Improved stability of phycobiliprotein within liposome stabilized by polyethylene glycol adsorbed cellulose nanocrystals.

This study ascertained the stability of phycobiliprotein (PBP), a bioactive protein from Dulse (Palmaria palmata) loaded within liposomes and stabilized with polyethylene glycol (2000 and 4000 g/mol) and desulfated CNCs (DCs) containing adsorbed polyethylene glycol (DCs-2000 and DCs-4000). The effect of pH, temperature and illumination on the stability of PBP was investigated. Results showed that the temperature had the most significant (p < 0.05) effect on the fluorescence intensity of the PBP, accounting for up to 70% loss of the fluorescence intensity for PBP loaded liposome (PL), PL stabilized with PEG-2000 (PLP-2000) and PEG 4000 (PLP-4000) and PL stabilized with desulfated CNCs (DCs), however, 60% for the PL stabilized with PEG 2000 and PEG 4000 adsorbed CNCs (PLDCs-2000 and PLDCs-4000) at 60 °C compared to those stabilized at 4 °C. A further increase in temperature to 80 °C led to a complete loss of fluorescence. Operating at the extreme pH's of 1.0 and 11.0 resulted in a loss of 90% and 30% fluorescence intensity, respectively for PBP in solution, whereas, 20% and 2% loss was observed for PBP incorporated inside the liposomes. Regarding the effect of illumination, PLDCs-2000 and PLDCs-4000 were the most stable, retaining the fluorescence intensity of PBP up to 70% after 72 h of exposure. This is compared to 85% loss of fluorescence for PBP in solution. Furthermore, at pH of 1.0, there was an increase in average particle size for the PLDCs-2000 and PLDCs-4000 from 189 ± 3 & 206 ± 2 nm to 6464 ± 211 & 6698 ± 317 nm and a charge reversal in the zeta potential from -36 ± 1 & -34 ± 2 to +16 ± 3 & +14 ± 1. Confocal and optical microscopic images confirmed the coalescence of PBP loaded liposome and agglomeration PLDCs-2000 and PLDCs-4000 under acidic pH (<3.0). In contrast, changes in temperature from 4 °C to 100 °C and illumination as a function of time up to 72 h resulted in no change in liposome size and zeta potential.

Comparison of recovery and immunochemical detection of peanut proteins from differentially roasted peanut flour using ELISA.

The effect of heat on extractability and immunoreactivity of proteins from roasted peanut flours and whole peanuts was evaluated using two general protein assays and six commercial peanut ELISA kits, respectively. The highest amount of protein was recovered from roasted peanuts with all ELISAs, while recovery showed a decrease with increasing levels of roasting of the peanut flours. Only the Morinaga kit showed sufficient sensitivity to detect peanut at low concentrations of the dark roast peanut flours. Both the protein and immunoassays indicated a decrease in protein solubility with roasting. The underestimation by immunoassays is a combination of decreased solubility and heat induced changes in the proteins that are being targeted by the ELISA antibodies. These findings suggest that most commercial ELISA kits may not reliably quantify peanut present in dark roast peanut flours at ≤25 ppm.

Structural changes of 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) treated shrimp tropomyosin decrease allergenicity.

The aim of this study was to analyze the effect of AAPH on the conformational structure and allergenicity of shrimp tropomyosin (TM). The structure of AAPH-TM was evaluated by SDS-PAGE, fluorescence, circular dichroism (CD) and ultraviolet light (UV), and the allergenicity was evaluated by in vivo and in vitro methods. Results showed that AAPH can induce structural changes through TM aggregations. These aggregations can decrease the IgG/IgE binding capacity on immunoblot analysis. Further competitive inhibition ELISA (ciELISA) results showed the IC50 of AAPH-TM (AAPH 0-25 mmol/l) changed from 0.086 to 46.2 µg/ml, which correlated with TM structural changes. An RBL-2H3 cell assay showed that release rate of ß-hexosaminidase and histamine decreased by 51.6% and 68.1% with AAPH (5 mmol/l) treatment, respectively. A mouse model showed AAPH-TM could decrease levels of IgE/IgG1, release of histamine and mMCP-1 in sera. In conclusion, AAPH induced TM aggregation can cause structural changes and decrease the allergenicity.

Evaluation of the efficiency of three extraction conditions for the immunochemical detection of allergenic soy proteins in different food matrices.

