Dodecyl Sulfate-polyacrylamide Gel Electrophoresis Research Articles

Regulation of Amylose Content by Single Mutations at an Active Site in the Wx-B1 Gene in a Tetraploid Wheat Mutant.

The granule-bound starch synthase I (GBSSI) encoded by the waxy gene is responsible for amylose synthesis in the endosperm of wheat grains. In the present study, a novel Wx-B1 null mutant line, M3-415, was identified from an ethyl methanesulfonate-mutagenized population of Chinese tetraploid wheat landrace Jianyangailanmai (LM47). The gene sequence indicated that the mutated Wx-B1 encoded a complete protein; this protein was incompatible with the protein profile obtained using sodium dodecyl sulfate–polyacrylamide gel electrophoresis, which showed the lack of Wx-B1 protein in the mutant line. The prediction of the protein structure showed an amino acid substitution (G470D) at the edge of the ADPG binding pocket, which might affect the binding of Wx-B1 to starch granules. Site-directed mutagenesis was further performed to artificially change the amino acid at the sequence position 469 from alanine (A) to threonine (T) (A469T) downstream of the mutated site in M3-415. Our results indicated that a single amino acid mutation in Wx-B1 reduces its activity by impairing its starch-binding capacity. The present study is the first to report the novel mechanism underlying Wx-1 deletion in wheat; moreover, it provided new insights into the inactivation of the waxy gene and revealed that fine regulation of wheat amylose content is possible by modifying the GBSSI activity.

Gum tragacanth-sodium alginate active coatings containing epigallocatechin gallate reduce hydrogen peroxide content and inhibit lipid and protein oxidations of large yellow croaker (Larimichthys crocea) during superchilling storage

The active coatings supplemented with epigallocatechin gallate (EGCG) (0.16 %, 0.32 %, and 0.64 %, respectively) combined with superchilling storage (−3 ± 0.2 °C) were used to reduce hydrogen peroxide (H2O2) content, and inhibit lipid and protein oxidations of large yellow croaker during 42 days of superchilling storage. EGCG coatings delayed lipid and protein oxidations by inhibiting the generation of H2O2, malondialdehyde (MDA) and carbonyl groups, and maintaining a higher Ca2+-ATPase activity and sulfhydryl content. We also observed that EGCG treatments maintained myofibrillar organized secondary structure by keeping higher α-helix content, and also stabilized tertiary structure during superchilling storage. Low-field nuclear magnetic resonance (LF-NMR) revealed that EGCG treatments might improve the association of water molecules with protein for fixed water. In addition, sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and scanning electron microscope (SEM) images both showed that these treatments could delay the myofibrillar degradation of fresh fish. Overall, we report that the active coatings containing EGCG treatments protect the lipid and protein of large yellow croaker during superchilling storage.

Effect of pH on the conformational structure of cytochrome c and subsequent enzymatic cross-linking catalyzed by laccase

BackgroundThe aim of the present study was to investigate the effect of substrate conformational structure changes on the laccase-induced protein cross-linking. The effects of laccase amount, pH, and ferulic acid (FA) on the enzymatic cross-linking of substrate, Cyt C, were determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. High-performance size exclusion chromatography, laser particle size analysis and isothermal titration calorimetry (ITC) were also applied to investigate the cross-linking product and enthalpy changes. Structural changes of Cyt C at different pH values were analyzed by ultraviolet–visible (UV–vis), fluorescence, and circular dichroism (CD) measurements. ResultsComplete cross-linking, partial cross-linking, minute cross-linking, and no cross-linking occurred at pH 2.0, 4.0, 6.0, and 8.0, respectively. ITC analysis demonstrated that the enzymatic cross-linking of Cyt C was an endothermic process. The UV–vis, fluorescence, and CD measurements exhibited that the tertiary structure of Cyt C was disrupted, and part of the α-helical polypeptide region unfolded at pH 2.0. The structural flexibilities decreased, and the tertiary structure of Cyt C became increasingly compact with the increase in pH values from 4.0 to 8.0. The gradual changes in the structure of Cyt C at different pH values were in accordance with the cross-linking results of Cyt C catalyzed by laccase. ConclusionsThe results demonstrated that minute structure changes of substrate had a remarkable effect on the laccase-induced cross-linking. The findings promote the understanding of the substrate requirement of laccase in protein cross-linking and are instructive for the modulation of laccase-induced protein cross-linking.How to cite: Li D-X, Qi Z-Y, Liu J-Y, et al. Effect of pH on the conformational structure of cytochrome c and subsequent enzymatic cross-linking catalyzed by laccase. Electron J Biotechnol 2022;60. https://doi.org/10.1016/j.ejbt.2022.07.002.

