Efficacy of sodium chlorite as an inhibitor of enzymatic browning in apple slices

Sodium chlorite (SC) is an effective sanitizer for inhibiting microbial growth. This investigation was conducted to determine the efficacy of SC as a browning control agent for use on fresh-cut apple slices, applied alone, or in conjunction with organic acids. Additionally, the authors compared the efficacy of SC to that of acidified sodium chlorite (ASC) and to several other salts and examined the effect of pH and several different organic acids on efficacy of SC. The fresh-cut apple slices were dipped in treatment solutions for 1 min, then drained and placed in plastic containers at 20 °C for 24 h, and finally stored in polyethylene bags at 5 °C for 2 weeks. Color was measured periodically during storage. Lightness (L) values for all treated and control samples measured at 4 h, 24 h, and 2 weeks of storage were compared to L value for untreated samples measured immediately after cutting. Percent decrease in L-values was calculated for each sample at each time interval. Apple slices treated in ASC or SC solution had a significantly smaller decrease in L value indicating less browning than those treated in citric acid or water control at 4 h (P < 0.01), and with the exception of 1 g L⁻¹ ASC and 0.1 g L⁻¹ SC, all other ASC and SC treated slices still had significantly less browning than those for the water control (P < 0.01) at 24 h. After 2 weeks of storage, only SC (0.5–1.0 g L⁻¹), sodium bisulfite (0.5 g L⁻¹) and calcium l-ascorbate (10 g L⁻¹) continued to inhibit browning. Treatment with 0.5 g L⁻¹ SC and pH adjusted in the range from 3.9 to 6.2 using citric acid (CA) reduced browning more effectively than 0.5 g L⁻¹ SC without pH adjustment. Two organic acids, salicylic acid and cinnamic acid, when added to SC solution, were found to achieve even better inhibition of browning than CA at the same pH value.     

Mechanism for the inhibition of apple juice enzymatic browning by Palo Fierro (desert ironweed) honey extract and other natural compounds

Inhibition of Red Delicious apple juice browning and apple polyphenoloxidase (PPO) activity by four natural anti-browning agents [palo fierro honey extract (PFH), caffeic acid phenethyl ester (CAPE), l-cysteine (Cys), and 4-hexylresorcinol (4HR)] has been studied. Browning of apple juice was markedly reduced by 4HR, Cys, and PFH, however, little effect was observed with CAPE. PFH, CAPE, and 4HR showed high inhibition of apple juice PPO. When the inhibition mechanism of these compounds over partially purified apple PPO was studied using chlorogenic acid as substrate, PFH acted as a noncompetitive inhibitor of apple PPO, with K_i = αK = 0.8 g/L, and IC₅₀ ranging 1.1-1.7 g/L. Cys showed an inhibitory pattern which could not be fitted to any type of reversible inhibition (IC₅₀ 0.02-0.03 g/L), while CAPE showed low inhibitory activity, which could be due to co-precipitation processes. PFH could be considered as an effective natural compound to reduce apple juice browning.     

Biochemical properties and potential endogenous substrates of polyphenoloxidase from chufa (Eleocharis tuberosa) corms

Polyphenoloxidase (PPO) was partially purified from chufa corms through ammonium sulphate precipitation and dialysis. Biochemical properties of chufa PPO were analysed using exogenous substrate catechol. Optimal pH and temperature for PPO activity were 5 and 45 °C. Ethylenediaminetetraacetic acid disodium salt and l-cysteine could not inhibit the PPO activity. However, sodium thiosulphate pentahydrate exhibited the strongest inhibiting effect, followed by ascorbic acid and anhydrous sodium sulphite. Except for K⁺, other metal ions such as Zn²⁺, Cu²⁺, Fe³⁺, Ca²⁺, Fe²⁺ and Na⁺ accelerated the enzymatic reaction between catechol and PPO. Kinetic analysis showed that the apparent K_m and V_max values were around 10.77 mM and 82 units/ml min. In addition, (−)-gallocatechin gallate, (−)-epicatechin gallate and (+)-catechin gallate isolated and identified from chufa corms were supposed to be the potential endogenous PPO substrates due to their ortho-diphenolic or pyrogallolic structures. These polyphenols might be catalysed by PPO, resulting in the browning of chufa corms after fresh-cut processing.     

