Sulfhydryl Research Articles

Unravelling structural insights into ligand-induced photoluminescence mechanisms of sulfur dots.

Sulfur dots (S-QDs) hold promise as a new category of metal-free, luminescent nanomaterials, yet their practical application faces challenges primarily due to a limited understanding of their structure and its impact on their optical properties. Herein, by employing a spectrum of aliphatic and aromatic ligands, we identify the surface structure and composition of S-QDs while delineating the pivotal role of ligands in inducing photoluminescence. Thiol-functionalized ligands, such as 4-mercapto benzoic acid and glutathione, notably promote the formation of both green and blue luminescent S-QDs, boosting a high quantum yield of up to 56%. Further investigation on the synthesis of S-QDs with 4-mercapto benzoic acid unveils the dual role of H2O2: etching sulfur powder and oxidizing the -SH group to -SO2H. These oxidized ligands passivate the S-QD surface through hydrogen bonding. Electrospray ionization mass spectrometry analysis unveils the presence of distinct sulfur species such as [S4(C6H5SO2H)4(H2O)2H]+ and [S6(C6H5SO2H)6(H2O)3H]+, while XPS analysis confirms the existence of zerovalent sulfur and oxidized sulfur species including SO32- and SO42-. Further detailed spectroscopic examination demonstrates that S-QDs predominantly exist as aggregated entities, with the emission wavelength correlating with the degree of aggregation. The absence of photoluminescence in aggregations devoid of ligands underscores the critical role of ligands in the photoluminescence genesis of S-QDs.

Defect-Engineered MOF-801/Sodium Alginate Aerogel Beads for Boosting Adsorption of Pb(II).

Metal-organic frameworks (MOFs) are attractive adsorbents for heavy metal capture due to their superior stability, easy modification, and adjustable pore size. However, their inherent microporous structure poses challenges in achieving a higher adsorption capacity. Defect engineering is considered a simple method to create hierarchical MOFs with larger pores. Here, we employed l-aspartic acid as a mixed linker to bind Zr4+ clusters in competition with fumaric acid of MOF-801 to create defects, and the pore size was increased from 4.66 to 15.65 nm. Mercaptosuccinic acid was subsequently used as a postexchange ligand to graft the resultant MOF-801 by acid-ammonia condensation to further expand the pore size to 22.73 nm. Notably, the -NH2, -COOH, and -SH groups contributed by these two ligands increased the adsorption sites for Pb(II). The obtained defective MOF-801 with larger pores was thereafter loaded onto sodium alginate to form aerogel beads as adsorbents, and an adsorption capacity of 375.48 mg/g for Pb(II) was achieved, which is ∼51 times that of pristine MOF-801. The aerogel beads also exhibited outstanding reusability with a removal efficiency of ∼90.23% after 5 cycles of use. The adsorption mechanism of Pb(II) included ion-exchange interaction, as well as chelation interactions of Pb-O, Pb-NH2, and Pb-S. The versatile combination of defect engineering and composite beads provides novel inspirations for MOF modification for boosting heavy metal adsorption.

Spray dried protein concentrates from white button and oyster mushrooms produced by ultrasound-assisted alkaline extraction and isoelectric precipitation.

In the present study, the optimization of ultrasound-assisted alkaline extraction (UAAE) and isoelectric precipitation (IEP) was applied to white button (WBM) and oyster (OYM) mushroom flours to produce functional spray dried mushroom protein concentrates. Solid-to-liquid ratio (5-15% w/v), ultrasound power (0-900 W) and type of acid [HCl or acetic acid (AcOH)] were evaluated for their effect on the extraction and protein yields from mushroom flours submitted to UAAE-IEP protein extraction. Prioritized conditions with maximized protein yield (5% w/v, 900 W, AcOH, for WBM; 5% w/v, 900 W, HCl for OYM) were used to produce spray dried protein concentrates from white button (WBM-PC) and oyster (OYM-PC) mushrooms with high solids recovery (62.3-65.8%). WBM-PC and OYM-PC had high protein content (5.19-5.81 g kg-1), in addition to remarkable foaming capacity (82.5-235.0%) and foam stability (7.0-162.5%), as well as antioxidant phenolics. Highly pH-dependent behavior was observed for solubility (> 90%, at pH 10) and emulsifying properties (emulsification activity index: > 50 m2 g-1, emulsion stability index: > 65%, at pH 10). UAAE-IEP followed by spray drying increased surface hydrophobicity and free sulfhydryl groups by up to 196.5% and 117.5%, respectively, which improved oil holding capacity (359.9-421.0%) and least gelation concentration (6.0-8.0%) of spray dried mushroom protein concentrates. Overall, the present study showed that optimized UAAE-IEP coupled with spray drying is an efficient strategy to produce novel mushroom protein concentrates with enhanced functional attributes for multiple food applications. © 2024 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Enhanced proton transfer in proton exchange membrane fuel cells via novel nanocomposite membrane incorporating (3-Mercaptopropyl)trimethoxysilane-graphene oxide and basic amino ligands: A synergistic acid-base approach

