Identification and analysis of cell wall glycan epitopes and polyphenol oxidase in pawpaw (Asimina triloba [L.] Dunal) fruit pulp as affected by high pressure processing and refrigerated storage.

This research explores the cell wall composition and polyphenol oxidase activity of two pawpaw (Asimina triloba) fruit varieties, Susquehanna and Green River Belle, that were subjected to high pressure processing and 45 days of refrigerated storage. We hypothesize that high pressure processing may inhibit enzymatic action responsible for pawpaw's deleterious postharvest tissue softening and browning. Glycome profiling uses mAb groupings that recognize 19 groups of glycan epitopes present in most major classes of cell wall glycans and was used to determine cell wall composition. Results show that both varieties have typical type I primary cell walls of flowering dicots. However, differences in the fine cell wall structure between the varieties can be inferred and the varieties behaved differently during refrigerated storage, likely indicating of a difference in cell wall-modifying enzymes present in the primary cell walls. High pressure processing treatment does not seem to be effective at eliminating polyphenol oxidase activity.

Valorization of underutilized North American pawpaw (Asimina triloba): investigation as a lipid oxidation inhibitor in turkey homogenate model system.

BACKGROUND: The objective of this study was to characterize the ability of extracts from nine varieties of pawpaw pulp standardized to the phenolics level of 0.1% grape seed extract (GSE) on inhibition of the formation of thiobarbituric reactive substances (TBARS) in a turkey model system. The antioxidant activity of the extracts was also determined using four common assays. RESULTS: Over the 240 min sampling time, the standardized pawpaw extracts from all nine varieties were as effective as GSE in inhibiting TBARS formation in turkey muscle homogenate compared to the untreated control. Extracts from all pawpaw varieties and GSE began to inhibit TBARS formation at 60 min of incubation, and by 240 min TBARS were reduced from 35 µmol malondialdehyde kg-1 tissue in the homogenate to which no antioxidant was added to 4-18 µmol malondialdehyde kg-1 tissue in the antioxidant-enriched extracts. There does not appear to be a clear relationship between inhibition of TBARS and any of the antioxidant capacity measurements (ORAC, DPPH inhibition, reducing potential as measured by FRAP assay, or pyrogallol red bleaching). CONCLUSION: The results of this research indicate that there is potential to add value to pawpaw as a functional food source of natural antioxidants, particularly in meat products. © 2017 Society of Chemical Industry.

Phytochemical analysis of ten varieties of pawpaw (Asimina triloba [L.] Dunal) fruit pulp.

Pawpaw (Asimina triloba [L.] Dunal) is a tree fruit with the potential to become a high-value fruit crop, however, its rapid perishability is a significant obstacle. The objective was to determine the phytochemical content and quality characteristics of pawpaw pulp from ten varieties. This study reports for the first time the mass spectral characterization of phenolic acids and flavonoids of pawpaw, which indicated that the predominant polyphenolic compounds were three phenolic acids, protocatechuic acid hexoside, p-coumaroyl hexoside, and 5-O-p-coumaroylquinic acid, and flavonols, particularly (-)-epicatechin, B-type procyanidin dimers and trimers. The relationship between the polyphenolics identified in the current study and future work on polyphenolic oxidase activity will help the process of assessing whether pawpaws should be selected based on potential health benefits, i.e. high polyphenolic content, or increased shelf life in the form of decreased browning that may be afforded pawpaws containing low polyphenolic levels via decreased action of polyphenol oxidase.

Effect of N-acetyl-cysteine on liposomal and muscle model oxidation induced by reactive oxygen, nitrogen, and sulfur.

N-acetyl-cysteine (NAC), a naturally occurring thiol, is found in some fruits and vegetables, sometimes in concentrations higher than glutathione. The objective of this research was to determine the antioxidative effect of NAC in liposomal and muscle models challenged by different oxidizing systems, three that produce reactive oxygen species, two that produce reactive nitrogen species, and two that produce reactive sulfur. The antioxidative effect of cysteine and NAC was compared in the liposomes and NAC and BHT were compared in the muscle homogenates. Lipid hydroperoxides (LOOH), TBARS, and sulfydryls (protein and non-protein) were analyzed. Results indicated that NAC is a more effective inhibitor of lipid oxidation in systems induced by free radicals and reactive nitrogen than those that are induced by peroxides. NAC appears to be at least mildly antioxidative in both liposomal and muscle models, although it did not completely inhibit oxidation in liposomes and generally was not as effective as BHT in the muscle models.

Reactive sulfur species act as prooxidants in liposomal and skeletal muscle model systems.

Reactive sulfur species (RSS) are redox-active sulfur compounds formed under conditions of oxidative stress that may be capable of initiating oxidation reactions. The objective of this research was to determine if two RSS, sulfite radicals and disulfide S-oxides (DSSO), induce oxidation in n-3 liposomes and muscle homogenates. Sulfite radicals and DSSO caused an increase in secondary oxidation products compared to the unoxidized control within 5 min of addition to liposomes. In muscle homogenates, DSSO caused immediate oxidation of sulfhydryls, whereas sulfite radicals caused a slow and linear oxidation of sulfhydryls and lipid. These results suggest that if conditions exist in muscle foods that promote the formation of RSS, they could be a factor in the course of lipid oxidation by either weakening antioxidant defenses or directly oxidizing lipid molecules.

Relating instrumental texture, determined by variable-blade and Allo-Kramer shear attachments, to sensory analysis of rainbow trout, Oncorhynchus mykiss, fillets.

