Exploration of the pearl millet phospholipase gene family to identify potential candidates for grain quality traits.

BACKGROUND: Phospholipases constitute a diverse category of enzymes responsible for the breakdown of phospholipids. Their involvement in signal transduction with a pivotal role in plant development and stress responses is well documented. RESULTS: In the present investigation, a thorough genome-wide analysis revealed that the pearl millet genome contains at least 44 phospholipase genes distributed across its 7 chromosomes, with chromosome one harbouring the highest number of these genes. The synteny analysis suggested a close genetic relationship of pearl millet phospholipases with that of foxtail millet and sorghum. All identified genes were examined to unravel their gene structures, protein attributes, cis-regulatory elements, and expression patterns in two pearl millet genotypes contrasting for rancidity. All the phospholipases have a high alpha-helix content and distorted regions within the predicted secondary structures. Moreover, many of these enzymes possess binding sites for both metal and non-metal ligands. Additionally, the putative promoter regions associated with these genes exhibit multiple copies of cis-elements specifically responsive to biotic and abiotic stress factors and signaling molecules. The transcriptional profiling of 44 phospholipase genes in two genotypes contrasting for rancidity across six key tissues during pearl millet growth revealed a predominant expression in grains, followed by seed coat and endosperm. Specifically, the genes PgPLD-alpha1-1, PgPLD-alpha1-5, PgPLD-delta1-7a, PgPLA1-II-1a, and PgPLD-delta1-2a exhibited notable expression in grains of both the genotypes while showing negligible expression in the other five tissues. The sequence alignment of putative promoters revealed several variations including SNPs and InDels. These variations resulted in modifications to the corresponding cis-acting elements, forming distinct transcription factor binding sites suggesting the transcriptional-level regulation for these five genes in pearl millet. CONCLUSIONS: The current study utilized a genome-wide computational analysis to characterize the phospholipase gene family in pearl millet. A comprehensive expression profile of 44 phospholipases led to the identification of five grain-specific candidates. This underscores a potential role for at least these five genes in grain quality traits including the regulation of rancidity in pearl millet. Therefore, this study marks the first exploration highlighting the possible impact of phospholipases towards enhancing agronomic traits in pearl millet.

Exploring selection signatures in the divergence and evolution of lipid droplet (LD) associated genes in major oilseed crops.

BACKGROUND: Oil bodies or lipid droplets (LDs) in the cytosol are the subcellular storage compartments of seeds and the sites of lipid metabolism providing energy to the germinating seeds. Major LD-associated proteins are lipoxygenases, phospholipaseD, oleosins, TAG-lipases, steroleosins, caleosins and SEIPINs; involved in facilitating germination and enhancing peroxidation resulting in off-flavours. However, how natural selection is balancing contradictory processes in lipid-rich seeds remains evasive. The present study was aimed at the prediction of selection signatures among orthologous clades in major oilseeds and the correlation of selection effect with gene expression. RESULTS: The LD-associated genes from the major oil-bearing crops were analyzed to predict natural selection signatures in phylogenetically close-knit ortholog clusters to understand adaptive evolution. Positive selection was the major force driving the evolution and diversification of orthologs in a lineage-specific manner. Significant positive selection effects were found in 94 genes particularly in oleosin and TAG-lipases, purifying with excess of non-synonymous substitution in 44 genes while 35 genes were neutral to selection effects. No significant selection impact was noticed in Brassicaceae as against LOX genes of oil palm. A heavy load of deleterious mutations affecting selection signatures was detected in T-lineage oleosins and LOX genes of Arachis hypogaea. The T-lineage oleosin genes were involved in mainly anther, tapetum and anther wall morphogenesis. In Ricinus communis and Sesamum indicum > 85% of PLD genes were under selection whereas selection pressures were low in Brassica juncea and Helianthus annuus. Steroleosin, caleosin and SEIPINs with large roles in lipid droplet organization expressed mostly in seeds and were under considerable positive selection pressures. Expression divergence was evident among paralogs and homeologs with one gene attaining functional superiority compared to the other. The LOX gene Glyma.13g347500 associated with off-flavor was not expressed during germination, rather its paralog Glyma.13g347600 showed expression in Glycine max. PLD-α genes were expressed on all the tissues except the seed,δ genes in seed and meristem while ß and γ genes expressed in the leaf. CONCLUSIONS: The genes involved in seed germination and lipid metabolism were under strong positive selection, although species differences were discernable. The present study identifies suitable candidate genes enhancing seed oil content and germination wherein directional selection can become more fruitful.

