Impact of pulsed light treatment on enzyme inactivation and quality attributes of whole white button mushroom (Agaricus bisporus) and its storage study.

Whole white button mushrooms (WWBM) exhibit a limited shelf-life owing to the oxidative enzymatic browning. Inactivation of polyphenol oxidase-PPO and peroxidase-POD in WWBM and its kinetic behavior were studied using pulsed light(PL) treatment (0.13-1.11 J/cm2). The first-order kinetics explained PL-induced enzyme inactivation. Rate constants(k) for PPO and POD were 3.84 and 2.55 cm2/J. FTIR-analysis revealed secondary-structural changes in partially-purified enzyme. PL-treatment retarded browning, retained phenolics and enhanced vitamin D2. PL-treatment at 1.11 J/cm2 rendered WWBM both microbially and enzymatically stable. The PL-treated WWBM's shelf-life at 4, 20, and 37 °C were 5, 3, and 1 day. At 4 °C, browning increased by 6.1 %; firmness decreased by 55.2 %, while PL-treated mushrooms retained 90.6 % phenolics, 78.9 % antioxidant capacity, and 64.2 % D2 after 5 days. Higher activation energy value confirmed phenolics were most sensitive during storage. PL-technology supports UN Sustainable Development Goals by reducing chemical use, lowering carbon-footprints, minimizing pollution, and enhancing shelf-life, promoting sustainable global trade.

Variations in quality attributes of pulsed light-treated table grape juice during refrigerated storage (4°C) and ambient conditions (25°C).

Pulsed light (PL) pasteurization is being explored as a substitute for the conventional thermal pasteurization of juices in recent times due to better retention of nutrients and overall quality. However, the long-term stability of the PL-pasteurized juice must be investigated to promote its application by the industry. The effect of PL treatment (effective fluence of 1.15 J·cm-2) and thermal treatment (90°C for 60 s) on microbial quality, enzyme activity, bioactive compounds, sensory acceptance, and color profile of table grape juice during storage at 4 and 25°C was investigated in this study. The PL pasteurization enhanced the microbial shelf-life of the juice (<6 log10cfu·mL-1) from 5 to 35 days at 4°C. The PL and thermally-pasteurized juice demonstrated a shelf-life of only 10 days when stored at 25°C. The total soluble solids and titratable acidity did not alter significantly throughout the storage period. The peroxidase, polyphenol oxidase, and pectin methylesterase activities were below 10% for the PL and thermally-treated beverage when stored at 4°C. The sensory acceptability of the PL-pasteurized juice after 35 days of refrigerated storage (6.9 ± 0.3) was close to the untreated juice (7.2 ± 0.3) and greater than thermally-treated juice (6.2 ± 0.2). After the 35th day of storage at 4°C, PL-treated grape juice retained 55%, 12%, and 15.3% more phenolics, flavonoids, and antioxidant capacity, respectively, than the thermally-pasteurized juice. Hence, PL pasteurization can effectively prolong the shelf-life of table grape juice while achieving microbial and enzymatic stability, along with high sensory and nutritional appeal. PRACTICAL APPLICATION: Exploring non-thermal methods like pulsed light (PL) pasteurization as a substitute for conventional thermal methods is gaining recognition for its ability to retain nutrients and improve overall juice quality. However, the industry's adoption depends on understanding the shelf-stability of PL-pasteurized juice. This study specifically investigates the practical applications of PL treatment in comparison with conventional thermal treatment in enhancing microbial safety and enzymatic stability in table grape juice. The findings contribute insights into optimizing the shelf life of table grape juice and preserving its quality, supported by microbial, enzymatic, and sensory evaluations.

Sequential Pulsed Light and Ultrasound Treatments for the Inactivation of Saccharomyces cerevisiae and PPO and the Retention of Bioactive Compounds in Sweet Lime Juice.

