Changes In Secondary Structure Research Articles

Interactions with peanut protein isolate regulate the bioaccessibility of cyanidin-3-O-glucoside: Multispectral analysis, simulated digestion, and molecular dynamic simulation

Anthocyanins are susceptible to degradation owing to environmental factors. Combining them with proteins can improve their stability; however, the interaction mechanism is difficult to elucidate. This study used multispectral and molecular dynamics simulations and molecular docking methods to investigate the interaction mechanism between peanut protein isolate (PPI) and cyanidin-3-O-glucoside (C3G). The UV absorption peak and PPI turbidity increased, while the fluorescence intensity decreased with greater C3G content. Protein secondary structure changes suggested that PPI and C3G coexisted in spontaneous covalent and non-covalent interactions via static quenching. The complex structures were stable over time and C3G stably bound to the peanut globulin Ara h 3 cavity through hydrogen bonding and hydrophobic interactions. Furthermore, PPI enhanced the C3G antioxidant activity and bioaccessibility by increasing its retention rate during in-vitro simulated digestion. This study elucidates the binding mechanism of PPI and C3G and provides insight into applications of the complex in food development.

Probing amikacin interaction with albumin: A combined multispectral and molecular docking investigation

The fate of a drug administered to a living organism depends on the pharmacological and pharmacokinetic behavior of drugs. Biological macromolecules like serum albumins have a crucial role in the design and biological safety assessments of drugs. Amikacin as an example of the most used semi-synthetic aminoglycosides antibiotics, is included in the crucial drug list of the World Health Organization. This in vitro study aimed to explore Amikacin's binding specification with bovine serum albumin (BSA) through a computational and experimental approach. According to fluorescence spectra, it was declared that the innate fluorescence emission of serum albumin was quenched by the Amikacin via a static quenching mechanism. Meantime, the binding constants and thermodynamic parameters of Amikacin with BSA were calculated at various temperatures. Based on this calculation, negative ΔG°, ΔH°, and ΔS° values showed that spontaneous binding process occurred via hydrogen bond and van der Waals interaction. Additionally, based on UV–vis absorption, Amikacin leads to conformational modifications of BSA with the construction of a ground state complex, which may affect the physiological functions of this serum albumin. The results of FTIR spectroscopy showed that the binding of amikacin led in the change of BSA secondary structure and confirmed the possibility of changing the physiological functions of BSA. It is anticipated that this research will supply significant insight into the pharmacological properties of Amikacin and its related effects on human health in vivo.

Low-moisture extruded mung bean protein isolate and wheat gluten: Structure, techno-functional characteristics and establishment of rehydration kinetics of the products

Low-moisture extruded products provide excellent shelf-life and stability, making the exploration of new formulations with various proteins highly valuable. This study prepared and compared products with different ratios of mung bean protein isolate (MPI) to wheat gluten (WG) (10:0, 9:1, 8:2, 7:3, 6:4, and 5:5). Results showed significant improvements (P < 0.05) in L∗-value and product quality with increasing WG ratios, peaking at an MPI/WG ratio of 7:3. Increased WG also enhanced protein cross-linking aggregation, as evidenced by changes in particle size, secondary structure, and fluorescence spectra. However, excessive WG (MPI/WG = 6:4) adversely affected product structure and techno-functional characteristics. Rehydration kinetics were best described by the Peleg model, with rehydrated products showing decreased hardness (291.25 N) and chewiness (227.45 N), but increased tensile resistance (P < 0.05). Eleven quadratic polynomial equations were developed to predict color and techno-functional characteristics based on WG percentage (0–50%). The findings provide a theoretical basis for expanding the application of MPI in new low-moisture extruded products.