Oxidative Stability of Side-Streams from Cod Filleting-Effect of Antioxidant Dipping and Low-Temperature Storage.

Currently, side-streams (e.g., head, backbone, tail, and intestines) generated in the fish processing industry often end up as low-value products for feed applications or even as waste. In order to upcycle such side-streams, they need to be preserved to avoid oxidative degradation of the lipids between the generation point and the valorization plant. In the cod filleting industry, three main solid side-streams: viscera, heads, and backbones, are obtained. Hence, this study aimed to identify the most efficient antioxidant for preserving the cod side-streams using a dipping-based strategy prior to pre-valorization storage at low temperatures (ice and frozen storage). The dipping solutions evaluated contained: (i) a lipophilic rosemary extract (0.05% and 0.2% in 0.9% NaCl), (ii) Duralox MANC (a mixture of rosemary extract, ascorbic acid, tocopherols, and citric acid; 2% in 0.9% NaCl), and (iii) NaCl (0.9%) w/w solution. One group was not dipped. No dipping and dipping in NaCl were included as controls. The results showed a positive effect of dipping with solutions containing antioxidants as measured by peroxide value (PV), TBA-reactive substances (TBARS), and sensory profiling, e.g., rancid odor. Moreover, the oxidative stability increased with decreased storage temperature. The cod side-streams were in general most efficiently preserved by Duralox MANC, followed by the lipophilic rosemary extract (0.2%), compared to no dipping and dipping in NaCl solution and the lower concentration of the lipophilic rosemary extract (0.05%). The efficiency of the antioxidant treatments was independent of the side-stream fraction and storage temperature. Thus, using antioxidant dipping combined with low temperature storage is an efficient preservation method for maintaining the quality of the lipids in cod solid side-streams during their pre-valorization storage.

Genetic and genomic approaches to address rapid rancidity of rice bran.

Rice bran is an invaluable by-product of paddy processing industry. It is rich in minerals, protein, lipids, and crude fiber. In addition, it also possesses compounds with anti-oxidant, anti-allergic, anti-diabetic, and anti-cancer properties. It forms a basis for the extraction of rice bran oil and preparation of various functional foods with health benefits and potential to prevent chronic health issues. Nevertheless, the rapid deterioration of bran upon storage acts as a major limitation in exploiting the full potential of rice bran. In this review, we have discussed three strategies to address rapid rancidity of rice bran and enhance its shelf life and storability vis-a-vis emphasizing the importance of rice bran in terms of its nutritional composition. One strategy is through exploitation of the null mutations in the genes governing lipases and lipoxygenases leading to nonfunctional enzymes (enzyme deficient approach), another strategy is through reducing the PUFA content that is more prone to oxidation (substrate deficient approach) and a third strategy is through enhancing the antioxidant content that effectively terminate the lipid peroxidation by donating the hydrogen atom.

Optimisation of Healthy-Lipid Content and Oxidative Stability during Oil Extraction from Squid (Illex argentinus) Viscera by Green Processing.

Green extraction was applied to Argentinean shortfin squid (Illex argentinus) viscera, consisting of a wet pressing method including a drying step, mechanic pressing, centrifugation of the resulting slurry, and oil collection. To maximise the oil yield and ω3 fatty acid content and to minimise the oil damage degree, a response surface methodology (RSM) design was developed focused on the drying temperature (45-85 °C) and time (30-90 min). In general, an increase of the drying time and temperature provided an increase in the lipid yield recovery from the viscera. The strongest drying conditions showed a higher recovery than 50% when compared with the traditional chemical method. The docosahexaenoic and eicosapentaenoic acid contents in the extracted oil revealed scarce dependence on drying conditions, showing valuable ranges (149.2-166.5 and 88.7-102.4 g·kg-1 oil, respectively). Furthermore, the values of free fatty acids, peroxides, conjugated dienes, and ω3/ω6 ratio did not show extensive differences by comparing oils obtained from the different drying conditions. Contrary, a polyene index (PI) decrease was detected with increasing drying time and temperature. The RSM analysis indicated that optimised drying time (41.3 min) and temperature (85 °C) conditions would lead to 74.73 g·kg-1 (oil yield), 1.87 (PI), and 6.72 (peroxide value) scores, with a 0.67 desirability value.

