Kinetic study of high pressure microbial and enzyme inactivation and selection of pasteurisation conditions for Valencia Orange Juice

Keeping quality of fresh orange juice is highly dependent on pectinolytic enzyme activity and the growth of spoilage microorganisms. The inactivation kinetics of indigenous pectin methylesterase (PME) and of the two more pressure resistant species of spoilage lactic acid bacteria (LAB) Lactobacillus plantarum and L. brevis in freshly squeezed Valencia orange juice under high hydrostatic pressure (100–500 MPa) combined with moderate temperature (20–40 °C) was investigated. PME inactivation followed first order kinetics with a residual PME activity (15%) at all pressure–temperature combinations used. The values of activation energy and activation volume were estimated at each pressure and at each temperature, respectively. Values of 90 kJ/mol and ?30 mL/mol at reference pressure of 300 MPa and reference temperature of 35 °C were estimated respectively. The corresponding z_T and z_P values of LAB inactivation were also estimated at all conditions tested. Values of 19.5 °C and 95 MPa at reference pressure of 300 MPa and reference temperature of 30 °C were estimated respectively for L. plantarum, while the corresponding values for L. brevis were 40 °C and 82 MPa, respectively, at the same reference conditions. Pressure and temperature were found to act synergistically both for PME and LAB inactivation. The PME and LAB inactivation rate constants were expressed as functions of the temperature and pressure process conditions. These functions allow the determination of the pressure/temperature conditions that achieve the target enzyme and microbial inactivation at a selected processing time. The process conditions of 350 MPa at 35 °C for 2 min are proposed as effective for Valencia orange juice cold pasteurisation.     

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.     

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.     

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.     

Thermal and High-Pressure Stability of Pectinmethylesterase, Polygalacturonase, β-Galactosidase and α-Arabinofuranosidase in a Tomato Matrix: Towards the Creation of Specific Endogenous Enzyme Populations Through Processing

The thermal and pressure stability of tomato pectinmethylesterase (PME), polygalacturonase (PG), β-galactosidase (β-Gal), and α-arabinofuranosidase (α-Af) were investigated in situ. Enzyme inactivation by thermal and high-pressure processing (respectively 5 min at 25–95 °C at 0.1 MPa and 10 min at 0.1–800 MPa at 20 °C) was monitored by measuring the residual activity in crude enzyme extracts of treated tomato purée samples. PME was completely inactivated after a 5-min treatment at 75 °C. Only 30 % of the pressure stable PME was inactivated after a treatment at 800 MPa (20 °C, 10 min). A 5-min treatment at 95 °C and a treatment at 550 MPa (20 °C, 10 min) caused complete PG inactivation. β-Gal and α-Af activities were already reduced significantly by thermal treatments at 42.5–52.5 °C and 45–60 °C, respectively. These enzymes were, however, rather pressure resistant: treatments at respectively 700 and 600 MPa were necessary to reduce the activity below 10 % of the initial value. Assuming that first-order, fractional conversion or biphasic inactivation models could be applied to the respective enzyme inactivation data, inactivation rate constants and their temperature or pressure dependence for the different enzymes were determined. Based on differences in process stability of the enzymes, possibilities for the creation of specific “enzyme populations” in tomato purée by selective enzyme inactivation were identified. For industrially relevant process conditions, the enzyme inactivation data obtained for tomato purée were shown to be transferable to intact tomato tissue.     

Optimisation of High Hydrostatic Pressure Processing of Pêra Rio Orange Juice

The influence of high hydrostatic pressure (HHP) on Pêra Rio orange juice was investigated using response surface methodology. A central composite design was used to evaluate the effects of three processing conditions (independent variables), namely pressure (100–600 MPa), temperature (30–60 °C) and time (30–360 s), on the native microflora and pectin methylesterase (PME) activity of orange juice. Analysis of variance showed that second-order polynomial models fitted well with the experimental data for PME residual activity (R ²?=?0.9586, p?<?0.001) and aerobic microorganism count (R ²?=?0.9879, p?<?0.001). The optimum HHP processing conditions to produce orange juice with PME residual activity of less than 20 % and low microorganism count (<2 log cycles CFU/mL) were 550 to 600 MPa, 55 to 60 °C and 330 to 360 s.     

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.     

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.     

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.     

Inactivation Kinetics of Peach Pulp Pectin Methylesterase as a Function of High Hydrostatic Pressure and Temperature Process Conditions

A kinetic study of the inactivation of endogenous pectin methylesterase (PME) in Greek commercial peach pulp under high hydrostatic pressure (HHP; 100–800 MPa) combined with moderate temperature (30–70 °C) was conducted. Thermal inactivation of the enzyme at ambient pressure conditions was also studied. PME inactivation was modeled by first order kinetics at all conditions tested. High pressure and temperature acted synergistically on PME inactivation, except at the high temperature of 70 °C at the middle pressure range (100–600 MPa), where an antagonistic effect of pressure and temperature was observed. At this specific middle pressure range, an increase of pressure processing led to increased inactivation rate constants of peach PME. A multiparameter model was developed to express the PME inactivation rate constant as a function of temperature and pressure process conditions, taking into account the dependence of both activation energy and activation volume on pressure and temperature, respectively. A good correlation between experimental and predicted values of inactivation rate constants was established. This modeling approach enables the quantitative estimation of the HHP–temperature conditions needed to achieve a targeted PME inactivation in the peach pulp.     

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。     

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.     

