Concerns have been raised about the safety and tolerability of phytosterol esters due to their vulnerability to oxidation. Herein, oxidation of the unsaturated fatty acid-phytosterol ester, namely β-sitosteryl oleate, was observed in comparison to native β-sitosterol after accelerated storage at 65 °C for 35 days in a bulk oil model system. Depending on the sterol structure, various chemical indices of lipid oxidation, including hydroperoxide value (HPV), thiobarbituric acid reactive substances (TBARS), p-anisidine value (AnV), and 7-keto derivatives, changed at varying rates in both samples. Such indicators for β-sitosteryl oleate appeared to be obtained at higher concentrations than those for β-sitosterol. The first order kinetic was used to describe the losses of β-sitosteryl oleate and β-sitosterol in bulk oil. It was discovered that the β-sitosteryl oleate (k = 0.0202 day-1) underwent oxidative alteration more rapidly than β-sitosterol (k = 0.0099 day-1). Results indicated that physical structure was the principal factor in the determination of storage stability of phytosterol and its ester. Research on antioxidants and storage techniques can be expanded in order to reduce the oxidative loss of phytosterol esters during storage and improve the safety and tolerability of phytosterol esters.
This work comparatively investigated the effects of different levels (0, 1, 3, and 5%, w/w) of cricket protein powder (CP) and soy protein isolate (SPI) on the gel properties of mackerel surimi. Both SPI and CP enhanced the rheological properties of surimi pastes during heating, as indicated by the increase in G' and G″ and the decrease in tan δ. With increasing SPI content, the proteolytic inhibition, gel properties, water-holding capacity, and textural profiles of surimi gel were markedly enhanced. Molecular driving-force results showed that SPI markedly promoted the hydrophobic interaction, while disulfide bonds were dominant in CP-added gel. However, the whiteness of surimi gels tended to decrease with the increased levels of both additives, in particular CP. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that SPI hindered the polymerization of myosin heavy chain while CP participated in the formation of non-disulfide covalent bonds with actin. Fourier transform infrared (FTIR) spectra indicated that CP and SPI did not influence the secondary structure of proteins in surimi. Scanning electron microscopy (SEM) demonstrated that CP or SPI induced the myofibrillar protein to form smoother and compact gel network structures. Overall acceptability of the mackerel surimi gel can be improved by the incorporation of 5% SPI while CP had a negative impact on several parameters. However, CP showed the remarkable ability to prevent the lipid oxidation of the gel after storage at 4 °C for 7 days. Overall, both SPI and CP demonstrated positive impacts on the gelling characteristics of mackerel surimi; however, SPI was more advantageous than CP in terms of the gel-strengthening effect and sensory qualities. This study offered a potential use for plant and insect proteins as functional and nutritional ingredients for the production of dark-fleshed fish surimi.
A thorough investigation of the viability of rice starch conjugation with three different phenolic compounds-gallic acid, sinapic acid, and crude Mon-pu (Glochidion wallichianum Muell Arg) (MP) extract-was conducted using a variety of developed methods which modified the techno-functionality and digestibility of the end product. With and without the aid of ultrasonication (US), phenolic compounds were complexed with hydrothermally pre-gelatinized rice starch prepared using distilled water or plasma-activated water (PAW). The in vitro digestibility, structural features, rheological and thermal properties, and in vitro antioxidant activity of starch-phenolic complexes were evaluated. The US-assisted starch-MP complex in water had the highest complexing index (CI) value (77.11%) and resistant starch (RS) content (88.35%), resulting in a more compact and stable ordered structure. In all complexes, XRD revealed a new minor crystalline region of V-type, which was stabilized by hydrogen bonding as defined by FTIR and H1-NMR. Polyphenols caused a looser gel structure of starch, as imaged by a scanning electron microscope (SEM). Starch-phenolic complexes outperformed other complexes in terms of in vitro antioxidant activity. Gallic acid addition to starch molecules boosted DPPH scavenging activity, notably when synthesized in PAW regardless of US assistance, although having lower CI and RS values than the MP complex. Therefore, this research lays the groundwork for the efficient production of functional food ingredients based on rice starch and polyphenols.
Although dark muscle is currently the most important obstacle in marketing high-quality Indian mackerel (Rastrelliger kanagurta) surimi, reducing washing remains a challenge for long-term surimi production from this species. Herein, the impact of washing cycles (one (W1), two (W2), and three (W3) cycles) with a 1:3 mince to water ratio on the bio-physico-chemical properties, rheology, and gelling ability of mackerel surimi was evaluated. The yield, Ca2+-ATPase activity, TCA-soluble peptide, and myoglobin contents of surimi decreased as the number of washing cycles increased, while lipid removal, reactive SH content, and surface hydrophobicity of surimi increased. Surimi generated by W2 and W3 provided the same rheological patterns and Fourier-transform infrared spectroscopy (FTIR) spectra as unwashed mince, with the highest gel strength and whiteness, as well as the lowest expressible drip, thiobarbituric acid reactive substances (TBARS), and fishy odor. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated the presence of polymerized proteins stabilized by disulfide and other interactions. Using a scanning electron microscope, several concentrated dense areas and distributed pores generated by myofibrillar proteins gel networks were found. Surimi from W2 and W3 appeared to be of similar overall quality, however W2 had a larger yield. As a result of the evaluation of bio-physico-chemical, rheological, and gel-forming capabilities, as well as product yield, W2 may be the best option for producing high-quality surimi from Indian mackerel in a sustainable manner.