Designing a pasteurization con dition for sweet lime juice while ensuring microbial safety, enzymatic stability, and high nutritional quality is crucial for satisfying stakeholder demands. The present research investigates the effects of matrix pH, ultrasound treatments, and sequential pulsed light on the microbial population, enzyme activity, and bioactive chemicals in sweet lime juice. The sequential pulsed light (PL: 0.6-0.84 J/cm2) and ultrasound (US: 0.2-0.4 W/cm3) treatments for sweet lime juice were optimized using response surface methodology (RSM). A three-factor full factorial design was used for this purpose. The independent variables encompassed pH (X1), PL effective fluence (X2, J/cm2), and US intensity (X3, W/cm3). The responses assessed included the inactivation of Saccharomyces cerevisiae (Y1, log cfu/mL) and polyphenol oxidase (PPO: Y2 in %) and the retention of vitamin C (Y3, %). The polynomial models were optimized using numerical optimization to attain the maximum desirability value (0.89). The optimized PL + US sample (0.8 J/cm2 + 0.4 W/cm3, respectively) at pH 3.5 resulted in a 5-log cycle reduction in S. cerevisiae count and a 90% inactivation in PPO activity and retained 95% of its vitamin C content. This optimized sample underwent further analysis, including phenolic profiling, assessment of microbial cell morphology, and examination of enzyme conformational changes. After sequential pulsed-light (0.8 J/cm2) and ultrasound (0.4 W/cm3) treatments, yeast cells showed unusual structural changes, indicating additional targets besides membranes. Following PL + US treatment, the PPO composition changed to 2.7 ± 0.1% α-helix, 33.9 ± 0.3% ß-sheet, 1.4 ± 0.2% ß-turn, and 62 ± 0.7% random coil. Impressively, the optimized PL + US sample maintained a sensory acceptance level similar to that of the untreated sample.

Pulsed light treatment of whole white button mushroom (Agaricus bisporus): Kinetics and mechanism of microbial inactivation and storage study.

The whole white button mushrooms (WWBMs) are highly perishable due to susceptibility to microbial spoilage. This study explored the potential of pulsed light (PL) treatment for decontamination and shelf-life extension of WWBM. WWBM surface was inoculated with Escherichia coli, Listeria monocytogenes, and Aspergillus niger spores (8.1, 8.0, and 8.05 log10 CFU/g, respectively) and tested for inactivation against various PL intensities (fluence 0.13-0.75 J/cm2). The kinetics and mechanism of microbial inactivation were explored, and shelf life was determined at 4, 20, and 37°C. Microbial inactivation increased with increasing PL intensity. PL-induced microbial inactivation was well explained by Weibull model with shape parameters (ß-value) for E. coli, L. monocytogenes, A. niger, aerobic mesophiles, and yeast and mold as 0.87, 0.92, 0.91, 0.89, and 0.94, respectively. PL-treatment at 0.75 J/cm2 resulted in >5-log cycle reduction in all inoculated and natural microorganisms. Exposure to PL led to collapse of cellular structure, ruptured cell wall, and leakage of cellular material in all microorganisms and spores along with alterations in nucleic acid and lipid bands. At 4°C, maximum shelf life of 5 days was achieved when WWBM was exposed at 0.75 J/cm2. The WWBM retained 83.3% phenolics, 83.9% antioxidant capacity, and 77.4% vitamin D2 at 4°C while reducing the polyphenol oxidase and peroxidase activity by 89% and 79%. The degradation rate for quality parameters increased with storage temperature. The activation energy of the browning index affirmed it as the most sensitive quality attribute during storage. The study concluded the potential of PL treatment to prolong the shelf life of WWBM.

Advances in the novel and green-assisted techniques for extraction of bioactive compounds from millets: A comprehensive review.