Effectiveness of the Natural Antioxidant 2,4,4'-Trihydroxychalcone on the Oxidation of Sunflower Oil during Storage.

This study aimed to investigate the effectiveness of 2,4,4'-trihydroxychalcone as a natural antioxidant on the oxidation of sunflower oil during an 88-day storage period and to compare its strength with the synthetic antioxidant butylated hydroxytoluene (BHT). Seven groups of the sunflower oil samples were prepared: pure oil (control), oil treated with different concentrations (100, 500, and 1000 ppm) of 2,4,4'-trihydroxychalcone, and oil treated with different concentrations (100, 500, and 1000 ppm) of BHT. Specific parameters, namely, the peroxide value (PV), acid value (AV), p-anisidine value (p-AnV), thiobarbituric acid reactive substance (TBARS) value and total oxidation (TOTOX) value were used to assess the extent of the deterioration of the oil by estimating the primary and secondary oxidation products. The results showed that 2,4,4'-trihydroxychalcone effectively decreased the production of the primary and secondary oxidation products of sunflower oil during storage, as indicated by reductions in the PVs, AVs, p-AnVs, TBARS values and TOTOX values of the sunflower oil. When compared to BHT, 2,4,4'-trihydroxychalcone showed either a similar or stronger effect in inhibiting the primary and secondary oxidation products. These findings suggest that, 2,4,4'-trihydroxychalcone is a suitable natural alternative to synthetic antioxidants to improve the oxidative stability of sunflower oil.

Exploring genetic dependence of lipase activity to improve the quality of whole-grain wheat.

BACKGROUND: Whole-grain wheat flour is facing the quality challenge of lipid rancidity, which decreases its nutritional, sensory, and technological properties. One of the major causes of lipid rancidity is endogenous esterases and lipases. This study evaluated 66 European wheat varieties grown at a single site over three years (2014, 2015, and 2016). RESULTS: The 66 wheat varieties showed up to threefold variance on esterase and lipase activities. Wheat varieties that are suitable for lipid-stable whole-grain products ('Julius', 'Lona', and 'Banquet') were selected according to their consistently low esterase and lipase activities. The 3-year mean-based broad-sense heritability of esterase and lipase was 0.75 and 0.44 respectively. CONCLUSIONS: The findings indicate great genetic dependence of both esterase and lipase activities in wheat. The moderate to high heritability brings a new prospect of breeding selection of low-lipase-activity wheat for stable whole-grain products. This result will improve the use of wheat as raw material, benefit cultivation selection, and provide consumers with better quality products. © 2020 Society of Chemical Industry.

Fresh and stored copra meal in meat quail diets.

The aim of the current study was to evaluate the addition of fresh and stored copra meal to the diet of meat quails. Two hundred eighty-seven-day-old male and female quails were distributed in a completely randomized design, with five treatments, in a 2 × 2 + 1factorial arrangement. Two copra meal types (fresh and stored) at inclusion levels of 12.5% and 25%, respectively, were added to a corn-soybean meal-based diet, with seven replicates, of eight quails each. Copra meal acidity index recorded oleic acid percentage increase from 0.47 to 3.03% after six storage months. However, regardless of storage type, its addition to quails' diet resulted in higher values of metabolizable energy, in lower feed intake and better feed conversion than corn-soybean meal diet. Copra meal addition to quails' diet did not affect carcass traits, liver and pancreas relative weight, and bone growth and quality. Although copra meal storage for 180 days resulted in higher free fatty acid content in the provided feed, it can be used fresh or after storage, in diet of meat quails from 7 to 42 days of age up to 25%.

Mechanisms involved in hemoglobin-mediated oxidation of lipids in washed fish muscle and inhibitory effects of phospholipase A2.