The production of bioactive peptides from animal-based raw materials highly depends on enzymatic hydrolysis. Porcine placenta is an underutilized biomass in Thailand's pig farms, yet it is still a source of proteins and beneficial compounds. Porcine placenta could be used as a protein substrate for the production of enzymatic hydrolysate, which could be employed as a functional food ingredient in the future. The goal of this study was to enzymatically produce porcine placenta hydrolysates (PPH) using three commercial enzymes (Alcalase, Flavouzyme, and papain) and evaluate their in vitro antioxidant and antibacterial activity. The degree of hydrolysis (DH) increased as the enzyme load and hydrolysis time increased, but the DH was governed by the enzyme class. The maximum DH was found after using 10% enzyme for 20 min of hydrolysis (36.60%, 31.40%, and 29.81% for Alcalase, Flavouzyme, and papain). Depending on the enzyme type and DH, peptides of various sizes (0.40-323.56 kDa) were detected in all PPH. PPH created with Alcalase had an excellent reducing capacity and metal chelating ability (p < 0.05), whereas PPH made with Flavourzyme and Papain had higher DPPH• and ABTS•+ inhibitory activities (p < 0.05). Papain-derived PPH also had a strong antibacterial effect against Staphylococcus aureus and Escherichia coli, with clear zone values of 17.20 mm and 14.00 mm, respectively (p < 0.05). When PPH was transported via a gastrointestinal tract model system, its antioxidative characteristics were altered. PPH's properties and bioactivities were thus influenced by the enzyme type, enzyme concentration, and hydrolysis time used. Therefore, PPH produced from porcine placenta can be categorized as an antioxidant and antibacterial alternative.
Immobilization can be used to improve the stability of lipases and enhances lipase recovery and reusability, which increases its commercial value and industrial applications. Nevertheless, immobilization frequently causes conformational changes of the lipases, which decrease lipase catalytic activity. in the present work, we synthesized UIO-66 and grafted UIO-66 crystals with proline for immobilization of Candida rugosa lipase (CRL). As indicated by steady-state fluorescence microscopy, grafting of proline onto UIO-66 crystals induced beneficial conformational change in CRL. CRL immobilized on UIO-66/Pro (CRL@UIO-66/Pro) demonstrated higher enzyme activity and better recyclability than that immobilized on UIO-66 (CRL@UIO-66) in both hydrolysis (CRL@UIO-66/Pro: 0.34 U; CRL@UIO-66: 0.15 U) and transesterification (CRL@UIO-66/Pro: 0.93 U; CRL@UIO-66: 0.25 U) reactions. The higher values of kcat and kcat/Km of CRL@UIO-66/Pro also showed that it had better catalytic efficiency as compared to CRL@UIO-66. It is also worth noting that CRL@UIO-66/Pro (0.93 U) demonstrated a much higher transesterification activity as compared to free CRL (0.11 U), indicating that UIO-66/Pro has increased the solvent stability of CRL. Both CRL@UIO-66 and CRL@UIO-66/Pro were also used for the fabrication of biosensors for nitrofen with a wide linear range (0-100 μM), lower limit of detection, and good recovery rate.
Milk processing technology disrupts milk fat globule membrane (MFGM) structures and decreases the phospholipid content in MFGM. The present study aimed to evaluate the effects of homogenization, thermal treatment, and cold storage on milk phospholipids. A total of 175 phospholipid molecular species were identified and quantified. Phosphatidylcholine was the most abundant phospholipid, and sphingomyelin accounted for only a small amount of phospholipid in bovine milk. In addition, a total of 37 plasmalogens (167.5 μg/mL) were identified in bovine milk with lysophosphatidylcholine plasmalogen being the most abundant. Processing technologies decreased the phospholipid content with both boiled and frozen milk demonstrating the highest reduction. Compared to raw milk, only 70% of phospholipid remained in frozen milk. Both S-plot and volcano-plot showed that heat treatment and subsequent cold storage decreased the phosphatidylserine and lysophospholipid contents.