Light and prey influence the abundances of two rhodopsins in the dinoflagellate Oxyrrhis marina

Antisera were raised against the C-terminal amino acid sequences of the two rhodopsins ADY17806 and AEA49880 of Oxyrrhis marina. The antisera and affinity-purified antibodies thereof were used in western immunoblotting experiments of total cell protein fractions from cultures grown either in darkness or in white, red, green, or blue light. Furthermore, the rhodopsin abundances were profiled in cultures fed with yeast or the prasinophyte Pyramimonas grossii. The immunosignals of ADY17806 and AEA49880 were similar when O. marina was grown in white, green, or blue light. Signal intensities were lower under conditions of red light and lowest in darkness. Higher amounts were registered for both rhodopsins when O. marina was fed with yeast compared to P. grossii. Furthermore, total cell protein of cultures of O. marina grown under all cultivation conditions was separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, followed by tryptic in-gel digestion and mass spectrometric analysis of the 25-kDa protein bands. The rhodopsin ADY17809 was detected in all samples of the light quality experiments and in 14 of the 16 samples of the prey quality experiments. The rhodopsin ABV22427 was not detected in one sample of the light quality experiments. It was detected in 15 of the 16 samples of the prey quality experiments. Peptide fragments of the other rhodopsins were detected less often, and no clear distribution pattern was evident with respect to the applied light quality or offered prey, indicating that none of them was exclusively formed under a distinct light regime or when feeding on yeast or the prasinophyte. Fluorescence light microscopy using the affinity-purified antibodies revealed significant labeling of the cell periphery and cell internal structures, which resembled vacuoles, tiny vesicles, and rather compact structures. Immunolabeling electron microscopy strengthened these results and showed that the cytoplasmic membrane, putative lysosome membranes, membranes encircling the food vacuole, and birefringent bodies became labeled.

Bioactivation of 3D Cell-Imprinted Polydimethylsiloxane Surfaces by Bone Protein Nanocoating for Bone Tissue Engineering.

Physical and chemical parameters that mimic the physiological niche of the human body have an influence on stem cell fate by creating directional signals to cells. Micro/nano cell-patterned polydimethylsiloxane (PDMS) substrates, due to their ability to mimic the physiological niche, have been widely used in surface modification. Integration of other factors such as the biochemical coating on the surface can achieve more similar microenvironmental conditions and promote stem cell differentiation to the target cell line. Herein, we investigated the effect of physical topography, chemical functionalization by acid bone lysate (ABL) nanocoating, and the combined functionalization of the bone proteins’ nanocoated surface and the topographically modified surface. We prepared four distinguishing surfaces: plain PDMS, physically modified PDMS by 3D cell topography patterning, chemically modified PDMS with bone protein nanocoating, and chemically modified nano 3D cell-imprinted PDMS by bone proteins (ABL). Characterization of extracted ABL was carried out by Bradford staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis, followed by the MTT assay for evaluation of cell viability on ABL-coated PDMS. Moreover, field emission scanning electron microscopy and profilometry were used for the determination of optimal coating thickness, and the appropriate coating concentration was identified and used in the study. The binding and retention of ABL to PDMS were confirmed by Fourier transform infrared spectroscopy and bicinchoninic acid assay. Sessile drop static water contact angle measurements on substrates showed that the combined chemical functionalization and nano 3D cell-imprinting on the PDMS surface improved surface wettability by 66% compared to plain PDMS. The results of ALP measurement, alizarin red S staining, immunofluorescence staining, and real-time PCR showed that the nano 3D cell-imprinted PDMS surface functionalized by extracted bone proteins, ABL, is able to guide the fate of adipose derived stem cellss toward osteogenic differentiation. Eventually, chemical modification of the cell-imprinted PDMS substrate by bone protein extraction not only improved the cell adhesion and proliferation but also contributed to the topographical effect itself and caused a significant synergistic influence on the process of osteogenic differentiation.