Probiotic beverage from cashew apple juice fermented with Lactobacillus casei

This study optimized the conditions of Lactobacillus casei NRRL B-442 cultivation in cashew apple juice, as well as, determined the proper inoculum amount and fermentation time. Moreover, it was investigated the survivability ability of L. casei in cashew apple juice during refrigerated storage (4 °C) for 42 days. The optimum conditions for probiotic cashew apple juice production were initial pH 6.4, fermentation temperature of 30 °C, inoculation level of 7.48 Log CFU/mL (L. casei) and 16 h of fermentation process. It was observed that the L. casei grew during the refrigerated storage. Viable cell counts were higher than 8.00 Log CFU/mL throughout the storage period (42 days). The values of lightness, yellowness and total color change increased and the values of redness reduced along the fermentation and refrigerated storage periods. The fermented juice with L. casei is a good and healthy alternative functional food containing probiotics. Cashew apple juice showed to be as efficient as dairy products for L. casei growth.     

Dual effectiveness of sodium chlorite for enzymatic browning inhibition and microbial inactivation on fresh-cut apples

The dual effectiveness of sodium chlorite for browning inhibition and microbial inactivation on fresh-cut apples was investigated and compared to other anti-browning and antimicrobial agents. Results indicate that sodium chlorite significantly (P < 0.001) inhibited the browning reaction of fresh-cut Red Delicious apples stored at 5 °C for 14 days. This treatment also significantly reduced polyphenol oxidase activities. Treatments with acidified sodium chlorite, calcium chloride, or calcium ascorbate exhibited strong inhibition on apple browning during the early storage, these treatment effects diminished after 7 days in storage. Combining calcium chloride with sodium chlorite further significantly (P < 0.001) improved the firmness of apple slices, and browning inhibition during storage. Additionally, treatments with acidified sodium chlorite, sodium chlorite, or the combination of sodium chlorite and calcium chloride significantly (P < 0.001) reduced Escherichia coli populations on fresh-cut apples by 3.0, 3.6, and 3.9 log cfu g⁻¹ over the water control. The dual effectiveness of sodium chlorite to inhibit enzymatic browning and inactivate E. coli may allow this compound to achieve a prominent role in improving the quality and safety of products in the fresh-cut apple and other food industries.     

Prebiotic effect of fermented cashew apple (Anacardium occidentale L) juice

Leuconostc mesenteroides B-512F and L. mesenteroides B-742 were cultivated in clarified cashew apple juice to produce prebiotic oligosaccharides. Yeast extract (20 g/L); K₂HPO₄ (g/L) and sucrose (50 g/L) were added to the juice to promote the microbial growth and dextransucrase production. Initial pH was adjusted to 6.5 with H₃PO₄. Fermentations were carried out at 30 °C and 150 rpm for 24 h. The prebiotic effect of the fermented cashew apple juice, containing oligosaccharides, was evaluated through the Lactobacillus johnsonii B-2178 growth. L. johnsonii was incubated for 48 h using fermented cashew apple juice as substrate. Lactobacillus growth was compared to the microbial growth in non-fermented juice and in MRS broth. L. johnsonii growth in the fermented cashew apple juice was threefolds the observed growth in the non-fermented juice.     

Aseptically packaged UHPH-treated apple juice: Safety and quality parameters during storage