Millets are rich in nutritional and bioactive compounds, including polyphenols and flavonoids, and have the potential to combat malnutrition and various diseases. However, extracting these bioactive compounds can be challenging, as conventional methods are energy-intensive and can lead to thermal degradation. Green-assisted techniques have emerged as promising methods for sustainable and efficient extraction. This review explores recent trends in employing green-assisted techniques for extracting bioactive compounds from millets, and potential applications in the food and pharmaceutical industries. The objective is to evaluate and comprehend the parameters involved in different extraction methods, including energy efficiency, extraction yield, and the preservation of compound quality. The potential synergies achieved by integrating multiple extraction methods, and optimizing extraction efficiency for millet applications are also discussed. Among several, Ultrasound and Microwave-assisted extraction stand out for their rapidity, although there is a need for further research in the context of minor millets. Enzyme-assisted extraction, with its low energy input and ability to handle complex matrices, holds significant potential. Pulsed electric field-assisted extraction, despite being a non-thermal approach, requires further optimization for millet-specific applications, are few highlights. The review emphasizes the importance of considering specific compound characteristics, extraction efficiency, purity requirements, and operational costs when selecting an ideal technique. Ongoing research aims to optimize novel extraction processes for millets and their byproducts, offering promising applications in the development of millet-based nutraceutical food products. Therefore, the current study benefits researchers and industries to advance extraction research and develop efficient, sustainable, and scalable techniques to extract bioactive compounds from millets.

Effect of continuous microwave processing on enzymes and quality attributes of bael beverage.

Bael (Aegle marmelos) beverage was pasteurized using continuous-microwave (MW) and traditional thermal processing and the activity of native enzymes, pulp-hydrolyzing enzymes, bioactive, physicochemical, and sensory properties were analyzed. First-order and linear biphasic models fitted well (R2 ≥ 0.90) for enzyme inactivation and bioactive alteration kinetics, respectively. For the most resistant enzyme, polyphenoloxidase (PPO), the inactivation target of ≥ 90 % was achieved at 90 °C TMW (final temperature under MW) and 95 °C for 5 min (conventional thermal). MW treatment displayed faster enzyme inactivation and better retention of TPC and AOC. MW treatment at 90 °C TMW showed 5.3 min D-value, 90% total carotenoid content, 3.42 crisp sensory score (out of 5), and no or minor change in physicochemical attributes. Thermal and MW treatment caused the loss of 14 and 10 bioactive compounds, respectively. The secondary and tertiary structural modifications of PPO enzyme-protein revealed MW's lethality primarily due to its thermal effects.

Formulation of a mixed fruit beverage using fuzzy logic optimization of sensory data and designing its batch thermal pasteurization process.

The study aims to formulate a mixed fruit beverage through sensory analysis, and the composition was optimized using a fuzzy logic algorithm. The fuzzy optimization algorithm was developed using a modified Takagi and Sugeno's approach, polynomial mixture modeling, and nonlinear solver engine. The optimized blend consisted of amla juice, pineapple juice, and coconut water in 14.3, 63.0, and 22.7%, respectively. Further, the batch thermal treatment was carried out within 50 to 95 °C for an isothermal holding time of 1 s to 10 min, and pasteurization condition for the beverage was estimated from kinetic modeling. The concept of thermal pulse inactivation due to non-isothermal heat-up-time and cool-down-time has been introduced within the process time calculation. From the kinetic study, polyphenoloxidase enzyme appeared to be the most resistant entity towards inactivation among all the natural microbiota and quality deteriorating enzymes. Pasteurization in terms of achieving a 5D reduction of both aerobic mesophilic and yeast-mold counts was attained over a range of 80 to 95 °C for 10.2 + 1.4 to 3.1 + 2.0 min (1.4 and 2.0 min = heat-up-time + cool-down-time), respectively. The 90% inactivation of both polyphenoloxidase and peroxidase enzymes was obtained over a range of 90 to 95 °C for 12.8 + 1.7 to 8.4 + 2.0 min, respectively. While obtaining both the microbial and enzyme stability at the isothermal condition of 95 °C for 8.4 min, the corresponding retention in ascorbic acid, total phenolics, and antioxidant capacity were observed as 49.7, 63.0, and 61.4%, respectively. PRACTICAL APPLICATION: In this work, the formulation of a fruit blend was optimized through an intelligent optimization technique (fuzzy algorithm) applied to the sensory data set. The approach for calculating thermal processing time or pasteurization condition provides a new dimension with better precision. The thermal treatment condition of 95 °C for 10 min can be used for this mixed beverage to achieve both microbial stability (5-log reduction) and enzyme stability (90% reduction). The presented study can be used as a reference for other similar beverages to achieve a complete process design from basic formulation optimization to thermal (batch-type) processing conditions.