Characterization of whey protein isolate-(−)-epigallocatechin-3-gallate conjugates prepared by non-enzymatic and enzymatic methods and their application in stabilizing β-carotene emulsion

(−)-Epigallocatechin-3-gallate (EGCG) was conjugated to whey protein isolate (WPI) using alkaline treatment, free radical grafting and tyrosinase-catalyzed oxidation. The structure and properties of the obtained conjugates and their application as antioxidant emulsifiers in stabilizing β-carotene emulsion were systematically characterized. Fourier transform infrared spectroscopy and sodium dodecyl sulfate–polyacrylamide gel electrophoresis verified the covalent linking between WPI and EGCG. The highest grafting efficiency was obtained for enzyme conjugate, followed for alkaline conjugate. Conjugation of EGCG decreased the α-helix content and fluorescence intensity of protein, and these changes depend on both EGCG conjugation and the cross-linking methods. Due to their improved emulsifying properties and antioxidant activity, WPI-EGCG conjugates formed smaller emulsion droplets and protect the encapsulated β-carotene more effectively, and the protective property is positively correlated with EGCG content in conjugates.

Magnetic Levitation Patterns of Microfluidic-Generated Nanoparticle-Protein Complexes.

Magnetic levitation (MagLev) has recently emerged as a powerful method to develop diagnostic technologies based on the exploitation of the nanoparticle (NP)–protein corona. However, experimental procedures improving the robustness, reproducibility, and accuracy of this technology are largely unexplored. To contribute to filling this gap, here, we investigated the effect of total flow rate (TFR) and flow rate ratio (FRR) on the MagLev patterns of microfluidic-generated graphene oxide (GO)–protein complexes using bulk mixing of GO and human plasma (HP) as a reference. Levitating and precipitating fractions of GO-HP samples were characterized in terms of atomic force microscopy (AFM), bicinchoninic acid assay (BCA), and one-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis (1D SDS-PAGE), and nanoliquid chromatography–tandem mass spectrometry (nano-LC-MS/MS). We identified combinations of TFR and FRR (e.g., TFR = 35 μL/min and FRR (GO:HP) = 9:1 or TFR = 3.5 μL/min and FRR (GO:HP) = 19:1), leading to MagLev patterns dominated by levitating and precipitating fractions with bulk-like features. Since a typical MagLev experiment for disease detection is based on a sequence of optimization, exploration, and validation steps, this implies that the optimization (e.g., searching for optimal NP:HP ratios) and exploration (e.g., searching for MagLev signatures) steps can be performed using samples generated by bulk mixing. When these steps are completed, the validation step, which involves using human specimens that are often available in limited amounts, can be made by highly reproducible microfluidic mixing without any ex novo optimization process. The relevance of developing diagnostic technologies based on MagLev of coronated nanomaterials is also discussed.

Impact of Escherichia coli Outer Membrane Vesicles on Sperm Function.

Reproductive tract infections account for approximately 15% of male infertility cases. Escherichia coli (E. coli) represents the most frequently isolated bacterial strain in the semen of infertile men. All Gram-negative bacteria constitutively produce outer membrane vesicles (OMVs). The present study proved, for the first time, the involvement of OMVs in human sperm function. E. coli OMVs were isolated by ultracentrifugation and characterized via sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis. Human sperm was exposed to OMVs (8 µg/mL) for different times (30, 45, 60 and 90 min). The vitality, motility, morphology, ROS level and DNA fragmentation of spermatozoa were evaluated. OMVs reduced the progressive motility and increased the immobile spermatozoa amount after 30 min of treatment. In addition, a significant increase in the percentage of intracellular ROS and sperm DNA fragmentation was recorded for each vesicular exposure time. These preliminary findings prove that OMVs contribute to altering human sperm function via two mechanisms: (i) impaired motility and (ii) DNA fragmentation.