Ultra-high pressure homogenization (UHPH) at 300 MPa and 4 °C inlet temperature were used to preserve apple juice, and shelf-life evaluation of aseptically packaged juice was investigated. After processing Tetra Brik containers were stored at temperatures of 4, 10, 20 and 30 °C during 60 days. In this article, the effect of processing on the spoilage inactivation was evaluated after processing and during the storage trial. Non-germinated and germinated spores were found in the UHPH-treated juice, being an inactive population during storage. Patulin content was also not modified by UHPH processing, but a significant decrease was observed during storage at 30 °C (P < 0.05). Polyphenoloxidase (PPO) and pectinmethylesterase (PME) activity was not found after UHPH-processing and during storage.A kinetic study of post-processing quality loss was conducted. Vitamin C, chlorogenic acid, total polyphenols and color change were measured during storage study and were used to model the UHPH-treated apple juice shelf-life. Loss of vitamin C was correlated with the hydroxymethylfurfural (HMF) accumulation (0.59, P < 0.05). A limiting quality parameter was polyphenolic content. UHPH-treated apple juice stored at 4 °C was found to show a shelf-life for about 21 months by preserving the color characteristics of the juice with low HMF accumulation. From 15 °C changes in quality parameters were more evident.     

Kinetic model of apple juice enzymatic browning in the presence of cyclodextrins: The use of maltosyl-β-cyclodextrin as secondary antioxidant

Enzymatic browning reactions limit the commercial shelf life of apple juice, so that colour preservation during storage is one of the main objectives of fruit processors. In this paper, the colour of fresh apple juice was evaluated in the presence of different types of cyclodextrins (CDs) (α-CD, β-CD, γ-CD and maltosyl-β-CD), compounds that bind or complex polyphenol oxidase substrates. The effectiveness of CDs as browning inhibitors was determined as the difference between the colours observed in the CD-treated sample and the controls, using the colour space CIE-L^∗, a^∗, b^∗ system. Although the effect of CDs on apple juice enzymatic browning has been studied, the action mechanism involved remains a subject of controversy. In this work, we have kinetically modelled apple juice enzymatic browning in the absence and presence of maltosyl-β-CD. The complexation constant between the mixtures of diphenols present in apple juice and maltosyl-β-CD was calculated (K_c = 4.09 mM⁻¹). Different concentrations of maltosyl-β-CD modified the evolution of lightness (L^∗), total colour (ΔE^∗) and hue angle (H^∗) because of the higher affinity constant it shows for the compounds responsible for apple juice browning than do α-CD, β-CD and γ-CD. Moreover, in this paper we show that maltosyl-β-CD can enhance the ability of ascorbic acid to prevent the enzymatic browning due to the protective effect of maltosyl-β-CD against ascorbic acid oxidation. Hence, maltosyl-β-CD seems to act as a “secondary antioxidant”, reducing apple juice browning and enhancing the naturally occurring antioxidant capacity of a food.     

Properties of polyphenol oxidase from Anamur banana (Musa cavendishii)

Polyphenol oxidase (PPO) was extracted from Anamur banana, grown in Turkey, and its characteristics were studied. The optimum temperature for banana PPO activity was found to be 30 °C. The pH-activity optimum was 7.0. From the thermal inactivation studies, in the range 60–75 °C, the half-life values of the enzyme ranged from 7.3 to 85.6 min. The activation energy (E_a) and Z values were calculated to be 155 kJ mol⁻¹ and 14.2 °C, respectively. K_m and V_max values were 8.5 mM and 0.754 OD₄₁₀ min⁻¹, respectively. Of the inhibitors tested, ascorbic acid and sodium metabisulphite were the most effective.     

Changes in bioactive compounds and antioxidant capacity of fresh-cut cashew apple

In this study, ascorbic acid, total polyphenols and proanthocyanidins of fresh-cut cashew apple were quantified. Antioxidant capacity was determined in whole juice and in polyphenols extracts by three methods: 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) and ??-carotene bleaching. Effect of cutting and storage for 24 h at 2 °C, 27 °C and 40 °C on these compounds were also evaluated. Cashew apple presented 163 mg of ascorbic acid per 100 g of fresh weight (FW). Soluble and hydrolysable polyphenols contents were 12.79 mg GAE/100 g FW and 18.53 mg GAE/100 g FW and proanthocyanidins were 9.27 mg/100 g FW. Antioxidant capacity of juice and polyphenols extract was high for DPPH method. Storage temperatures affected bioactive compounds on cut cashew apple. The content of ascorbic acid decreased in all temperatures. Proanthocyanidins were more sensitive to 40 °C than to other temperatures. The content of polyphenols and antioxidant capacity of juice by DPPH assay did not change. However, the reducing power was lower in samples kept at high temperatures. A strong positive correlation between ascorbic acid and FRAP (r = 0.99) and a negative correlation between DPPH and FRAP (r = − 0.79) were observed. No correlations were found between polyphenols and antioxidant capacity indicating the importance of phenolic composition in the extracts. The results confirm the importance of temperature and injury on the quality of cashew apple.     