Impact of ultrasonic and heat treatments on the physicochemical properties and rennet-induced coagulation characteristics of milk from various species

This study investigates the effects of heat and ultrasonic treatments on the physicochemical parameters and rennet-induced coagulation properties of milk from a variety of species, including cow, goat, buffalo, and donkey. Milk samples were subjected to heat treatments at different temperatures (65 °C, 80 °C, 90 °C, 100 °C) and ultrasonic treatment at varying power levels (200 W, 400 W, 600 W, 800 W, 1000 W). The results revealed that changes in turbidity, particle size, zeta potential, secondary structure, and surface hydrophobicity were altered by both ultrasonic and heat treatments, as well as the kind of milk. Ultrasonic treatment of cow milk decreased α-helix content while increasing β-turn content. Under similar ultrasonic treatment, goat milk showed a considerable increase in β-sheet content, whereas β-turn and random coil contents decreased compared to control samples. Notably, the water-holding capacity of gels formed from all four types of milk increased significantly with the intensity of ultrasonic and heat treatments. The hardness of buffalo milk gels increased significantly after ultrasonic and thermal treatments, ranging from 63 °C for 30 min to 90 °C for 15 min, but the hardness of cow and goat milk gels increased in varying degrees compared to their control samples. Furthermore, gels from cow and goat milk had higher storage modulus (G’) and loss modulus (G’’) than those from buffalo and donkey milk, and changes in G’ and G’’ from the examined milk were altered by ultrasonic and heat treatments. These findings offer important insights into refining milk processing procedures to improve dairy product quality and usefulness.

Exploring the Kinetics and Thermodynamics of a Novel Histidine Ammonia-Lyase from Geobacillus kaustophilus.

Histidine ammonia-lyase (HAL) plays a pivotal role in the non-oxidative deamination of L-histidine to produce trans-urocanic, a crucial process in amino acid metabolism. This study examines the cloning, purification, and biochemical characterization of a novel HAL from Geobacillus kaustophilus (GkHAL) and eight active site mutants to assess their effects on substrate binding, catalysis, thermostability, and secondary structure. The GkHAL enzyme was successfully overexpressed and purified to homogeneity. Its primary sequence displayed 40.7% to 43.7% similarity with other known HALs and shared the same oligomeric structure in solution. Kinetic assays showed that GkHAL has optimal activity at 85 °C and pH 8.5, with high thermal stability even after preincubation at high temperatures. Mutations at Y52, H82, N194, and E411 resulted in a complete loss of catalytic activity, underscoring their essential role in enzyme function, while mutations at residues Q274, R280, and F325 did not abolish activity but did reduce catalytic efficiency. Notably, mutants R280K and F325Y displayed novel activity with L-histidinamide, expanding the substrate specificity of HAL enzymes. Circular dichroism (CD) analysis showed minor secondary structure changes in the mutants but no significant effect on global GkHAL folding. These findings suggest that GkHAL could be a promising candidate for potential biotechnological applications.

Enhanced stability and catalytic performance of immobilized phospholipase D on chitosan-encapsulated magnetic nanoparticles using oxidized dextran and glutaraldehyde as cross-linkers

Phospholipase D (PLD) is essential for the bioconversion of phosphatidylcholine (PC) to phosphatidylserine (PS), a process valuable in functional food and medicine. This study explores the stability and catalytic properties of PLD immobilized on chitosan-encapsulated magnetic nanoparticles (CMNPs), utilizing oxidized dextran (DX) and glutaraldehyde (Glu) as cross-linkers. The cross-linker concentration and immobilization time were optimized to assess their effects on PLD catalytic performance. PLD immobilized on CMNPs with DX (DX-CMNPs-PLD) exhibited optimal activity at pH 8.0 and 30 °C, retaining over 40 % activity after 14 cycles, while Glu-cross-linked PLD (Glu-CMNPs-PLD) retained approximately 65 %. DX-CMNPs-PLD demonstrated superior pH, temperature, and operational stability compared to free PLD. Additionally, the immobilized PLD was characterized using transmission electron microscopy, X-ray diffraction, and Fourier-transform infrared spectroscopy. Kinetics parameters (Vmax and Km) of the immobilized PLD were also studied with free PLD serving as a control. Conformational analyses indicated a significant change in PLD's secondary structure, particularly in β-sheet content, which likely contributed to the enhanced stability and activity. These findings suggest a promising approach for PLD immobilization on CMNPs, with notable implications for biotechnological applications.