Indigenous southern Thai non-glutinous rice varieties Kaab Dum, Khai Mod Rin, Yar Ko, Yoom Noon, and Look Lai made under four different processing conditions, white rice, brown rice, germinated brown rice, and rice grass, were assessed for antioxidant components and in vitro antioxidative activities. According to the findings, rice’s antioxidant components and antioxidant activity were considerably impacted by both variety and processing. High levels of total extractable phenolic compounds (164−314 mg gallic acid equivalent (GAE)/kg, dry weight (dw)) and carotenoid (0.92−8.65 mg/100 g, dw) were found in all rice varieties, especially in rice grass and germinated brown rice, indicating that milling to generate white rice had an adverse effect on those components. Additionally, after germination, a higher γ-oryzanol concentration (9−14 mg/100 g, dw) was found. All rice varieties had higher ascorbic acid, phenolic compound, and carotenoid contents after sprouting. Overall, Yoom Noon rice grass had the highest total extractable phenolic content (p < 0.05). The rice grass from Yoom Noon/Look Lai/Kaab Dum had the highest ascorbic acid content (p < 0.05). The total carotenoid concentration of Look Lai rice grass was the highest, and Yoom Noon’s germinated brown rice had the highest γ-oryzanol content (p < 0.05). All rice varieties’ aqueous extracts had remarkable ABTS free radical scavenging activity, with Khai Mod Rin reaching the highest maximum value of 42.56 mmol Trolox equivalent/kg dw. Other antioxidant mechanisms, however, were quite low. Compared to germinated brown rice, brown rice, and white rice, rice grass often tended to have stronger antioxidant activity. Yar Ko rice grass was found to have the highest DPPH free radical scavenging activity (3.8 mmol Trolox equivalent/kg dw) and ferric reducing antioxidant power (FRAP) (4.6 mmol Trolox equivalent/kg dw) (p < 0.05). Khai Mod Rice grass had the most pronounced metal chelation activity (1.14 mmol EDTA equivalent/kg dw) (p < 0.05). The rice variety and processing conditions, therefore, influenced the antioxidant compounds and antioxidative properties of Thai indigenous rice. The results can be used as a guide to select the optimal rice variety and primary processing in order to satisfy the needs of farmers who want to produce rice as a functional ingredient and to promote the consumption of indigenous rice by health-conscious consumers.
The stimulus-responsive regulation of enzyme catalytic activity and selectivity provides a new opportunity to extend the functionality and efficiency of immobilized enzymes. This work aims to design and synthesize a thermo-switchable enzyme@MOF for size-selective biocatalysis and biosensing through the immobilization of Candida rugosa lipase (CRL) within ZIF-8 functionalized with thermally responsive polymer, poly(N-isopropylacrylamide) (PNIPAM) (CRL@ZIF-8-PNIPAM). Unlike free CRL, which does not demonstrate substrate selectivity, we can reversibly tune the pore size of the ZIF-8-PNIPAM nanostructures (open pores or blocked pores) through temperature stimulus and subsequently modulate the substrate selectivity of CRL@ZIF-8-PNIPAM. CRL@ZIF-8-PNIPAM had the highest hydrolytic activity for small molecules (12 mM p-nitrophenol/mg protein/min, 4-nitrophenyl butyrate (p-NP Be)) and the lowest hydrolytic activity for large molecules (0.16 mM p-nitrophenol/mg protein/min, 4-nitrophenyl palmitate (p-NP P)). In addition, CRL@ZIF-8-PNIPAM demonstrated thermo-switchable behavior for large molecules (p-NP P). The p-NP P hydrolytic activity of CRL@ZIF-8-PNIPAM was significantly lower at 40 °C (blocked pores) than at 27 °C (open pores). However, the transition of blocked pores and open pores is a gradual process that resulted in a delay in the "thermo-switchable" catalytic behavior of CRL@ZIF-8-PNIPAM during thermal cycling. CRL@ZIF-8-PNIPAM was also successfully used for the fabrication of electrochemical biosensors for the selective biosensing of pesticides with different molecular sizes.
The present study aims to design a milk fat globule membrane (MFGM)-inspired structured membrane (phospholipid- and protein-rich) for microencapsulation of docosahexaenoic acid (DHA) oil. DHA-enriched oil emulsions were prepared using different ratios of sunflower phospholipid (SPL), proteins [whey protein concentrate (WPC), soy protein isolate (SPI), and sodium caseinate (SC)], and maltodextrin and spray-dried to obtain DHA microcapsules. The prepared DHA oil emulsions have nanosized particles. SPLs were found to affect the secondary structure of WPC, which resulted in increased exposure of the protein hydrophobic site and emulsion stability. SPL also reduced the surface tension and viscosity of the DHA oil emulsions. In vitro digestion of the spray-dried DHA microcapsules showed that they were able to effectively resist gastric proteolysis and protect their bioactivity en route to the intestine. The DHA microcapsules have a high lipid digestibility in the small intestine with a high DHA hydrolysis efficiency (74.3%), which is higher than that of commercial DHA microcapsules.
Gestational diabetes mellitus (GDM) is a prevalent metabolic disorder during pregnancy that alters the metabolites in human milk. Integrated Gas Chromatography-Mass Spectrometry (GC-MS) and Liquid Chromatography-Mass Spectrometry (LC-MS) were employed for comprehensive identification and comparison of metabolites in mature human milk (MHM) from women with and without GDM. A total of 268 differentially expressed metabolites (DEMs) were identified. Among these, linoleic acid, arachidonic acid, 9R-HODE and L-glutamic acid were significantly elevated and 12,13-DHOME was significantly decreased in MHM of women with GDM. These metabolites are significantly enriched in linoleic acid metabolism, fatty acid biosynthesis, galactose metabolism and ABC transporters pathways. Disorders in these metabolic pathways are associated with insulin resistance and poor glucose metabolism indicating these conditions may persist postpartum.