Effects of aqueous chlorine dioxide treatment on browning of fresh-cut lotus root

Effect of aqueous chlorine dioxide (ClO₂) treatment on browning of fresh-cut lotus root (FLR) was investigated to explore the feasibility to apply ClO₂ for browning inhibition of fresh-cut products. Cut lotus roots were treated in ClO₂ solutions at different concentrations (10, 50 and 100 mg/l) for different time (5, 10 and 15 min), followed by chilled storage for 8-10 days at 4 °C. Color parameters (L^∗, a^∗ and b^∗), polyphenol oxidase (PPO) activity and overall visual quality (OVQ) were measured at one-day interval during storage. Results showed that higher ClO₂ concentration and longer treatment time can provide better inhibitory effects on the browning of FLR. ClO₂ concentration, treatment time and storage time were three significant factors (P < 0.05) and some significant interactions were observed. PPO activities were largely inhibited by 100 mg/l ClO₂ treatment for 10 min. The 100 mg/l ClO₂ treatment maintained high OVQ scores during 10-day storage; while 50 mg/l ClO₂ treatment was acceptable for maintaining OVQ during 4-day storage. ClO₂ treatment was demonstrated to be a promising alternative approach to control browning and improve OVQ of FLR.     

Change of polyphenol oxidase activity, color, and browning degree during storage of cloudy apple juice treated by supercritical carbon dioxide

The effects of supercritical carbon dioxide (SCCO₂) treatment of 8, 15, 22, and 30 MPa for 60 min at 55 °C on polyphenol oxidase (PPO) activity, color, and browning degree in cloudy apple juice during storage at 4 °C for 4-weeks were investigated. The SCCO₂ treatment had significant effects on inactivation of PPO and the least residual activity of PPO was 38.50% at 30 MPa. The restoration of PPO residual activity after SCCO₂ treatment was also observed, which was dependent on the pressure level. A greater reduction of lightness L and a minor increase of redness a of cloudy apple juices after SCCO₂ treatment occurred. Moreover, the total color difference (ΔE), which was significantly less than that of untreated sample, was decreased by enhancing the pressure level. The changes of lightness L and browning degree A during storage were well fitted to a first-order kinetic model. The rate constants of k _L and k _A of samples subjected to SCCO₂ treatment reduced from 4.75×10⁻² to 0.42×10⁻² and 37.19×10⁻² to 8.02×10⁻², respectively, when pressure increased from 0 MPa (untreated sample) to 30 MPa.     

Quantification and characterisation of polyphenol oxidase from vanilla bean

Polyphenol oxidase (PPO) of Vanilla planifolia Andrews beans was extracted and purified through ammonium sulphate precipitation, dialysis, and gel filtration chromatography. PPO activity was measured by improved UV technique using 4-methylcatechol and catechol as substrates increasing substantial sensitivity of previous procedure. The optimum pH and temperature for PPO activity were found to be 3.0 and 3.4 and 37 °C, respectively. K_m and V_max values were found to be 10.6 mM/L and 13.9 OD₃₀₀ min⁻¹ for 4-methylcatechol and 85 mM/L and 107.2 OD₃₀₀ min⁻¹ for catechol. In an inhibition test, the most potent inhibitor was found to be 4-hexylresorcinol followed by ascorbic acid. The thermal inactivation curve was biphasic. Activation energy (E_a) and z values were calculated as 92.10 kJ mol⁻¹ and 21 °C, respectively.     