Mechanically rigid metallopeptide nanostructures achieved by highly efficient folding

AbstractNatural proteins must fold into complex three-dimensional structures to achieve excellent mechanical properties vital for biological functions, but this has proven to be exceptionally difficult to control in synthetic systems. As such, the long-standing issue of low mechanical rigidity and stability induced by misfolding constrains the physical and chemical properties of self-assembling peptide materials. Here we introduce a mixed-chirality strategy that enhances folding efficiency in topologically interlocked metallopeptide nanostructures. The orderly entanglement of heterochiral peptide-derived linkers can fold into a compact three-dimensional catenane. These folding-mediated secondary structural changes not only generate biomimetic binding pockets derived from individual peptide strands but also result in strong chiral amplification by the tight interlocking manner. Notably, this strategic ‘chirality mutation’ alters their arrangement into tertiary structures and is pivotal in achieving exceptional mechanical rigidity observed in the metallopeptide crystals, which exhibit a Young’s modulus of 157.6 GPa, approximately tenfold higher than the most rigid proteinaceous materials in nature. This unusual nature is reflected in enhanced peptide-binding properties and heightened antimicrobial activities relative to its unfolded counterpart.

The effects of Cannabis sativa and cannabinoids on the inhibition of pancreatic lipase – An enzyme involved in obesity

IntroductionObesity is a chronic noncommunicable disease characterized by excessive body fat that can have negative health consequences. Obesity is a complex disease caused by a combination of genetic, environmental, and lifestyle factors. It is characterized by a discrepancy between caloric intake and expenditure. Obesity increases the risk of acquiring major chronic diseases, including heart disease, stroke, cancer, and Type 2 diabetes mellitus (T2DM). Currently, the inhibition of pancreatic lipases (PL) is a promising pharmacological therapy for obesity and weight management. In this study, the inhibition of pancreatic lipase by Cannabis sativa (C. sativa) plant extract and cannabinoids was investigated. MethodsThe inhibitory effect was assessed using p-nitrophenyl butyrate (pNPB), and the results were obtained by calculating the percentage relative activity and assessed using one-way analysis of variance (ANOVA). Kinetic studies and spectroscopy techniques were used to evaluate the mode of inhibition. Diet-induced; and diabetic rat models were studied to evaluate the direct effects of C. sativa extract on PL activity. ResultsKinetic analyses showed that the plant extracts inhibited pancreatic lipase, with tetrahydrocannabinol (THC) and cannabinol (CBN) being the potential cause of the inhibition noted for the C. sativa plant extract. CBN and THC inhibited the pancreatic lipase activity in a competitive manner, with the lowest residual enzyme activity of 52 % observed at a 10 μg/mL concentration of CBN and 39 % inhibition at a 25 μg/mL concentration of THC. Circular dichroism (CD) spectroscopy revealed that the inhibitors caused a change in the enzyme's secondary structure. At low concentrations, THC showed potential for synergistic inhibition with orlistat. C.sativa treatment in an in vivo rat model confirmed its inhibitory effects on pancreatic lipase activity. ConclusionThe findings in this study provided insight into the use of cannabinoids as pancreatic lipase inhibitors and the possibility of using these compounds to develop new pharmacological treatments for obesity.