Application of Aqueous Two-Phase System to the Purification of Persimmon Polyphenol Oxidase

Polyphenol oxidases (PPOs), which have recently become very popular among many researchers, catalyze the oxidation of phenolic compounds. Polyphenol oxidases are generally found in plants. Among them, persimmon (Diospyros kaki L.) is known as a good source for polyphenol oxidases. In this study, the polyphenol oxidase enzyme was purified from persimmon fruit using aqueous two-phase (ATPS). The optimized system was composed of 18% (w/w) PEG4000, 7% (w/w) NaH2 P04 and 1% (w/w) NaCl (pH 8.5, 25°C and 5 g). The PPO enzyme was obtained from the system by 4.8-fold purification with 191% activity recovery. The molecular weight of the enzyme, 28 kDa, was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. In summary, it has been shown that the PPO enzyme can be obtained from persimmon in a short time and at low cost using a non-chromatographic method-ATPS.

Highly selective titanium (IV)-immobilized O-phospho-L-tyrosine modified magnetic nanoparticles for the enrichment of intact phosphoproteins.

Phosphorylation is one of the most important protein post-translational modifications, which possesses dramatic regulatory effects on the function of proteins. In consideration of the low abundance and low stoichiometry of phosphorylation and non-specific signal suppression, efficient capture of the phosphoproteins from complex biological samples is critical to meet the need for protein profiling. In this work, a facile preparation of titanium (IV)-immobilized O-phospho-L-tyrosine modified magnetic nanoparticles was developed for the enrichment of intact phosphoproteins. The prepared magnetic nanoparticles were characterized by various instruments and had a spherical shape with an average diameter of 300 nm. The adsorption isotherms were investigated and the maximum capacity for β-casein was calculated to be 961.5 mg/g. Standard protein mixtures and biological samples (non-fat milk and human serum) were selected to test the enrichment performance. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis demonstrated the excellent enrichment performance with high selectivity. With the superparamagnetic property, titanium (IV)-immobilized O-phospho-L-tyrosine modified magnetic nanoparticles were convenient for the practical application and clinical promotion, thus having a promising prospect in the field of phosphoprotein research.

The effects of tea polyphenol-ozonated slurry ice treatment on the quality of large yellow croaker (Pseudosciaena crocea) during chilled storage.

The aim of the current study was to evaluate the synergistic effects of tea polyphenol-ozonated slurry ice on the quality, physicochemical and protein characteristics of large yellow croaker (Pseudosciaena crocea) during chilled (4°C) storage. To 0.3% tea polyphenol combined with ozone water was added sodium chloride until the salt concentration reached 3.3% and with the use of an ice machine the mixture formed the tea polyphenol-ozonated slurry ice. Microbial [total viable count (TVC)], physicochemical [total volatile basic nitrogen (TVB-N), K value], myofibrillar fragmentation index (MFI), Ca2+ -ATPase activity, total sulfhydryl content, intrinsic fluorescence intensity (IFI), Fourier-transform infrared (FTIR), scanning electron microscopy (SEM) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) were analyzed during chilled (4°C) storage for up to 20 days. The results showed that tea polyphenol-ozonated slurry ice could effectively inhibit the increase of TVC and TVB-N, reduce the degree of adenosine triphosphate (ATP) degradation. In addition, the tea polyphenol-ozonated slurry ice treatment could protect the integrity of myosin in myofibrillar proteins (MPs) by inhibiting the decrease of Ca2+ -ATPase activity and the content of total sulfhydryl. Furthermore, the tea polyphenol-ozonated slurry ice presented a superiorly protective effect on protein structure in MPs as manifested by the results of IFI, FTIR and SDS-PAGE. It was possible that due to the addition of tea polyphenol, the antioxidant activity of this complex was significantly improved. The tea polyphenol-ozonated slurry ice treatment can maintain the quality of large yellow croaker by decreasing the damage of MP caused by the interaction between microorganisms and endogenous enzymes. © 2022 Society of Chemical Industry.