BACKGROUND: Hemoglobin (Hb) is a lipid oxidation promoter in fish muscle. Phospholipase A2 (PLA2; EC 3.1.1.4) is linked to an increased resistance to lipid oxidation of frozen-thawed cod fillets via an unknown mechanism. The present study aimed to investigate the mechanism of Hb-mediated lipid oxidation with a focus on ferryl Hb and methemoglobin (metHb), the pro-oxidative Hb species, and to examine how porcine pancreatic PLA2 inhibits Hb-mediated lipid oxidation in washed cod muscle (WCM). Lipid hydroperoxides (LOOHs) and thiobarbituric acid reactive substances (TBARS) were measured as primary and secondary lipid oxidation products, respectively. The formation of metHb and ferryl Hb was also monitored. RESULTS: Ferryl Hb and metHb formed during the Hb-mediated lipid oxidation. PLA2 inhibited the formation of LOOHs and TBARS and suppressed the formation of metHb and ferryl Hb. WCM was pre-oxidized by hemin to increase the amount of LOOHs. PLA2 promoted the depletion of LOOHs in the pre-oxidized WCM with limited TBARS formation at the expense of the heme moiety of Hb. CONCLUSION: The results of the present study suggest that ferryl Hb may play a role in Hb-mediated lipid oxidation and that PLA2 from pig pancreas may work together with Hb as a novel antioxidant with an ability to remove pre-formed LOOHs from a lipid substrate. © 2017 Society of Chemical Industry.

Oxidative deterioration of pork during superchilling storage.

BACKGROUND: In superchilling (SC), meat is kept at temperatures around 1 °C below its initial freezing point, leading to a significant increase in shelf life. This study aimed to address the oxidative changes taking place in pork loins during prolonged storage at SC temperature. Loins were stored either at chilling (CH) conditions (2-4 °C) for 4 weeks or at SC temperature (around -1 °C) for 12 weeks. RESULTS: Storage at SC temperature diminished the rate of lipid and protein oxidation and discoloration in pork loins, so that final levels of most oxidation products and instrumental color values after 12 weeks of SC storage were similar to those after 4 weeks at CH conditions. However, hexanal content peaked by the end of SC storage, pointing to a potential accumulation of compounds from lipid oxidation during SC storage. CONCLUSION: SC storage of pork slows down the rate of lipid and protein oxidation. However, accumulation of volatile compounds from lipid oxidation could be a limiting factor for shelf life. © 2017 Society of Chemical Industry.

Interaction between rancidity and organoleptic parameters of anchovy marinade (Engraulis encrasicolus L. 1758) include essential oils.

This study was carried out to evaluate the lipid oxidation and sensory attributes of anchovy marinated with 10% NaCl+4% alcohol vinegar+0.2% citric acid solution and 0.1% different essential oils. Group A Control: only sunflower seed oil, Group B: sunflower seed oil+0.1% rosemary oil, Group C: sunflower seed oil+0.1% coriander oil, Group D: sunflower seed oil+0.1% laurel oil and Group E: sunflower seed oil+0.1% garlic oil. During storage, lipid oxidation as indicated by the 2-thiobarbituric acid reactive substances (TBARs) values of the control group were significantly higher than the other groups containing essential oils. The results showed that the essential oils have retarding effect on lipids oxidation. This effect was the highest in laurel oil during initial 3 months; and it was similar to laurel oil and rosemary oil in the fourth month; in all the essential oil added groups in 6 month. L*(brightness) values were similar for all groups in first fourth months but, at the last 2 months, group using laurel oil was found better. Yellowness (b*) was similar in all groups during the intial 3 months whereas, after that lower values in the groups that used laurel and rosemary oils were detected. The study concluded that marination with 0.1% laurel oil of anchovy can retard lipid oxidation and improve the sensory attributes of the product during refrigerated storage.

Inhibition of Shewanella spp. growth by Syzygium australe and Syzygium luehmannii extracts: natural methods for the prevention of fish spoilage.

Syzygium australe and Syzygium luehmannii fruit and leaf were investigated for their ability to inhibit Shewanella spp. growth. Extracts of both Syzygium spp. displayed potent growth inhibitory properties against all Shewanella spp. tested in disc diffusion and liquid diffusion assays. In general, S. australe extracts were more potent inhibitors of Shewanella spp. growth, and the fruit extracts were generally better than the corresponding leaf extracts. The methanolic S. australe fruit extract was a particularly potent inhibitor of all Shewanella spp. growth, with MIC values as low as 87 µg/mL. The aqueous and ethyl acetate S. australe fruit extracts were similarly potent inhibitors of Shewanella spp. growth, albeit with slightly higher MIC values. Several other Syzygium spp. extracts also were potent bacterial growth inhibitors, albeit with MIC values generally >1000 µg/mL. The most potent S. australe fruit extracts were nontoxic in the Artemia franciscana bioassay, with LC50 values substantially >1000 µg/mL. The potent bacterial growth inhibitory activity and lack of toxicity of the S. australe fruit extracts indicate their potential as natural fish and seafood preservatives.