PH-induced interface protein structure changes to adjust the stability of tilapia protein isolate emulsion prepared by high-pressure homogenization

The pH is a crucial external factor affecting the structure and emulsification characteristics of proteins. The current study aimed to reveal the correlation between the secondary structure changes and tilapia protein isolate (TPI) emulsion stability under different pH (3.0–10.0) prepared by high-pressure homogenization. The results showed that TPI with significantly increased solubility and emulsifying properties when the pH keep away from the isoelectric point (pH 5.0). Meanwhile, TPI emulsions presented significantly enhanced stability (with decreased particle size, increased zeta potential, creaming index close to 0, and uniform dispersion of droplets) at pH 3.0 and 10.0. Interface-adsorbed protein mainly consists of a myosin-heavy chain and actin, and the secondary structure was significantly influenced by pH and high-pressure homogenization. The α-helix will be transformed into β-sheet and β-turn when pH is closer to pH 5.0. However, the high-pressure homogenization induced α-helix conversion to β-sheet. The correlation analysis revealed that emulsion stability is positively correlated with α-helix and negatively correlated with β-sheet. This work provides a deep insight into the correlation between secondary structure changes and the stability of TPI emulsion as affected by pH to offer an alternative way to enhance TPI emulsion stability.

Development of levamlodipine long-acting patches based on an ion-pair strategy: Investigation of the mechanism for reducing skin irritation

The aim of this study was to develop a long-acting transdermal patch of levamlodipine (LAM) using an ion-pair strategy to reduce the skin irritation induced by topical application of LAM and explore the mechanism underlying the improvement of skin irritation. The formulation was optimized through porcine in vitro transdermal experiments and rabbit in vivo skin irritation tests. The obtained formulation consisted of poly (2-Ethylhexyl acrylate-co-N-Vinyl-2-pyrrolidone-co-N-(2-Hydroxyethyl) acrylamide) (PENH) as the adhesive matrix, 13.00 % levamlodipine-sorbic acid ion-pair complex (LAM-SA) (w/w), and 10 % isopropyl myristate (IPM) (w/w), with a patch thickness of 70 μm, achieving an erythema index of 188 for rabbit skin and 117–187 for human skin (264 for rabbit skin and 110–260 for human skin in the absence of sorbic acid (SA)). In vivo rabbit and human skin erythema analysis and H&E staining verified that the optimized ion-pair patch effectively reduced skin irritation. Drug distribution experiments in the skin, ATR-FTIR, and molecular simulation were used to characterize the mechanism by which the ion-pair reduced skin irritation. Excessive accumulation of LAM in the epidermis induced secondary structural changes in keratin, resulting in skin barrier damage and inflammatory response. The formation of the LAM-SA ion pair altered physicochemical properties of LAM, reducing drug retention in the epidermis and, thereby, reducing skin irritation. This study demonstrated the potential of the ion-pair strategy to improve the safety of transdermal drug delivery system (TDDS) and provided a means for reducing skin irritation caused by the active pharmaceutical ingredient (API) itself.

Impact of pulsed light treatment on enzyme inactivation and quality attributes of whole white button mushroom (Agaricus bisporus) and its storage study

Whole white button mushrooms (WWBM) exhibit a limited shelf-life owing to the oxidative enzymatic browning. Inactivation of polyphenol oxidase-PPO and peroxidase-POD in WWBM and its kinetic behavior were studied using pulsed light(PL) treatment (0.13–1.11 J/cm2). The first-order kinetics explained PL-induced enzyme inactivation. Rate constants(k) for PPO and POD were 3.84 and 2.55cm2/J. FTIR-analysis revealed secondary-structural changes in partially-purified enzyme. PL-treatment retarded browning, retained phenolics and enhanced vitamin D2. PL-treatment at 1.11 J/cm2 rendered WWBM both microbially and enzymatically stable. The PL-treated WWBM's shelf-life at 4, 20, and 37 °C were 5, 3, and 1 day. At 4 °C, browning increased by 6.1 %; firmness decreased by 55.2 %, while PL-treated mushrooms retained 90.6 % phenolics, 78.9 % antioxidant capacity, and 64.2 % D2 after 5 days. Higher activation energy value confirmed phenolics were most sensitive during storage. PL-technology supports UN Sustainable Development Goals by reducing chemical use, lowering carbon-footprints, minimizing pollution, and enhancing shelf-life, promoting sustainable global trade.