A nanocomposite polymer electrolyte membrane (MGC) was synthesized by blending MPTS (HS(CH2)3Si(OCH3)3) and graphene oxide (GO) nanosheets at varying weight ratios (91.5 to 9.5, respectively) for proton exchange membrane fuel cell (PEMFC) applications. By covalently modifying the GO support with MPTS through a silane modification process, the acidic MGC matrix is formed. The blending process, with ultrasonication and vigorous stirring, initiates two significant reactions in reactive MPTS: first, the conversion of methoxy groups (Si–OCH3) to silanols (Si-OH); second, the potential condensation of silanol groups to form siloxane bonds (Si–O–Si), followed by oxidation of sulfhydryl groups to sulfonic acid (-SO3H) groups by hydrogen peroxide. The resulting -SO3H groups connected to the siloxane network (Si-O-Si) adjacent to the GO. Two basic amino ligands – dopamine, and aminopropyl trimethoxysilane (APTES) were incorporated into the MGC matrix to establish acid-base pairs, promoting swift proton transport. These additions, at weight ratios of (0.5, 1, 2, and 7) wt%, synergistically contributed to the MGC matrix, forming effective proton channels. Various analytical techniques, including field emission-scanning electron microscopy (FE-SEM), atomic force microscopy (AFM), Raman spectroscopy, X-ray diffraction (XRD), and electrochemical impedance spectroscopy (EIS), were employed to assess the impact of functionalized basic amino groups on the MGC matrix. Notably, the MGC membrane containing 2 wt% of PDA as basic amino ligands exhibited optimal performance, showcasing an in-plane proton conductivity of 115.10 mS cm-1 under fully hydrated conditions at 90 °C, and the maximum power density (MPD) of 170.74 mW cm-2 with a current density of 1051.46 mA cm-2 at 60 °C and 100 % relative humidity (RH). In contrast, the MGC membrane, devoid of amino ligands, exhibited a comparatively low in-plane conductivity of 44.32 mS cm⁻¹ under identical conditions at 90 °C. Additionally, it recorded the MPD of 48.5 mW cm⁻², corresponding to a current density of 313.25 mA cm⁻² at 60 °C and 100 % RH. This study introduces an innovative approach for developing high-performance nanocomposite membranes for PEMs and related applications.

Growth and stabilization of high-concentration metallic 1T-phase MoS2 as a high-performance l-cysteine electrochemical sensor by electron injection engineering

MoS2 is an attractive electrochemical sensing electrode material for the detection of biomolecule because of its adjustable band gap, large surface area, and unique electrochemical characteristics. However, the poor catalytic and electrical conductivity of the semiconductor 2H-phase MoS2 and the phenomenon of restacking during electrochemical use hinder its sensing application. Herein, electron injection engineering is employed for the formation and stabilization of high-concentration metal 1T-phase MoS2 nanosheets on ionic liquid-functionalized carbon nanotubes (CNTs-IL). The increase of MoS2 concentration in the metal 1T phase significantly accelerates electron transfer kinetics. The in-situ growth method results in a tightly interfaced coupling effect, thereby substantially shortening the electron transport pathway and decreasing interfacial transmission impedance. In addition, the sulfur defects on the surface of MoS2 as the anchoring point of sulfhydryl groups promote the enrichment of thiols on the surface of the material and reduce the reaction barrier for sulfhydryl oxidation, thereby improving the catalytic performance. The prepared CNTs-IL-MoS2/SPE biosensor shows excellent l-cysteine sensing performance by electrochemical detection.