BACKGROUND: Recent studies have shown the need to improve soy allergen extraction using different extraction conditions to ensure more accurate results in allergen detection. This study investigated some of these extraction conditions to confirm that these methods, especially ultrasound-assisted extraction (UAE) and the use of Laemmli buffer instead of the conventional extraction with phosphate-buffered saline (PBS), could be helpful in improving the extraction step in allergen detection. RESULTS: Higher total soluble protein was obtained in all samples extracted with Laemmli buffer alone and in combination with ultrasound. For immunochemical detection of soy proteins by enzyme-linked immunosorbent assay (ELISA), comparable detection was observed in extracts from all extraction conditions in all commercial samples with the exception of table cracker and veggie burger, where significantly higher detection was seen in extracts from Laemmli buffer only. For the dry mix and cookie samples, the degree of soy protein detection with ELISA varied among the different extraction conditions, but overall, extraction with only Laemmli buffer showed higher detection. CONCLUSION: Laemmli buffer with conventional extraction and UAE may be better alternatives or additional extraction methods in soy allergen detection. Different food matrices performed differently (whether it was for the recovery of total proteins or detection by ELISA) under different extraction conditions. © 2017 Society of Chemical Industry.

Shrimp tropomyosin retains antibody reactivity after exposure to acidic condition.

BACKGROUND: Although shrimp can be found in certain high acid food matrices, the allergenic capacity of shrimp tropomyosin exposed to low pH condition has not been fully clarified. Thus, a model marinade comprising white vinegar adjusted to different pH was used to determine the effects of acid-induced denaturation on the immunoreactivity of tropomyosin. RESULTS: Whole shrimp experienced either swelling or shrinkage after marination depending on the vinegar pH and the final muscle pH. The extractability of soluble myofibrillar proteins was reduced significantly among shrimp marinated in vinegar at pH 1.0-3.5, and a substantial amount of tropomyosin was retained in the insoluble pellets. Consequently, the immunoglobulin E (IgE)-binding capacity of tropomyosin was significantly lower in the soluble protein fraction of shrimp marinated at pH 1.0-3.5 compared with samples marinated at pH 4.8 and control. However, tropomyosin in the insoluble protein fraction of all marinated shrimp showed strong IgE-binding capacity at all marinating conditions. CONCLUSION: Thus, tropomyosin in shrimp exposed to low pH condition retained its allergenic capacity owing to the conservation of its linear epitopes. Analysis of the insoluble protein fraction was crucial for the accurate determination of the effect of low pH condition on the immunoreactivity of this allergen. © 2017 Society of Chemical Industry.

Effects of Microwave and Ultrasound Assisted Extraction on the Recovery of Soy Proteins for Soy Allergen Detection.

The extraction of soy proteins for soy allergen detections is conventionally achieved with PBS buffer for at least 2 h at room temperature or 4 °C. This method has been reported to be inefficient due to time consumption and inadequate protein extraction resulting in false negative allergen detection and mislabeling of foods containing allergenic proteins. This study investigated the application of microwave (MAE) and ultrasound assisted extraction (UAE) techniques to extract and improve recovery of allergens from various soy matrices. Soy proteins were extracted from raw soy flour, soy protein isolate (SPI) and soy milk using MAE at 60, 70, and 100 °C for 5 and 10 min and UAE at 4 and 23 °C for extraction times of 1, 5, and 10 min with PBS, Laemmli and urea buffers. Extracts were analyzed for total proteins, protein profile, and antibody-based detection (ELISA) of soy proteins. Conventional extraction with each of the buffers was used as controls. Overall, proteins recovered from MAE and UAE samples were higher than recoveries from the controls in all soy matrices. Under all extraction conditions, Laemmli and urea buffer recovered more proteins than PBS. Electrophoresis analysis of protein showed bands around 75, 50, and 33 kDa indicating the presence of soy allergenic proteins ß-conglycinin and glycinin, in all samples. Using sandwich ELISA, control and UAE extracts resulted in high soy protein detection but this reduced in MAE extracts.

Effects of buffer additives and thermal processing methods on the solubility of shrimp (Penaeus monodon) proteins and the immunoreactivity of its major allergen.