Inactivation Kinetics of Pectin Methylesterase,Polyphenol Oxidase,and Peroxidase in Cloudy Apple Juice under Microwave and Conventional Heating to Evaluate Non-Thermal Microwave Effects

Continuous-flow microwave pasteurization provides important advantages over conventional heat exchangers such as fast volumetric heating, lower tube surface temperature, and possible non-thermal effects that enhance enzymatic and bacterial inactivation. Conventional and microwave-assisted inactivation of pectin methylesterase (PME), polyphenol oxidase (PPO), and peroxidase (POD) in cloudy apple juice were investigated to evaluate non-thermal effects. Experiments were conducted to provide uniform heating with accurate temperature acquisition and similar temperature profiles for conventional and microwave treatments. A two-fraction first-order kinetic model was successfully fitted to the data in a procedure that took into account the whole time-temperature profile instead of assuming isothermal conditions. Predicted inactivation curves for pasteurization at 70 and 80 °C of the cloudy apple juice showed that PME has the highest thermal resistance (residual activity of 30% after 250 s at 80 °C) and that there was no evidence of non-thermal microwave effects on the inactivation of these enzymes.     

Pectinesterase Activity and Proximate Analyses of Sea Buckthorn Juices

Three varieties of sea buckthorn fruit were harvested and pressed to obtain juices. These were analyzed for pectin methylesterase activity, moisture, nitrogen, oil, pH, total acid, and °Brix. Yields of press juice for Hippophae rhamnoides ssp. rhamnoides L. varieties Luchistaya, Prozrachnaya, and ssp. mongolica Rousi cv. Indian Summer were 68%, 69%, and 66% w/w, respectively. Differences in juice composition were found to be in moisture content, °Brix, pH, and in total acid with the Luchistaya variety having the highest acid levels and lowest pH. Pectin methylesterase (PME) was present in all sea buckthorn juices with initial activity levels of 6.8 to 14.0 μ equivalents per min per 100 g of juice at pH 8 and 23 °C. Activity was pH‐dependent with little PME activity at pH 3 to 5 and the highest activity at pH 8. The cream, pellet, and serum layers of centrifuged juices all contained PME. Heat treatment reduced PME activity in the juices by 1 decimal reduction or less.     

Novel,non-thermal hydrodynamic cavitation of orange juice: Effects on physical properties and stability of bioactive compounds

Effect of hydrodynamic cavitation (HC) at pressure 3–15 bar and treatment time5–25 min on physical and chemical qualities of orange juice was investigated. Processing parameters were optimized on the basis of retention of vitamin C and antioxidant activity; inactivation of pectin methyl esterases (PME) and peroxidase (POD) enzyme and stability of pH, °Brix, viscosity, titratable acidity and total color difference. HC of orange juice at 5 bar 15 min and 13 bar 10 min resulted into maximal overall desirability at 0.52 and 0.40 respectively. No significant change in °Brix, pH, titratable acidity for fresh and cavitated orange juice was observed. Only 21% and 13% of reduction in PME and POD respectively was recorded. A 94% and 91% of antioxidant activity and vitamin C retention was noted in both optimized samples. This study demonstrated that HC can produce orange juice more economically with better physical properties and nutritional value.Industrial relevanceThe demand for more natural, preservative free with highest nutritional bioactive containing juices has created a need of non-thermal processing. Hydrodynamic cavitation (HC) processing is novel, emerging and under explored technology for fruit juices. HC due to the formation of cavities and shock waves causes enzyme and microbial inactivation at low temperature while retaining natural bioactives with fresh-like organoleptic characteristics. It is believed worldwide that non thermal technologies like HC will be among the most cost effective, scalable and impactful liquid food processing technologies in the coming decades especially for commercial products.     

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.     

High-Pressure Processing of Orange Juice: Kinetics of Pectinmethylesterase Inactivation

ABSTRACT: A kinetic study of pectinmethylesterase (PME) inactivation in orange juice was conducted. Juice samples were subjected to combinations of high pressure (400, 500, 600 MPa) and thermal (25, 37.5, 50 °C) treatments for various time periods. PME inactivation followed a first-order kinetic model with a residual activity of pressure-resistant enzyme remaining. Calculated D-values ranged from 4.6 min to 117.5 min at 600 MPa/50 °C and 400 MPa/25 °C, respectively. Pressures in excess of 500 MPa resulted in sufficiently fast inactivation rates for economic viability of the process.     

Vitamin C Content in Sea Buckthorn Berries (Hippophaë rhamnoides L. ssp. rhamnoides) and Related Products: A Kinetic Study on Storage Stability and the Determination of Processing Effects

ABSTRACT: The primary vitamin in sea buckthorn berries is vitamin C containing values of approximately 400 mg/100 g. Processing effects were investigated during juice and concentrate production from sea buckthorn berries (Hippophaë rhamnoides) and storage stability of juices was determined for up to 7 d using berries and juices from 2 different growing areas. During industrial juice production the technological processing of the berries caused a loss of about 5% to 11% total ascorbic acid (TAA) in the generated juice. The production of the concentrated juice resulted in 50% depletion of TAA. Sea buckthorn berries and juice were stored at 6, 25, and 40 °C for up to 7 d to investigate the temperature effects on TAA during storage. Analysis of kinetic data suggested that the degradation follows a 1st‐order model. The results of the experiments showed that storage of sea buckthorn juices for 7 d at cold temperature (6 °C) already resulted in a degradation of TAA of about 11% to 12%.     

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

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