Elucidating the gas cell stabilization mechanism of buckwheat-wheat steamed bread induced by transglutaminase: A focus on the foaming and air-water interfacial properties of dough liquor

This work investigated the improvement mechanism of transglutaminase (TG) on the buckwheat-wheat steamed bread qualities from the view of protein structural, foaming and air-water interfacial properties of dough liquor (DL). TG reduced the hardness, ameliorated the crumb structure and enhanced the specific volume of buckwheat-wheat steamed bread by up to 9.0%. The DL yield, protein and water recovery were notably increased as the TG concentration increased to 0.2 U/g, then decreased with the higher TG concentrations. The low TG concentrations (<0.2 U/g) increased the protein molecular weight, and decreased the protein hydrophobicity, free amino and sulfhydryl groups content of DL. The high TG concentrations (>0.2 U/g) tended to induce both protein intermolecular and intramolecular cross-linking, which reduced the average particle size from 9.6 to 4.3 μm as the TG concentration increased from 0.2 to 1.0 U/g. TG markedly improved the foaming capacity and foam stability, decreased the surface tension and strengthened the interfacial films by enhancing the viscoelasticity modulus of buckwheat-wheat DL interfacial protein films. The topography of the air-water interfacial layer revealed that the TG-induced protein cross-linking led to a more homogeneous and denser topographical structure. This work will provide fresh insight into the effects of TG-induced protein cross-linking on gas cell stabilization and relate them to the buckwheat-wheat steamed bread quality properties.

Microwave-assisted, sulfhydryl-modified β-cyclodextrin-silymarin inclusion complex: A diverse approach to improve oral drug bioavailability via enhanced mucoadhesion and permeation

The current study aimed to generate a sulfhydryl-modified β-cyclodextrin-silymarin complex (sulfhydryl-modified β-CD-SMN complex) and to evaluate the enchantment in solubility, permeability, and bioavailability of a model BCS Class IV drug silymarin (SMN). For this purpose, sulfhydryl-modified β-CD was synthesized by replacing all primary and secondary –OH groups at the β-CD backbone with sulfhydryl groups via a novel microwave-assisted technique. Afterward, sulfhydryl-modified β-CD was complexed with silymarin and characterized by FTIR and 1H NMR spectroscopy. Moreover, no. of sulfhydryl groups and their oxidative stability, solubility, safety, mucoadhesion, release, diffusion, and rheological studies were performed. Furthermore, in-vivo studies were conducted to confirm enhanced pharmacokinetic properties of silymarin. Sulfhydryl-modified β-CD showed 8291 ± 418 μmol/g sulfhydryl groups that were prone to oxidation at pH ≥ 5, however, most of the sulfhydryl groups were found stable at pH 4 having a pKa value of 8.3. Modified β-CD oligomer showed improved solubility of SMN, significantly enhanced drug transport across goat intestinal mucosa, 78-fold improved mucoadhesion, improved drug dissolution and 4.4-fold enhanced dynamic viscosity. No toxic effects were reported to Caco-2 cells at 0.5 % (m/v) concentration of sulfhydryl-modified β-CD for 24 h. The apparent permeability coefficient (Papp) of SMN was 6.9-fold enhanced on goat intestinal mucosa. Moreover, in-vivo studies confirmed a significantly enhanced oral bioavailability of SMN due to combination with sulfhydryl-modified β-CD. Based on these findings, the sulfhydryl-modified β-CD-silymarin inclusion complex can be a promising technique to enhance the bioavailability of BCS Class IV drugs via enhanced solubility, mucoadhesion, and permeability triple action.

Quercetin Prevents Hypertension in Dahl Salt-sensitive Rats F ed a High-salt Diet Through Balancing Endothelial Nitric Oxide Synthase and Sirtuin 1.