Partial purification, characterisation and thermal inactivation kinetics of peroxidase and polyphenol oxidase isolated from Kalipatti sapota (Manilkara zapota).

BACKGROUND: The extraction, purification, and characterisation of peroxidase (POD) and polyphenol oxidase (PPO) were studied for Kalipatti sapota fruit. The crude enzyme extract was partially purified by ammonium sulfate precipitation followed by BioGel P100 size exclusion and Unosphere Q anion-exchange chromatography. RESULTS: Molecular weights of 20 kDa (POD) and 24 kDa (PPO) were indicated by SDS-PAGE. A single band was observed on SDS-PAGE with a fold purity of 10.38 and 7.42 for POD and PPO, respectively. Michaelis-Menten constants for POD and PPO were 22.3 and 23.0 mmol L-1 using guaiacol and catechol as substrates. Thermal inactivation kinetics was studied in the temperature range of 60-95 °C. The crude extract of POD and PPO showed D-values of 2.2-60.2 and 1.0-35.2 min; Z-values of 18.7 ± 0.4 and 16.0 ± 0.3 °C; and activation energies (Ea ) of 128.6 and 151.0 kJ mol-1 , respectively. CONCLUSION: POD and PPO showed good stability over a wide range of pH and temperature. As reflected by Z and Ea values, the fruit matrix had no significant influence towards enzyme stability. Designing of thermal process should take into consideration D- and Z-values of the enzymes along with D- and Z-values of microorganisms to obtain a product with better shelf life. © 2017 Society of Chemical Industry.

Response Surface Optimization of Process Parameters and Fuzzy Analysis of Sensory Data of High Pressure-Temperature Treated Pineapple Puree.

The high-pressure processing conditions were optimized for pineapple puree within the domain of 400-600 MPa, 40-60 °C, and 10-20 min using the response surface methodology (RSM). The target was to maximize the inactivation of polyphenoloxidase (PPO) along with a minimal loss in beneficial bromelain (BRM) activity, ascorbic acid (AA) content, antioxidant capacity, and color in the sample. The optimum condition was 600 MPa, 50 °C, and 13 min, having the highest desirability of 0.604, which resulted in 44% PPO and 47% BRM activities. However, 93% antioxidant activity and 85% AA were retained in optimized sample with a total color change (∆E*) value less than 2.5. A 10-fold reduction in PPO activity was obtained at 600 MPa/70 °C/20 min; however, the thermal degradation of nutrients was severe at this condition. Fuzzy mathematical approach confirmed that sensory acceptance of the optimized sample was close to the fresh sample; whereas, the thermally pasteurized sample (treated at 0.1 MPa, 95 °C for 12 min) had the least sensory score as compared to others.

High-Pressure Inactivation of Enzymes: A Review on Its Recent Applications on Fruit Purees and Juices.

In the last 2 decades high-pressure processing (HPP) has established itself as one of the most suitable nonthermal technologies applied to fruit products for the extension of shelf-life. Several oxidative and pectic enzymes are responsible for deterioration in color, flavor, and texture in fruit purees and juices (FP&J). The effect of HPP on the activities of polyphenoloxidase, peroxidase, ß-glucosidase, pectinmethylesterase, polygalacturonase, lipoxygenase, amylase, and hydroperoxide lyase specific to FP&J have been studied by several researchers. In most of the cases, partial inactivation of the target enzymes was possible under the experimental domain, although their pressure sensitivity largely depended on the origin and their microenvironmental condition. The variable sensitivity of different enzymes also reflects on their kinetics. Several empirical models have been established to describe the kinetics of an enzyme specific to a FP&J. The scientific literature in the last decade illustrating the effects of HPP on enzymes in FP&J, enzymatic action on those products, mechanism of enzyme inactivation during high pressure, their inactivation kinetics, and several intrinsic and extrinsic factors influencing the efficacy of HPP is critically reviewed in this article. In addition, process optimization of HPP targeting specific enzymes is of great interest from an industrial approach. This review will give a fair idea about the target enzymes specific to FP&J and the optimum conditions needed to achieve sufficient inactivation during HPP treatment.