UNLABELLED: Texture is one of the most important quality attributes of fish fillets, and accurate assessment of variation in this attribute, as affected by storage and handling, is critical in providing consistent quality product. Trout fillets received 4 treatments: 3-d refrigeration (R3), 7-d refrigeration (R7), 3-d refrigeration followed by 30-d frozen storage (R3F30), and 7-d refrigeration followed by 30-d frozen storage (R7F30). Instrumental texture of raw and cooked fillets was determined by 3 approaches: 5-blade Allo-Kramer (AK) and variable-blade (VB) attachment with 12 blades arranged in perpendicular (PER) and parallel (PAR) orientations to muscle fibers. Correlation between instrumental texture and sensory hardness, juiciness, elasticity, fatness, and coarseness was determined. Muscle pH remained constant at 6.54 to 6.64. Raw fillets lost 3.66% of their original weight after 30-d frozen storage. After cooking, weight loss further increased to 15.97%. Moisture content decreased from 69.11 to 65.02%, while fat content remained constant at 10.41%. VBPER detected differences in muscle sample strength (P= 0.0019) and demonstrated effect of shear direction reported as maximum force (g force/g sample). AKPER detected differences in energy of shear (g × mm; P= 0.0001). Fillets that received F30 treatments were less extensible. Cooking increased muscle strength and toughness. Force determined by VBPER was correlated with sensory hardness (r= 0.423, P= 0.0394) and cook loss (r= 0.412, P= 0.0450). VB attachment is accurate, valid, and less destructive in fillet texture analysis. PRACTICAL APPLICATION: A new shearing device was validated with sensory analysis. Settings and parameters obtained could be used to define fillet texture quality associated with muscle fiber orientation.

Grape seed extract inhibits lipid oxidation in muscle from different species during refrigerated and frozen storage and oxidation catalyzed by peroxynitrite and iron/ascorbate in a pyrogallol red model system.

The antioxidant effect of grape seed extract (GSE) was determined by assessing the bleaching of pyrogallol red (PGR) by peroxynitrite or iron/ascorbate, and the formation of lipid hydroperoxides (LOOH) and thiobarbituric acid substances (TBARS) in raw or cooked ground muscle during refrigerated or frozen storage. In PGR models, GSE was more effective than gallic acid in inhibiting oxidation. The formation of LOOH and TBARS was inhibited by GSE (0.1% and 1.0%) compared to untreated controls and samples treated with sodium tripolyphosphate. Diethylenetriaminepentaacetic acid (DTPA), alone or in combination with GSE, had no effect on LOOH or TBARS, which provides clues about the possible mechanism of action of GSE. These results show that GSE at concentrations as low as 0.1% is a very effective inhibitor of primary and secondary oxidation products in various muscle systems and has potential as a natural antioxidant in raw and cooked meat systems.

Degradation of γ- and α-tocopherol and formation of 5-nitro-γ-tocopherol induced by peroxynitrite in liposomes and skeletal muscle.

Peroxynitrite, formed from the reaction between nitric oxide and superoxide, can participate in free radical-mediated reactions with cellular components in muscle to (1) initiate lipid oxidation via the production of lipid hydroperoxides, and (2) produce novel nitrated products. 5-Nitro-γ-tocopherol (NGT) is formed by the electrophilic substitution reaction between peroxynitrite and γ-tocopherol. The objective of this research was to examine the utility of NGT as a lipid-phase, peroxynitrite-specific biomarker in muscle foods. NGT was detected when exogenous peroxynitrite was added to liposomes containing γ-tocopherol and homogenates from chicken dark and turkey light muscle with added γ-tocopherol. Detectable levels of NGT were not observed in either minced turkey light muscle stored at -20 °C or chicken dark muscle stored at 4 °C. These results suggest that NGT is not a suitable biomarker to confirm the presence of endogenously produced peroxynitrite in muscle foods.

Potential of peroxynitrite to alter the color of myoglobin in muscle foods.

Superoxide anion and nitric oxide can react to form the highly oxidizing species peroxynitrite. The objective of this research was to determine if peroxynitrite can promote the discoloration of myoglobin under conditions expected in muscle foods. Reagent peroxynitrite (25-100 microM) caused rapid and extensive formation of metmyoglobin from oxymyoglobin with the majority of metmyoglobin formation occurring during the first 5-10 min of incubation. Carbon dioxide caused a small decrease in the ability of peroxynitrite to oxidize oxymyoglobin, and peroxynitrite-promoted conversion of oxymyoglobin to metmyoglobin increased with decreasing pH (5.5-7.0). Differential scanning calorimetry suggested that peroxynitrite caused minimal changes in myoglobin structure. These results indicate that peroxynitrite can promote the conversion of oxymyoglobin to metmyoglobin under the conditions expected in muscle foods.

Nitric oxide synthase activity in muscle foods.

Nitric oxide is enzymatically produced in animals by nitric oxide synthase (NOS). Nitric oxide reacts with superoxide to form peroxynitrite, which initiates oxidative reactions. To assess the potential for nitric oxide formation in muscle, NOS activity was determined in fresh muscle (<8 h post-mortem) from several species under conditions expected in muscle foods. Fresh muscle from all species exhibited NOS activity. NOS activity in muscle was reduced during refrigerated storage. Chicken thigh muscle NOS activity was not affected by pH over the range of 4.5-7.4, but was inhibited by 1 and 2% NaCl. Chicken thigh muscle NOS activity was stimulated at internal cooking temperatures up to 55 °C but was completely inhibited at higher temperatures. Results of this study indicate that post-mortem NOS is only active for the first several days of post-mortem storage.