This study examines the potential of two buffer additives (Tween 20 and DTT) to improve the solubility of proteins from shrimp subjected to different heat treatments and the allergenicity of tropomyosin in the extracts. The concentration of soluble proteins extracted by all the buffers from processed shrimp was significantly reduced compared with untreated samples. The concentration of total soluble proteins from heat treated shrimp increased significantly when phosphate buffer containing both surfactant and reducing agent was used as the extraction buffer. However, the concentrations of heat-stable proteins in the buffers were mostly similar. The electrophoretic profile of extracted proteins showed that tropomyosin is very stable under the different heat treatment methods used in this study except for high pressure steaming where the intensity of tropomyosin band was reduced. Competitive inhibition ELISA showed that high pressure steaming reduced the allergenicity of tropomyosin compared with other heat treatments methods.

Comparison of microwave, ultrasound and accelerated-assisted solvent extraction for recovery of polyphenols from Citrus sinensis peels.

Peel of Citrus sinensis contains significant amounts of bioactive polyphenols that could be used as ingredients for a number of value-added products with health benefits. Extraction of polyphenols from the peels was performed using a microwave-assisted extraction (MAE) technique. The effects of aqueous acetone concentration, microwave power, extraction time and solvent-to-solid ratio on the total phenolic content (TPC), total antioxidant activity (TAA) (using DPPH and ORAC-values) and individual phenolic acids (IPA) were investigated using a response surface method. The TPC, TAA and IPA of peel extracts using MAE was compared with conventional, ultrasound-assisted and accelerated solvent extraction. The maximum predicted TPC under the optimal MAE conditions (51% acetone concentration in water (v/v), 500 W microwave power, 122 s extraction time and 25 mL g(-1) solvent to solid ratio), was 12.20 mg GAE g(-1) DW. The TPC and TAA in MAE extracts were higher than the other three extracts.

Optimization of microwave-assisted extraction of polyphenols from Myrtus communis L. leaves.

Phytochemicals, such as phenolic compounds, are of great interest due to their health-benefitting antioxidant properties and possible protection against inflammation, cardiovascular diseases and certain types of cancer. Maximum retention of these phytochemicals during extraction requires optimised process parameter conditions. A microwave-assisted extraction (MAE) method was investigated for extraction of total phenolics from Myrtus communis leaves. The total phenolic capacity (TPC) of leaf extracts at optimised MAE conditions was compared with ultrasound-assisted extraction (UAE) and conventional solvent extraction (CSE). The influence of extraction parameters including ethanol concentration, microwave power, irradiation time and solvent-to-solid ratio on the extraction of TPC was modeled by using a second-order regression equation. The optimal MAE conditions were 42% ethanol concentration, 500 W microwave power, 62 s irradiation time and 32 mL/g solvent to material ratio. Ethanol concentration and liquid-to-solid ratio were the significant parameters for the extraction process (p<0.01). Under the MAE optimised conditions, the recovery of TPC was 162.49 ± 16.95 mg gallic acidequivalent/gdry weight(DW), approximating the predicted content (166.13 mg GAE/g DW). When bioactive phytochemicals extracted from Myrtus leaves using MAE compared with UAE and CSE, it was also observed that tannins (32.65 ± 0.01 mg/g), total flavonoids (5.02 ± 0.05 mg QE/g) and antioxidant activities (38.20 ± 1.08 µg GAE/mL) in MAE extracts were higher than the other two extracts. These findings further illustrate that extraction of bioactive phytochemicals from plant materials using MAE method consumes less extraction solvent and saves time.

Effect of processing on phenolic antioxidants of fruits, vegetables, and grains--a review.

Understanding the influence of processing operations such as drying/dehydration, canning, extrusion, high hydrostatic pressure, pulsed electric field, and ohmic heating on the phytochemicals of fruits, vegetables, and grains is important in retaining the health benefiting properties of these antioxidative compounds in processed food products. Most of the previous investigations in the literature on the antioxidants of fruits, vegetables, and grains have shown that food-processing operations reduced the antioxidants of the processed foods, which is also the usual consumer perception. However, in the last decade some articles in the literature reported that the evaluation of nutritional quality of processed fruits and vegetables not only depend on the quantity of vitamin C but should include analyses of other antioxidant phytochemicals and antioxidant activity. Thermal processing increased the total antioxidant activity of tomato and sweet corn. Most importantly, analysis also depends on the condition, type, and mechanism of antioxidant assays used. This review aims to provide concise information on the influence of various thermal and nonthermal food-processing operations on the stability and kinetics of health beneficial phenolic antioxidants of fruits, vegetables, and grains.

Thermal degradation of anthocyanins from purple potato (cv. Purple Majesty) and impact on antioxidant capacity.