A high-salt diet is a leading dietary risk factor for elevated blood pressure and cardiovascular disease. Quercetin reportedly exhibits cardioprotective and antihypertensive therapeutic effects. The objective of this study is to examine the effect of quercetin on high-salt dietinduced elevated blood pressure in Dahl salt-sensitive (SS) rats and determine the underlying molecular mechanism. Rats of the Dahl SS and control SS-13 BN strains were separated into five groups, SS-13 BN rats fed a low-salt diet (BL group), SS-13 BN rats fed a high-salt diet (BH group), Dahl SS rats fed a low-salt diet (SL group), Dahl SS rats fed a high-salt diet (SH group), and SH rats treated with quercetin (SHQ group). Blood pressure was checked three weeks into the course of treatment, and biochemical markers in the urine and serum were examined. Additionally, western blot was done to evaluate the sirtuin 1 (SIRT1) and endothelial nitric oxide synthase (eNOS) expression levels. Immunohistochemical analysis was performed to verify SIRT1 levels. We demonstrated that a high-salt diet elevated blood pressure in both SS-13 BN and Dahl SS rats, and quercetin supplementation alleviated the altered blood pressure. Compared with the SH group, quercetin significantly elevated the protein expression of SIRT1 and eNOS. Immunohistochemistry results further confirmed that quercetin could improve the protein expression of SIRT1. Quercetin reduced blood pressure by enhancing the expression of SIRT1 and eNOS in Dahl SS rats fed a high-salt diet.

Colorimetric and smartphone-visual detection of biothiols in human serum and red wine based on POD-like activity of Fe-CDs

An efficient mimic peroxidase platform of Fe-CDs/GOx based on the hybrid cascade system to produce in-situ H2O2 for the visualized detection of glucose has been also developed in our group. Herein, the [Fe-CDs + H2O2 + TMB] system was further performed to detect the biothiols (GSH, Cys, and Hcy) as representatives of the –SH group. The result exhibits that the higher concentration of biothiols can cause the absorbance signal at 652 nm to decrease, and the deep-blue color of oxTMB turns green until it is faded in 20 min by the naked eye. The colorimetric method showed the LOD are 0.54, 0.29, and 1.41 μM for GSH, Cys, and Hcy respectively, without interference in the presence of other amino acids. The smartphone-based and paper-based determination display good respect for biothiols in human serum or GSH in red wine. These simple and convenient strategies provide potential applications in the fields of clinical diagnosis and foods.

Ultrasound treatment improved gelling and emulsifying properties of myofibrillar proteins from Antarctic krill (Euphausia superba)

Antarctic krill is a promising source of marine proteins with abundant biomass and excellent nutritional profile, but has poor technological properties. Ultrasonic treatment at power levels of 0, 100, 200, 300, 400 and 500 W was applied to improve the technological properties of Antarctic krill meat, and the changes in physicochemical properties of myofibrillar proteins (MPs) were investigated. The results indicated that proper ultrasonic treatment significantly improved the gelling properties of Antarctic krill meat, in terms of a more uniform and stable gel texture and better water holding capacity, which were related to better cross-linking of MPs. Ultrasonic treatment promoted the conversion of MPs’ secondary structures from α-helix and random coil to β-sheet and β-turn, thereby making the molecular structure soft and loose. In addition, at tertiary structure level, ultrasonic treatment exposed the hydrophobic groups and sulfhydryl groups within MPs, thereby improving the emulsifying properties by changing the intermolecular interactions and interface properties. Furthermore, the particle size of MPs decreased and exhibited a more uniform distribution, aligning with the enhanced interactions observed between MPs and oil. These results provide an insight into the efficient development of Antarctic krill by elucidating how the ultrasonic treatment improves the gelling and emulsifying properties based on structure modulation of myofibrillar proteins.

Exploration of the regulatory mechanism of pulsed electric field on the aggregation behavior of soybean protein isolates

As an emerging protein modification technology, pulsed electric field (PEF) technology in modifying soybean protein isolates (SPI) suffers from unclear mechanism and controversial changes in aggregation structure. To address these issues, the effects of PEF treatment with different electric field intensities (5–30 kV/cm) on the aggregation behavior and spatial structure of SPI were investigated. The results showed that the SPI aggregates exhibited a trend of depolymerization followed by reaggregation with the increase of PEF intensity. At 10 and 15 kV/cm, the polarization effect of PEF induced the unfolding of the tertiary structure of SPI, leading to the enhancement of the electronegativity of the side chains, and the electrostatic repulsive force between the molecules promoted the depolymerization of SPI aggregates. However, with the withdrawal of PEF, the structure of the SPI was partially reversible, resulting in a limited depolymerization effect. When the PEF intensity reached 20 kV/cm and above, SPI underwentstructure unfolding, subunit dissociation, continuous exposure of hydrophobic groups and sulfhydryl groups, and ultimately reaggregation mediated by hydrophobic interactions and disulfide bonding, resulting in the formation of high molecular weight soluble and insoluble protein aggregates. In addition, the formation of free radicals under strong electric fields (≥20 kV/cm) accelerated the oxidation of SPI and promoted the rapid formation of disulfide bonds. This study provides a theoretical basis for the targeted regulation of SPI aggregation structure by PEF.