Rapid and non-destructive determination of rancidity levels in butter cookies by multi-spectral imaging.

BACKGROUND: Rancidity is an important attribute for quality assessment of butter cookies, while traditional methods for rancidity measurement are usually laborious, destructive and prone to operational error. In the present paper, the potential of applying multi-spectral imaging (MSI) technology with 19 wavelengths in the range of 405-970 nm to evaluate the rancidity in butter cookies was investigated. RESULTS: Moisture content, acid value and peroxide value were determined by traditional methods and then related with the spectral information by partial least squares regression (PLSR) and back-propagation artificial neural network (BP-ANN). The optimal models for predicting moisture content, acid value and peroxide value were obtained by PLSR. The correlation coefficient (r) obtained by PLSR models revealed that MSI had a perfect ability to predict moisture content (r = 0.909), acid value (r = 0.944) and peroxide value (r = 0.971). CONCLUSION: The study demonstrated that the rancidity level of butter cookies can be continuously monitored and evaluated in real-time by the multi-spectral imaging, which is of great significance for developing online food safety monitoring solutions.

Exploration of markers in oxidized rancidity walnut kernels based on lipidomics and volatolomics.

Walnut kernels are prone to oxidation and rancidity due to their rich lipid composition, but the existing evaluation indicators are not sensitive enough to promote their industrial development. This study aims to investigate the potential markers in oxidative rancidity walnut kernels using lipidomics and volatolomics. The results showed that the antioxidant capacity of walnut kernels significantly decreased after oxidation, with the decreasing of total phenolic content from 36276.34 mg GAE/kg to 31281.53 mg GAE/kg, the DPPH and ABTS free radical scavenging activity from 89.25% to 73.54%, and 61.69% to 43.73%, respectively. The activities of lipoxygenase (LOX) and lipase (LPS) increased by 6.08-fold and 0.33-fold, respectively. By combining volatolomics and chemometrics methods, it was found that significant differences existed in the content of hexanal, caproic acid, 1-pentanol, (E)-2-octenal, and 2-heptanenal before and after walnut kernel oxidation (VIP > 1). Based on the results of lipidomics, it can be concluded that the above five compounds can serve as characteristic markers for walnut kernel oxidative rancidity, mainly produced through glycerol phospholipid (GPL), glyceride, linoleic acid (LA), and α-linolenic acid (ALA) metabolism pathways. Possible mechanisms of lipid degradation in oxidized walnut kernels were also proposed, providing technical support for the storage, preservation, and high-value utilization of walnut kernels.

Storage and time course effects on the quality of oil extracted from Phyllanthus amarus Schumach and Annona muricata Linn and their antidiabetic potentials.