Allosteric mechanism of synergistic effect in α- and β-amylase mixtures

Alpha-amylase and beta-amylase coexist as mixtures in industrial production, and the two amylases have active synergistic effects when they approach each other. These effects are due to enhanced enzyme binding affinity for the substrate and the rate of particle hydrolysis. Here, we report the allosteric mechanism of this synergistic effect in α- and β-amylase mixtures. The assay showed higher activity after mixing α- and β-amylase. Molecular docking showed that α- and β-amylase create a stable dual-enzyme complex with high binding energy, and that complex formation does not affect the exposure of respective active sites. β-Amylase is specifically bound to the B domain of α-amylase, and the dynamic plasticity of the B domain makes it move spatially, and this adjustment leads to a more open conformation in the active site of α-amylase. Because the enzymes binding make the complex more stable, the degree to which the relative activity of the dual-enzyme complex is inhibited is significantly reduced. After enzyme hydrolysis, the products maltose and glucose accumulate and produce competitive inhibition, which explains the relative activity decrease of the later-stage dual-enzyme cooperation. Structural characterization by FT-IR and CD spectroscopy did not reveal significant changes in respective secondary structures after enzyme binding.

Long-chain fatty acids block allergic reaction against lipid transfer protein Sola l 7 from tomato seeds.

Due to the benefits of tomato as an antioxidant and vitamin source, allergy to this vegetable food is a clinically concerning problem. Sola l 7, a class I lipid transfer protein found in tomato seeds, has been identified as an allergen linked to severe anaphylaxis. However, the role of lipid binding in Sola l 7-induced allergy remains unclear. Here, the three-dimensional structure of recombinant Sola l 7 (rSola l 7) has been elucidated using nuclear magnetic resonance spectroscopy (NMR). Its interaction with free fatty acids has been deeply studied; fluorescence emission spectroscopy revealed that different long-chain fatty acids interact with the protein, affecting the only tyrosine residue present in Sola l 7. On the contrary, no changes in the overall secondary structure were observed after the analysis of the circular dichroism spectra in the presence of fatty acids. Unsaturated oleic and linoleic fatty acids presented higher affinity and promoted more significant changes than saturated or short-chain fatty acids. 1H-15N HSQC NMR spectra allowed to determine the regions of the protein that were modified when rSola l 7 interacts with the fatty acids, suggesting epitope modification after the interaction. For corroboration, IgG and IgE binding to rSola l 7 were assessed in the presence of free fatty acids, revealing that both IgE and IgG binding were significantly lower than in their absence, suggesting a potential protective role of unsaturated fatty acids in tomato allergy.

Bio-spectroscopic analysis of corneal structural alterations in dry eye disease: A study of collagen, co-enzymes, lipids, and proteins with emphasis on phytotherapy intervention

Dry eye disease (DED) stands as a prevalent cause for ophthalmology consultations, securing the third position following refractive errors and cataracts. Moreover, the likelihood of experiencing DED escalates with advancing age. In this experimental study corneal tissue alterations due to DED were investigated over different periods by applying both infrared and synchronous fluorescence spectroscopy. The potential effects of instillation of pomegranate and green tea water extracts as green-friendly treatment modalities were also evaluated. The obtained results collectively indicate that DED affects the OH bearing constituents (collagen, glycosaminoglycans, and proteoglycans) of cornea leading to changes in protein secondary structure and the collagen fibrils. Additionally, enhanced dehydrated environment, and reduced energetic/metabolic state, as indicated by co-enzymes, was observed. Phyto-therapeutic administration can contain these alterations with enhanced energetic/metabolic state and increased hydration environment. In conclusion, instillation of green tea extract can protect/restore the collagen fibrils and its potential effects, in general, exceeds that of pomegranate extract.