The adsorption of cesium by sulfonic acid functionalized hollow mesoporous silica microspheres

With the development of nuclear industry, radioactive elements such as 137Cs put a threat on the water environment. It is a challenging task to remove the Cs+ in the nuclear wastewater. In the current study, we prepared a new Cs+-adsorbing material by introducing sulfhydryl group onto the surface of hollow mesoporous silica microspheres, then oxidizing the sulfhydryl group to sulfonic acid group. The obtained HMSS-SO3H material had an excellent adsorption capacity for Cs+ in the aqueous solution, with an adsorption capacity of 51.53 mg g−1 in 30 min. Characterization approaches, such as FT-IR and EDS, were used to confirm the result of modification. Adsorption experiments were carried out under. The influence of various parameters on the adsorption process was investigated under the conditions of changing pH, temperature, and time. The effect of competitive ions was also explored. The results indicated that the adsorption process followed the pseudo-second-order model and the main adsorption mechanisms are electrostatic interaction and coordination. The material had a best adsorption performance at a neutral pH. The adsorption process could well-fit the Langmuir's model, with a theoretical maximum adsorption capacity of 81.31 mg g−1. And the adsorption capacity was slightly affected by competing ions such as Mg2+ and Ca2+. The results indicate that the HMSS-SO3H prepared in this study is a promising adsorbent for Cs+, with the advantages of high adsorption capacity, fast adsorption rate and high selectivity.

Comparative analysis of the qualities of traditional and sous-vide marinated duck drumsticks.

The sous-vide technique is increasingly used to improve the quality of poultry meat; the study aimed to compare the quality of traditional and sous-vide marinated (SVM) duck drumsticks by analyzing the sensory-related, nutritional, storage-related, and in vitro digestive-related quality of duck meat. The results showed that the sensory quality scores of color, odor, and appearance, L* and a* values of duck drumsticks in SVM group were significantly increased compared with the traditional marinated (TM) group (t-test, p<0.05, the same below), and the b* values on the outside and inside of duck drumsticks were decreased by 22.47% and 38.04%, respectively. Compared with TM group, hardness, springiness, chewiness, adhesion, cohesion, and resilience of duck drumsticks in SVM group decreased by 43.32%, 29.52%, 65.08%, 62.35%, 20.23%, and 30.33%, respectively. The moisture content and total fat content of duck drumsticks in SVM group were significantly higher than those in TM group (p<0.05), and the protein loss, total volatile basic nitrogen, and thiobarbituric acid reactive substances values were decreased by 61.4%, 25.86%, and 20.45%, respectively. The results of in vitro digestion experiments showed that the content of free sulfhydryl groups of duck drumsticks in SVM group was significantly increased (p<0.05), and the contents of Schiff base and carbonyl groups were significantly decreased compared with the TM group (p<0.05). In conclusion, the SVM technology could significantly improve the sensory-related qualities, reduce the loss of nutrients, and improve the storage-related qualities of duck drumsticks. This study provided theoretical reference for the high-value application of SVM technology in duck meat.

Injectable self-healing alginate/PEG hydrogels cross-linked via thiol-Michael addition bonds for hemostasis and wound healing

In this study, an alginate/PEG hydrogel was developed via a thiol-Michael addition reaction between oxidized quinone of catechols on dopamine-grafted sodium alginate (SA-DA) and sulfhydryl groups of 4-arm polyethylene glycol tetra-thiol (4-arm PEG-SH) under mildly basic conditions. Through the formation of thiol-terminated catechol groups, the accompanying oxidized catechols are reduced, significantly strengthening the internal network structure of the hydrogel and improving tissue adhesion. Meanwhile, the hydrogels have excellent self-healing properties due to the dynamic non-covalent bonds between the groups. Adjustment of hydrogel properties by varying the mass ratio of two hydrogel precursors. Due to the high content of thiol-terminated catechol groups, the Gel 3 exhibited good tissue adhesion, rapid self-healing ability, and other multifunctions beneficial to wound healing, including killing of E. coli and S. aureus, rapid hemostasis and promoting migration of L929 cells. The full-thickness skin wound model shows that the hydrogel dressing significantly accelerated wound contraction, with increased granulation tissue thickness, collagen disposition, and enhanced vascularization, thus promoting wound healing. Therefore, the thiol-Michael addition reaction is an effective method for creating multifunctional hydrogels, and the injectable self-healing alginate/PEG hydrogels prepared in this way could be used in the biomedical area as wound healing dressing materials.