With the advent of modern technology, advancements in processing and storage techniques, and increasing medical knowledge, people are becoming aware of deterioration in the quality of medicinal products due to storage methods and time. In most cases, herbal products are not consumed immediately after production; as such, improper storage can result in physical, chemical, and microbiological changes. The study evaluated the effect of storage methods and time on the quality of oil extracted from Phyllanthus amarus Schumach and Annona muricata Linn and assessed their antidiabetic and antioxidative effects. Plants were air-dried, pulverized, and then subjected to Soxhlet extraction in petroleum ether. The oil was evaluated for phytochemical constituents and the effects of time and storage methods on its physicochemical properties. Characterization of the oil was done by spectroscopic techniques. Oils from both plants contained tannins, flavonoids, alkaloids, steroids, glycosides, terpenoids, phlobotannins, resins, reducing sugar, phenols, and saponins in different proportions. The oil from A. muricata had higher phenolic (3.11 ± 0.31 mg GAE/g), flavonoid (11.82 ± 0.08 mg QUE/g), alkaloid (16.37 ± 0.56 mg APE/g), and tannin (7.13 ± 0.47 mg CE/g) contents than the oil from P. amarus, which had 0.54 ± 0.08 mg GAE/g, 7.83 ± 0.13 mg QUE/g, 9.87 ± 0.15 mg APE, and 3.16 ± 0.12 mg CE/g for total phenolic, flavonoids, alkaloids, and tannins, respectively. Initial acid, iodine, peroxide, and saponification values recorded for P. amarus were 5.63 ± 0.82 mg KOH/g, 97.17 ±0.53 Wijis, 9.31 ± 0.15 mEq/kg, and 116.11 ± 0.74 mg KOH/g, respectively, significantly different from those of A. muricata , which had values of 1.17 ± 0.08 mg KOH, 76.23 ± 0.03 Wijis, 6.75 ± 0.47 mEq/kg, and 193.31 ± 0.52 mg KOH/g, respectively. FT-IR characterization of the oils revealed the presence of carboxylic acid, alkyl, alkene, alkane, haloalkane, aldehyde, aromatic amine, α-unsaturated and ß-unsaturated esters, and phenol functional groups. P. amarus oil inhibited α-amylase (IC50 0.17 ± 0.03 mg/ml), α-glucosidase (IC50 0.64 ± 0.03 mg/ml), and xanthine oxidase (0.70 ± 0.01 mg/ml) to a greater extent than A. muricata oil, with IC50 values of 0.43 ± 0.05 mg/ml (α-amylase), 2.25 ± 0.31 mg/ml (α-glucosidase), and 0.78 ± 0.07 mg/ml (xanthine oxidase). This study showed that oils from the tested plants have low rancidity with a moderate shelf life. The extracts contained essential phytoconstituents that significantly inhibited α-glucosidase and xanthine oxidase. These effects of the oil indicate their potential to prevent diabetes, gout, and oxidative stress. Consequently, the supply of P. amarus and A. muricata in homemade diets is strongly encouraged for healthy living.

A digital image-based colorimetric method for measuring free acidity in edible vegetable oils.

The standard method used to quantify free acidity (FA) in vegetable oil is neutralization titration, which requires many toxic chemicals and depends on an analyst's experience in detecting endpoints. Here, a digital image colorimetry (DIC) method using a smartphone camera was developed to measure FA in vegetable oils. A cupric acetate solution was used to produce the colorimetric reaction. The coloured solutions were imaged, and R values (from the RGB colour system) were calibrated against the respective FAs in the standards. The FA values of the samples were determined by standard addition calibration. These results were compared to measurements of FA obtained by the standard titrimetric method. An excellent correlation was obtained, with an R2 of 0.98 and a mean absolute error of 0.06%. The chemicals needed for analysis were reduced by approximately 90%. Thus, DIC is a less subjective and more economical method for determining FA in vegetable oils.

Lipidomics analysis unveils the dynamic alterations of lipid degradation in rice bran during storage.

Recent explorations into rice bran oil (RBO) have highlighted its potential, owing to an advantageous fatty acid profile in the context of health and nutrition. Despite this, the susceptibility of rice bran lipids to oxidative degradation during storage remains a critical concern. This study focuses on the evolution of lipid degradation in RBO during storage, examining the increase in free fatty acids (FFAs), the formation of oxylipids, and the generation of volatile secondary oxidation products. Our findings reveal a substantial rise in FFA levels, from 109.55 to 354.06 mg/g, after 14 days of storage, highlighting significant lipid deterioration. Notably, key oxylipids, including 9,10-EpOME, 12,13(9,10)-DiHOME, and 13-oxoODE, were identified, with a demonstrated positive correlation between total oxylipids and free polyunsaturated fatty acids (PUFAs), specifically linoleic acid (LA) and α-linolenic acid (ALA). Furthermore, the study provides a detailed analysis of primary volatile secondary oxidation products. The insights gained from this study not only sheds light on the underlying mechanisms of lipid rancidity in rice bran but also offers significant implications for extending the shelf life and preserving the nutritional quality of RBO, aligning with the increasing global interest in this high-quality oil.

Effect of using biodegradable film constituting red grape anthocyanins as a novel packaging on the qualitative attributes of emergency food bars during storage.