Thermal-induced interactions between soy protein isolate and malondialdehyde: Effects on protein digestibility, structure, and formation of advanced lipoxidation end products

Thermally processed lipid- and protein-rich foods have sparked widespread concern since they may degrade food nutrition and even risk food safety. This study investigated soy protein isolate (SPI) alterations of digestibility and structure, as well as the formation of potentially hazardous chemicals, i.e., advanced lipoxidation end products (ALEs), after interacting with malondialdehyde (MDA, a lipid oxidation product) under high-temperature cooking conditions (100–180 °C, up to 60 min). In-vitro protein digestion of the SPI-MDA mixtures suggested that their room-temperature interactions damaged SPI digestibility, and increasing the temperature and the duration of the thermal treatment exacerbated the adverse effects. Protein oxidation, covalent aggregation of subunits, and changes in secondary and tertiary structures were revealed using thiol quantification, gel electrophoresis, fluorescence spectroscopy, and circular dichroism (CD) spectra, which could explain reduced protein digestibility. High-resolution mass spectrometry (HRMS) identified seven non-crosslinked ALEs and two crosslinked ALEs. Increased MDA concentrations promoted the generation of ALEs. Moreover, the acrolein-derived ALEs with reactive carbonyl groups were prone to further reacting into crosslinked ALEs, potentially responsible for the subunit aggregation.

Chikungunya Virus RNA Secondary Structures Impact Defective Viral Genome Production.

Chikungunya virus (CHIKV) is a mosquito-borne RNA virus that poses an emerging threat to humans. In a manner similar to other RNA viruses, CHIKV encodes an error-prone RNA polymerase which, in addition to producing full-length genomes, gives rise to truncated, non-functional genomes, which have been coined defective viral genomes (DVGs). DVGs have been intensively studied in the context of therapy, as they can inhibit viral replication and dissemination in their hosts. In this work, we interrogate the influence of viral RNA secondary structures on the production of CHIKV DVGs. We experimentally map RNA secondary structures of the CHIKV genome using selective 2'-hydroxyl acylation analyzed by primer extension and mutational profiling (SHAPE-MaP), which couples chemical labelling with next-generation sequencing. We correlate the inferred secondary structure with preferred deletion sites of CHIKV DVGs. We document an increased probability of DVG generation with truncations at unpaired nucleotides within the secondary structure. We then generated a CHIKV mutant bearing synonymous changes at the nucleotide level to disrupt the existing RNA secondary structure (CHIKV-D2S). We show that CHIKV-D2S presents altered DVG generation compared to wild-type virus, correlating with the change in RNA secondary structure obtained by SHAPE-MaP. Our work thus demonstrates that RNA secondary structure impacts CHIKV DVG production during replication.

Engineered polymeric excipients for enhancing the stability of protein biologics: Poly(N-isopropylacrylamide)-poly(ethylene glycol) (PNIPAM-PEG) block copolymers

Protein therapeutics, particularly antibodies, depend on maintaining their native structures for optimal function. Hydrophobic interfaces, such as the air–water interface, can trigger protein aggregation and denaturation. While completely avoiding such interfacial exposures during manufacturing and storage is impractical, minimizing them is crucial for enhancing protein drug stability and extending shelf life. In the biologics industry, surfactants like polysorbates are commonly used as additives (excipients) to mitigate these undesirable interfacial exposures. However, polysorbates, the most prevalent choice, have recognized limitations in terms of polydispersity, purity, and stability, prompting the exploration of alternative excipients. The present study identifies poly(N-isopropylacrylamide)-poly(ethylene glycol) (PNIPAM-PEG) block copolymers as a promising alternative to polysorbates. Due to its stronger affinity for the air–water interface, PNIPAM-PEG significantly outperforms polysorbates in enhancing protein stability. This claim is supported by results from multiple tests. Accelerated dynamic light scattering (DLS) experiments demonstrate PNIPAM-PEG’s exceptional efficacy in preserving IgG stability against surface-induced aggregation, surpassing conventional polysorbate excipients (Tween 80 and Tween 20) under high-temperature conditions. Additionally, circular dichroism (CD) spectroscopy results reveal conformational alterations associated with aggregation, with PNIPAM-PEG consistently demonstrates a greater protective effect by mitigating negative shifts at λ ≅ 220 nm, indicative of changes in secondary structure. Overall, this study positions PNIPAM-PEG as a promising excipient for antibody therapeutics, facilitating the development of more stable and effective biopharmaceuticals.

