, 400, 450, and 500 MPa), temperature (2, 10, 25, and 50 degrees C), and time (5, 10, and 15 min). To determine numbers of L. innocua, listeria selective agar base with listeria selective supplement and plate count agar was used. Low-temperature (2 degrees C) pressurizations produced higher L. innocua inactivation than treatments at room temperatures (25 degrees C). Pressures between 450 and 500 MPa for 10 to 15 min were needed to achieve reductions of 7 to 8 log units. The kinetics of destruction of L. innocua were first order with D-values of 3.12 min at 2 degrees C and 400 MPa and 4 min at 25 degrees C and 400 MPa. A baroprotective effect of ewe's milk (6% fat) on L. innocua was observed in comparison with other studies using different media and similar pressurization conditions.     

Thermal and High-Pressure Stability of Pectin-Converting Enzymes in Broccoli and Carrot Purée: Towards the Creation of Specific Endogenous Enzyme Populations Through Processing

The thermal and pressure stability of broccoli and carrot pectin-converting enzymes, in particular pectinmethylesterase (PME), β-galactosidase (β-Gal), and α-arabinofuranosidase (α-Af), were investigated in vegetable purée matrices. In situ enzyme inactivation by thermal and high-pressure processing (respectively 5 min at 25–80 °C at 0.1 MPa and 10 min at 0.1–800 MPa at 20 °C) was evaluated by measuring the residual enzyme activity in crude extracts of treated carrot, broccoli floret, and broccoli stem purée samples. PME was completely inactivated in all vegetable purée matrices after a 5-min treatment at 80 °C. After a treatment at 800 MPa (20 °C, 10 min) only 77–90 % of pressure stable PME was inactivated, depending on the matrix. β-Gal and α-Af enzymes were inactivated in the vegetable purée matrices by thermal treatments respectively at 67.5–72.5 and 80 °C. These enzymes showed some pressure resistance: treatments respectively at 600–700 and 600–750 MPa were necessary for one log-reduction of β-Gal and α-Af activity in the different purées at 20 °C. Under the assumption of a first-order inactivation model, inactivation rate constants and their temperature or pressure dependency were determined for the different enzymes. Based on differences in process stability of the enzymes in the individual purée matrices, the feasibility for the creation of specific endogenous enzyme populations by selective processing was evaluated.     

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.