Degradation parameters of purified anthocyanins from purple-fleshed potato (cv. Purple Majesty) heated at high temperatures (100-150 °C) were determined. Purified anthocyanins, prepared by removing salts, sugars, and colorless nonanthocyanin phenolics from the crude extract, were monitored and quantified using HPLC and spectrophotometry for heat-induced degradation products. Separation of colorless phenolics from the anthocyanins was confirmed using HPLC at two wavelengths, 280 and 520 nm. The degradation kinetics of purified anthocyanins followed a first-order reaction with reaction rate constants (k values) of 0.0262-0.2855 min(-1), an activation energy of 72.89 kJ/mol, thermal death times (D values) of 8.06-8789 min, and a z value of 47.84 °C over the temperature range of 100-150 °C. The enthalpy and entropy of activation were 59.97 kJ/mol and -116.46 J/mol·K, respectively. The antioxidant capacity in the purified anthocyanins, measured by DPPH and ABTS assays, was increased after the thermal treatment, indicating antioxidant activities of degradation products in the samples.

Bioactivity of antioxidants in extruded products prepared from purple potato and dry pea flours.

Measuring antioxidant activity using a biologically relevant assay adds important evidence to aid in understanding the role of phytochemicals based on data from in vivo and chemical assays of extrusion processed purple potato and pea flours. A cellular antioxidant activity assay could provide biologically relevant information on bioactive compounds in raw as well as processed food products. The objective of this study was to investigate the complete phytochemical profiles, antioxidant activity, cellular antioxidant activity, and their contribution to bioactivity in purple potato flour, dry pea flour, raw formulations, and extrusion cooked products prepared with the above ingredients. The free fraction of extracts contributed 68, 64, and 88% to total phenolics, total antioxidant activity (ORAC value), and total flavonoids, respectively, in purple potato flour (PPF). Similarly, extracts in the free fraction contributed 87, 86, and 64% to total phenolics, total antioxidant activity (ORAC value), and total flavonoids, respectively, in dry pea flour (DPF). The amount of total phenolics and total flavonoids in purple potato flour and the antioxidant activity of PPF and DPF were comparable to published data. However, a higher amount in the total flavonoids and lower in the total phenolics of DPF were observed. Caffeic, p-coumaric, and ferulic acids were mostly observed in the bound extracts of raw formulations as in the extrudates, whereas chlorogenic acid was predominant in the free extracts. The extruded products had significantly higher (p < 0.05) content of total phenolics, ORAC antioxidant activity, and flavonoids, compared to the raw formulations. Extrusion processing increased the cellular antioxidant activity of the extrudates prepared from 35:65 and 50:50 PPF/DPF (w/w) of ingredients compared with control raw formulations in a dose-dependent manner. Increase of PPF significantly increased (p < 0.05) the cellular antioxidant activity of 35-50% PPF formulations.

Effect of extrusion on the antioxidant capacity and color attributes of expanded extrudates prepared from purple potato and yellow pea flour mixes.

Foods with antioxidant capacity provide protection against cardio-vascular, certain forms of cancers, and Alzheimer's diseases caused by oxidative damages and contribute health benefits. The effect of extrusion cooking on the antioxidant capacity and color attributes of extruded products prepared from 3 selected formulations of purple potato and yellow pea flours using a co-rotating twin screw extruder were studied. Expansion ratios of the extruded products varied from 3.93 to 4.75. The total antioxidant capacities (TAC) of the extruded products, using DPPH assay, were 3769 to 4116 µg trolox equivalent/g dry weight sample and not significantly different (P > 0.05) from their respective raw formulations. The total phenolic contents (TP) of the extruded products varied from 2088 to 3766 µg of gallic acid equivalent/g dry weight sample and retained 73% to 83% of the TP from the raw formulations after extrusion. The total anthocyanins contents (TA) in the extrudates were 0.116 to 0.228 mg of malvidin-3-glucosides/g dry weight sample. Compared with their raw formulations, significant losses (60% to 70%) of the TA in the extruded products occurred due to extrusion cooking. Browning indices and color attributes such as brightness, chroma, and hue angle agreed with degradation of anthocyanins in the extruded products. However, extrusion cooking retained antioxidant capacities of the raw formulations in the extruded products either in their natural forms or degraded products with radical scavenging activity. This study demonstrated the potential for the production of puffed extruded food products with the improved antioxidant content from colored potatoes and pulse formulations.