This study presents a novel packaging film based on whey protein isolate/κ-carrageenan (WC) with red grape pomace anthocyanins (RGA) to investigate its impact on some qualitative attributes of emergency food bars (EFBs) for 6 months at 38°C. Increasing the RGA dose in WC films from 5% (WCA5) to 10% (WCA10) reduced hydrogen bonding between polymers and polymer homogeneity in the matrix according to FTIR and SEM. Tensile strength slightly declined in WCA5 from 7.47 ± 0.26 to 6.97 ± 0.12, while elongation increased from 27.74 ± 1.36 to 32.36 ± 1.25% compared to WC film. The maximum weight loss temperature (TM) increased by incorporating 5 wt% RGA from 182.95°C to 244.36°C, whereas TM declined to 187.19°C in WCA10 film. WVP and OTR slightly changed in WCA5 (from 7.83 ± 0.07 and 2.57 ± 0.18 to 8.41 ± 0.03 g H2O.m/m2.Pa.s × 10-9 and 1.79 ± 0.32 cm3 O2/m2.d.bar, respectively), but significantly impaired in WCA10 compared to WC film. WCA5 and WCA10 films had high AA%, 68.77%, and 79.21%, respectively. WCA10 film presented great antimetrical properties against Staphylococcus aureus with an inhibition zone of 6.00 mm. The light transmission of RGA-contained films in the UV spectrum was below 10%. The WCA5 film effectively restrained moisture loss and hardness increment until the end of the storage period, which were 14.33% and 28.76%, respectively, compared to day 0. Antioxidant films provided acceptable resistance against oxidation to EBF treatment. Sensory panels scored WCA5 and WCA10 higher in overall acceptance with 5.64 and 5.40 values, respectively, while complaining about the hardness of OPP treatment. The results of this investigation demonstrated that incorporating RGA, preferably 5 wt%, into WC-based film effectively improved the qualitative properties of EFB during the 6-month shelf life. This film might be a promising alternative for packaging light and oxygen-sensitive food products.

Identification of cheese rancidity-related lipases in Aspergillus oryzae AHU 7139.

The adjunct product with enzymatic activity from Aspergillus oryzae is beneficial for flavor enrichment in the ripened cheese. However, an excessive lipolytic reaction leads to the release of volatile free fatty acids. Accordingly, a strong off-flavor (i.e., rancidity) has been detected when A. oryzae AHU 7139 is used. To identify the rancidity-related lipase from this strain, we evaluated the substrate specificity and lipase distribution using five mutants cultured on a whey-based solid medium under different initial pH conditions. The results showed a higher diacylglycerol lipase activity than triacylglycerol lipase activity. Moreover, an initial pH of 6.5 for the culture resulted in higher lipolytic activity than a pH of 4.0, and most of the activity was found in the extracellular fraction. Based on the gene expression analysis by real-time polymerase chain reaction and location and substrate specificity, five genes (No. 1, No. 19, mdlB, tglA, and cutL) were selected among 25 annotated lipase genes to identify the respective knockout strains. Because ΔtglA and ΔmdlB showed an outstanding involvement in the release of free fatty acids, these strains were applied to in vitro cheese curd experiments. In conclusion, we posit that triacylglycerol lipase (TglA) plays a key role as the trigger of rancidity and the resulting diglycerides have to be exposed to diacylglycerol lipase (MdlB) to stimulate rancidity in cheese made with A. oryzae AHU 7139. This finding could help screen suitable A.oryzae strains as cheese adjuncts to prevent the generation of the rancid-off flavor.

Lipidomics analysis of rice bran during storage unveils mechanisms behind dynamic changes in functional lipid molecular species.

Rice bran, recognized for its rich lipids and health-beneficial bioactive compounds, holds considerable promise in applications such as rice bran oil production. However, its susceptibility to lipid hydrolysis and oxidation during storage presents a significant challenge. In response, we conducted an in-depth metabolic profiling of rice bran over a storage period of 14 days. We focused on the identification of bioactive compounds and functional lipid species (25 acylglycerols and 53 phospholipids), closely tracking their dynamic changes over time. Our findings revealed significant reductions in these lipid molecular species, highlighting the impact of rancidity processes. Furthermore, we identified 19 characteristic lipid markers and elucidated that phospholipid and glycerolipid metabolism were key metabolic pathways involved. By shedding light on the mechanisms driving lipid degradation in stored rice bran, our study significantly advanced the understanding of lipid stability. These information provided valuable insights for countering rancidity and optimizing rice bran preservation strategies.