Insight into the self-assembly process, microrheological properties and microstructure of acid-soluble collagen from lamb skin under different environmental conditions

The self-assembly behavior of collagen under the regulation of the external environment is important for the construction of collagen-based materials. In this study, the effects of temperature, pH, and ion concentration on the self-assembly property, microrheology, and microstructure of acid-soluble collagen (ASC) extracted from lamb skin were investigated. The kinetics of ASC self-assembly under different environmental conditions was captured by monitoring the turbidity over time. The results showed that the ASC assembly process was accelerated by temperature increase in the range of 25–37 °C, and the self-assembly was unfavorable when the temperature exceeded 37 °C. The pH closed to the isoelectric point of ASC was conducive to assembly, and appropriate ion concentration increased the assembly rate of ASC. The microrheology reflected the influence of environmental conditions on the assembly process through the changes in the elastic and solid-liquid transition points of the solution system, and the results were consistent with the conventional turbidimetry method. Scanning electron microscopy showed that ASC formed fibril structures with different diameters in different environments. In addition, infrared spectroscopy showed the changes of secondary structure of ASC during the assembly process. The amino acid composition, UV spectrum, secondary structure, and morphology of ASC were characterized. The above results provide a foundation for the construction of controllable ASC self-assemblies from lambskins.

Adaptability to the environment of protease by secondary structure changes and application to enzyme-selective hydrolysis

The reactions involving enzymes are significantly influenced by various environmental factors. Clarity of how the activity and structure of proteases impact their function is crucial for more efficient application of enzymes as a tool. The impact of temperature, pH, and ionic strength on changes in protease activity, secondary structure, and protein conformation during enzymatic hydrolysis were investigated in this study. The enzymatic activity and secondary structure of acid-base protease were found to undergo significant modifications under different physical conditions, as demonstrated by UV spectrophotometry and FTIR spectroscopy analysis. Specifically, variations in α-helix and β-fold content were observed to correlate with changes in enzyme activity. Molecular simulation analysis revealed that physical conditions have varying effects on the protease, particularly influencing enzyme activity and secondary structure. Evaluation of the proteases indicated alterations in both enzyme activity and structure. This treatment selectively hydrolyzed β-lactoglobulin and reduced sensitization. These findings offer novel perspectives on the functionalities and regulatory mechanisms of proteases, as well as potential industrial applications.

Molecular characterisation of interactions between 11S glycinin and hexanal – An off flavour compound

This work examines the binding characteristics between the 11S glycinin from soy flour with the aldehyde compound hexanal, at near neutral pH and ambient temperature. FTIR and CD spectroscopy determined that secondary structure changes were evident upon complexation with observed increases in α-helical and unordered structures along with reductions in β-sheets and β-turn structures upon complexation. UV–Vis and MALDI-TOF-MS analysis confirmed the physical nature of hexanal stabilisation, with in silico analysis and fluorescence spectroscopy revealed a low to medium binding strength with a KA range of 3.1 × 102 to 3.1 × 104 M−1 along with a 1:1 binding stoichiometry between the compound and 11S. Molecular docking simulations determined the binding location is situated between chain A and B of the trimer and is largely stabilised by pi-alkyl, hydrogen and van der Waals forces. Findings from this study will provide increased knowledge in the field of flavour compound and protein interactions thus advancing our knowledge of flavour binding and release in plant-based beverage formulations.