Characterization of polyphenol oxidase from Napoleon grape

Polyphenol oxidase (PPO) from Napoleon grape was isolated using a two-phase partitioning approach with Triton X-114. The enzyme was purified in a latent form and could be optimally activated by the presence of 0.2% of sodium dodecyl sulphate (SDS) at pH 6.0. In the absence of SDS, the enzyme showed maximum activity at acid pH (3.0). The enzyme was kinetically characterized at pH 3.0 and pH 6.0 in the presence of 0.2% of SDS, using 4-tert-butylcatechol (TBC) as a substrate. The V_m/K_M ratio showed that Napoleon grape PPO presents greater affinity for TBC at acid pH (0.1 min⁻¹) that at pH 6.0 in the presence of SDS (0.02 min⁻¹). The enzyme was highly heat stable, 80% of activity remaining at 70 °C. Selected inhibitors were also studied, tropolone being the most active with a K_i value of 27 μM at acid pH and pH 6.0 in the presence of 0.2% SDS.     

Using polysaccharide-based edible coatings to maintain quality of fresh-cut Fuji apples

The effect of alginate and gellan-based edible coatings on the shelf-life of fresh-cut Fuji apples packed in trays with a plastic film of a known permeability to oxygen (110 cm³ O₂ m⁻² bar⁻¹ day⁻¹) was investigated by measuring changes in headspace atmosphere, color, firmness and microbial growth during 23 days of storage at 4 °C. Concentration of O₂ and CO₂ in the package was measured and no significant differences between coated and uncoated fresh-cut apples were observed. Ethylene concentration in coated apples seemed to be delayed since it remained below 50 μl l⁻¹ throughout the whole refrigerated storage period, while production of this gas was detected in uncoated apples from the very initial days of storage. Coated apple wedges exhibited ethanol and acetaldehyde formation from the second week of storage indicating fermentative metabolism. Polymers were crosslinked with a calcium chloride solution, to which the antibrowning agent N-acetylcysteine was added, being incorporated into the coatings formulation and helping to maintain firmness and color of apple wedges during the entire storage time. The application of the edible coatings also retarded the microbiological deterioration of fresh-cut apples. Alginate and gellan edible coatings effectively prolonged the shelf-life of Fuji apple wedges by 2 weeks of storage compared with the control apple slices which showed a considerable cut surface browning and tissue softening from the very early days of storage, limiting their shelf-life to less than 4 days.     

Kinetic characterisation and thermal inactivation study of polyphenol oxidase and peroxidase from table grape (Crimson Seedless)

Polyphenol oxidase (PPO) and peroxidase (POD) were extracted from a table grape (Crimson Seedless) using Triton X-114 and characterized using spectrophotometric methods. Both PPO and POD were activated by acid shock. However, in the presence of the anionic detergent sodium dodecil sulphate (SDS), PPO was activated whereas POD was inactivated. The enzymes were kinetically characterized and both followed Michaelis–Menten kinetics, although with different values of their kinetic parameters. The V_m/K_m ratio showed that Crimson Seedless grape PPO presents a similar affinity for 4-tert-butyl-catechol (TBC) whether activated by acid shock (0.018 min⁻¹) or SDS (0.023 min⁻¹). With regards to POD, the K_m and V_m values for 2,2′-azinobis(3-ethylbenzothiazolinesulphonic acid) (ABTS) were 0.79 mM and 1.20 μM/min, respectively. In the case of H₂O₂, the K_m and V_m value were 0.4 mM and 0.93 μM/min, respectively. PPO and POD showed similar thermostability, losing >90% of relative activity after only 5 min of incubation at 78 °C and 75 °C, respectively. In addition, PPO´s activation energy was similar to that obtained for POD (295.5 kJ/mol and 271.9 kJ/mol, respectively).     