Impact nano- and micro- form of CdO on barley growth and oxidative stress response

The objective was investigated the effects of CdO and nano-CdO as potential toxic pollutants on growth and redox response of barley. CdO and nano-CdO have been found to cause significant phytotoxicity in barley seedlings, with nano-CdO increasing plant tissue cadmium accumulation. This accumulation is linked to growth retardation and oxidative stress. Low molecular weight antioxidants like restored glutathione and ascorbate have been found to increase the activity of glutathione peroxidase (GPX), glutathione-S-transferase (GSTs), and ascorbate peroxidase (APX) in green tissues. Catalase (CAT) activity increased from 50 % with 100 mg/l CdO to 70 % with 1000 mg/l and nano-CdO. The observed disturbance in redox balance signals the upregulation of corresponding genes. Antioxidant enzyme isoform gene transcripts increased for SODB, CAT2, and APX. Cadmium buildup in root cells causes oxidative stress, leading to upregulation of SOD, CAT, GR, and GSTs isoform genes as well as protein carbonylation, sulfhydryl group degradation, and MDA accumulation. CdO and nano-CdO have similar phytotoxic effects, but bioavailability affects biochemical and molecular responses.

Modification of pepper seed protein isolate to improve its functional characteristic by high hydrostatic pressure

Pepper seed protein isolate (PSPI) is a valuable plant-based protein source, yet the impact of processing methods such as high hydrostatic pressure (HHP) on its properties remains unclear. The impact of HHP on the structural and functional properties of PSPI at pH 7 and pH 9 was evaluated. Structural changes in PSPI were analyzed using spectral techniques, revealing significant alterations in the secondary and tertiary structures induced by HHP treatment. HHP treatment caused the unfolding of the PSPI structure, leading to the exposure of previously hidden chromophores and hydrophobic groups. The treatment also led to changes in free sulfhydryl groups and increased average particle size suggesting the formation of macromolecular polymers or insoluble aggregates. Consequently, the water-holding capacity, oil-holding capacity, foaming characteristics, and emulsifying activity index of the modified PSPI were significantly enhanced both at pH 7 and pH 9, with maximum improvements of 121.98 %, 157.29 %, 100.00 %, and 265.78 %, respectively. In conclusion, HHP is a promising strategy for enhancing the physicochemical properties of PSPI for various applications.

Studies on the complex interactions of different egg yolk proteins and their roles on rheological properties and stability of high internal phase Pickering emulsions

Egg yolks (EY) are widely used in food emulsions due to their high contents of amphiphilic proteins. This study investigated the different characterization and interactions among low-density lipoproteins (LDL), high-density lipoproteins (HDL) and phosvitins (Pv) using fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), scanning electron microscopy (SEM). The high internal-phase Pickering emulsions (HIPPEs) prepared by those protein complexes were analyzed on the basis of droplet size, optical microstructure, confocal microscopy images, interfacial protein compositions and rheological behavior. Results showed that LDL had a smaller particle size (56.44 ± 1.27 nm) and a greater ability to reduce interfacial tension, resulting in smaller droplet sizes (7.09 ± 0.25 μm) in LDL-stabilized HIPPEs. In contrast, HDL possessed higher hydrophobicity and a greater number of free sulfhydryl groups. The droplets in HDL-stabilized HIPPEs were larger, more compact, and exhibited lower solid lipid balance values in rheological tests. The competing adsorption of HDL and LDL at the interface influenced the properties of HDL-LDL prepared emulsions. However, an appropriate amount of Pv promoted the formation of an electrical double layer between LDL and HDL, further reducing interfacial tension. LDL-HDL-Pv complex stabilized HIPPEs displayed more gel-like rheological properties and the highest stability during the storage. This study provides insights for the development of EY protein-based emulsification systems in food industry.

Differentiated texture and lipid-lowering effect of cooked pork belly by different cooking regimes: insights from myofibrillar protein aggregation and dissociation, micromorphology

The effects of traditional flame stewing (TFS), commercial pressure cooking (PC), and sous-vide cooking (SVC) on the texture of pork belly and the underlying mechanism were investigated. SVC treatment at a low temperature (60 °C) achieved weaker oxidation of sulfhydryl groups, lower α-helical loss, higher surface hydrophobicity, and actomyosin dissociation, which reduced the shrinkage of myofibrils and loss of immobilized water, thus contributed to a tender meat. However, the protein aggregation predominated without actomyosin dissociation during PC treatment, causing the toughening of meat. TFS and PC intensified the atrophy of adipocytes and damage of cell membranes, which helped for the softening of adipose tissue and achieved 13.82% and 17.62 % lipid loss, respectively, and contributed to a higher ratio of polyunsaturated fatty acids to saturated fatty acids (0.45–0.46) than SVC (0.34). Comparisons showed that TFS was a more suitable method to prepare cooked pork belly with tender and juicy texture, low lipid content, and high ratio of polyunsaturated fatty acids to saturated fatty acids. This study offered new insights into how cooking regimes affect the texture and lipid-lowering effects of stewed pork.

Heterologous expression and purification of glutamate decarboxylase-1 from the model plant Arabidopsis thaliana: Characterization of the enzyme's in vitro truncation by thiol endopeptidase activity

Plant glutamate decarboxylase (GAD) is a Ca2+-calmodulin (CaM) activated enzyme that produces γ-aminobutyrate (GABA) as the first committed step of the GABA shunt. Our prior research established that in vivo phosphorylation of AtGAD1 (AT5G17330) occurs at multiple N-terminal serine residues following Pi resupply to Pi-starved cell cultures of the model plant Arabidopsis thaliana. The aim of the current investigation was to purify recombinant AtGAD1 (rAtGAD1) following its expression in Escherichia coli to facilitate studies of the impact of phosphorylation on its kinetic properties. However, in vitro proteolytic truncation of an approximate 5 kDa polypeptide from the C-terminus of 59 kDa rAtGAD1 subunits occurred during purification. Immunoblotting demonstrated that most protease inhibitors or cocktails that we tested were ineffective in suppressing this partial rAtGAD1 proteolysis. Although the thiol modifiers N-ethylmaleimide or 2,2-dipyridyl disulfide negated rAtGAD1 proteolysis, they also abolished its GAD activity. This indicates that an essential -SH group is needed for catalysis, and that rAtGAD1 is susceptible to partial degradation either by an E. coli cysteine endopeptidase, or possibly via autoproteolytic activity. The inclusion of exogenous Ca2+/CaM facilitated the purification of non-proteolyzed rAtGAD1 to a specific activity of 27 (μmol GABA produced/mg) at optimal pH 5.8, while exhibiting an approximate 3-fold activation by Ca2+/CaM at pH 7.3. By contrast, the purified partially proteolyzed rAtGAD1 was >40 % less active at both pH values, and only activated 2-fold by Ca2+/CaM at pH 7.3. These results emphasize the need to diagnose and prevent partial proteolysis before conducting kinetic studies of purified regulatory enzymes.

Effect of chitosan on thermal gelling properties of pork myofibrillar protein and its mechanism.

Previous studies have demonstrated that the addition of chitosan can improve the quality and functional properties of meat products. However, the underlying mechanism remains unclear. In this study, the effect and mechanism of the addition of chitosan on the gel properties of myofibrillar protein (MP) were investigated. The results indicated that the gel strength and the water-holding capacity of MP-chitosan gel increased significantly when chitosan was added at 2.5-10 mg mL-1. Myofibrillar protein samples with 10 mg mL-1 added chitosan exhibited the highest elasticity and viscosity during gel formation and strengthening. The addition of chitosan also caused a modification in both the secondary and tertiary structure of MP, resulting in an enhanced exposure of hydrophobic and sulfhydryl groups in comparison with the control. Chitosan inhibited the conversion of immobilized water into free water and the formation of water channels during the thermal gelation process of MP. The denaturation enthalpy (ΔH) of myosin decreased as the concentration of chitosan exceeded 5 mg mL-1. The microstructure showed that the incorporation of chitosan (5-10 mg mL-1) facilitated the formation of compact and well organized MP gel networks. The addition of chitosan can enhance the functional properties of meat protein and facilitate heat-induced gelation, making it a promising ingredient for improving the quality of processed meat products. © 2024 Society of Chemical Industry.