Physicochemical changes in fresh-cut wax apple (Syzygium samarangenese [Blume] Merrill & L.M. Perry) during storage

Physicochemical changes, such as peel and flesh colours, total anthocyanin content, browning index, firmness, total soluble solid (TSS), titratable acidity (TA), sugar acid ratio (TSS/TA), antioxidant capacity, total phenolic content and ascorbic acid content, in fresh-cut Taaptimjan wax apple fruit stored at 4 ± 2 °C and 12 ± 2 °C for 7 days were investigated. The skin of fresh-cut fruit stored at 4 ± 2 °C showed higher a^∗ value, chroma and total anthocyanin content and lower hue angle than those stored at 12 ± 2 °C. Lightness (L^∗ value) and whiteness index of the fresh-cut fruit flesh stored at 12 ± 2 °C showed significantly lower than those stored at 4 ± 2 °C which related to an significant increase in browning index. Firmness, total soluble solid, titratable acidity and sugar acid ratio did not significant changes during storage. Antioxidant capacity and total phenolic content increased throughout storage. Ascorbic acid content of the fresh-cut fruit stored at 4 ± 2 °C remained constant throughout storage whilst ascorbic content at 12 ± 2 °C decreased and was lower than that at 4 ± 2 °C. At 4 ± 2 °C antioxidant capacity and ascorbic acid content were higher than that stored at 12 ± 2 °C whilst there was no significant difference in total phenolic content. In conclusion, the reduction of whiteness index and the increase in browning index of fresh-cut wax apple flesh were the key factors affecting its quality and storage at 4 ± 2 °C could reduce the change in the flesh colour and maintained the peel colour and nutritional values of fresh-cut wax apple fruit during storage.     

Effect of temperature and air velocity on drying kinetics, antioxidant capacity, total phenolic content, colour, texture and microstructure of apple (var. Granny Smith) slices

The aim of this work was to study the effect of temperature and air velocity on the drying kinetics and quality attributes of apple (var. Granny Smith) slices during drying. Experiments were conducted at 40, 60 and 80 °C, as well as at air velocities of 0.5, 1.0 and 1.5 m s⁻¹. Effective moisture diffusivity increased with temperature and air velocity, reaching a value of 15.30 × 10⁻⁹ m² s⁻¹ at maximum temperature and air velocity under study. The rehydration ratio changed with varying both air velocity and temperature indicating tissue damage due to processing. The colour difference, ΔE, showed the best results at 80 °C. The DPPH-radical scavenging activity at 40 °C and 0.5 m s⁻¹ showed the highest antioxidant activity, closest to that of the fresh sample. Although ΔE decreased with temperature, antioxidant activity barely varied and even increased at high air velocities, revealing an antioxidant capacity of the browning products. The total phenolics decreased with temperature, but at high air velocity retardation of thermal degradation was observed. Firmness was also determined and explained using glass transition concept and microstructure analysis.     

Purification and characterization of polyphenol oxidase from Ferula sp.

Polyphenol oxidase (PPO) of several Ferula sp. was extracted and purified through (NH₄)₂SO₄ precipitation, dialysis, and gel filtration chromatography. Leaf and stem extracts were used for the determination of enzyme properties. Optimum conditions, for pH, temperature, and ionic strength were determined. The best substrates of PPO were catechol for leaf and (−) epicatechin for stem samples. Optimum pH and temperature were determined. K_M and V_max values were 2.34 × 10⁻³ M and 8541 EU/ml for catechol, and 2.89 × 10⁻³ M and 5308 EU/ml for (−) epicatechin. The most effective inhibitor was sodium diethyl dithiocarbamate for leaf samples and sodium metabisulphite for stem samples. Both inhibitors indicated competitive reactions. PPO showed irreversible denaturation after 40 min at 60 °C.     

Non-enzymatic browning in clarified cashew apple juice during thermal treatment: Kinetics and process control

The effect of thermal treatment on clarified cashew apple juice was studied at temperatures from 88 to 121 °C. Changes in colour measured with colorimetric parameters (reflectance spectra, colour difference and CIELAB), and the variation in ascorbic acid, 5-hydroxymethylfurfural (5-HMF) and sugar content were used to evaluate non-enzymatic browning. Kinetic models were applied to the changes in reflectance spectra, ascorbic acid and 5-HMF. The effect of temperature on kinetic constants was described by an Arrhenius type equation. The sugar content remained constant during thermal treatment and did not affect non-enzymatic browning, which was mainly affected by degradation of ascorbic acid. The kinetic models were used to optimise and control the thermal treatment for clarified cashew apple juice.