Disinfection effect and mechanism of neutral electrolyzed water on Staphylococcus albus

To evaluate the disinfection efficacy and mechanism of neutral electrolyzed water(NEW) on Staphylococcus albus. The bactericidal effect of NEW was observed through suspension quantitative bactericidal test. In addition, to analyze the mechanism of NEW, the leakage of bacterial contents and enzyme activity were tested, and the damage of bacterial protein and DNA were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and agarose gel electrophoresis. After treated with NEW for 1 min, staphylococcus albus in suspension(2.07×10~7 CFU/mL)were reduced to below the detection limit(<10 CFU/mL), with killing rate >99.999% and killing log value >5. NEW could destroy the integrity of the cell membrane, resulting in the leakage of Ca~(2+), K~+, protein and nucleic acid, and change the activity of superoxide dismutase(SOD), peroxidase(POD), catalase(CAT) and other enzymes, it could also damage protein and DNA. NEW can effectively kill Staphylococcus albus in suspension, and its mechanism involves the destruction of cell membrane, bacterial protein and nucleic acid, and the change of enzyme activity.

Molecular cloning, characterization, and homology modeling of serine hydroxymethyltransferase from psychrophilic bacterium Psychrobacter sp.

Serine hydroxymethyltransferase (SHMT) plays a significant role in the synthesis of l-serine, purine, and thymidylate, which could be extensively applied in the treatment of cancers and the development of antibiotics. In this study, cloned from Psychrobacter sp. ANT206, a novel cold-adapted SHMT gene (psshmt, 1257 bp) encoding a protein of 418 amino acids was expressed in Escherichia coli. The homology modeling result revealed that PsSHMT owned fewer Proline (Pro) residues and hydrogen bonds compared with its homologs from mesophilic E. coli and thermophilic Geobacillus stearothermophilus. In addition, the molecular weight of the purified recombinant PsSHMT (rPsSHMT) was identified to be 45 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis, approximately. The enzymatic characteristics of the cold-adapted rPsSHMT displayed that its optimum temperature and pH were 30°C and 7.5, respectively, and its enzymatic activity could be inhibited by Cu2+ , significantly. rPsSHMT also showed a high kcat value and low ΔG at low temperatures. Furthermore, arginine (Arg) could affect the activity of rPsSHMT and be vital to its active sites. The results of this study reflected that these characteristics of the cold-adapted rPsSHMT made it a remarkable candidate that could be utilized in multiple industrial fields under low temperatures.

Detection of Echinococcus granulosus sensu lato cysts and seroprevalence of cystic echinococcosis in cattle and camels in Maiduguri Abattoir.

Cystic echinococcosis (CE) is a zoonotic disease of great importance worldwide. This study was conducted to determine the prevalence and antigenic profile of Echinococcus cysts (CE cysts) in camels and cattle. The lungs, livers, hearts, and kidneys of 560 animals, comprising 304 camels and 256 cattle slaughtered in the Maiduguri abattoir, were examined for CE. Blood samples were collected for serology. Protein profiles of CE fluids were analyzed using indirect Enzyme Linked Immunosorbent Assay while Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) was used to characterize the electrophoretic pattern of different CE cyst fluid samples. The overall prevalence of CE was 8.4%, 14.14% (95% CI = 10.65-18.54%) in camels and 1.60% (95% CI = 0.46-4.09%) in cattle. Adult camels 41 (16.21%) (95% CI = 12.15-21.27%) had a higher prevalence than the young camels 2 (3.92%) (95% CI = 0.33-13.97%) (p = 0.038). In cattle, only adults 4 (2.0%) had cysts. Higher prevalence of CE was recorded in male 22 (16.42%) (95% CI = 11.03-23.68%) than female 21 (12.35%) (95% CI = 8.16-18.21%) camels [p = 0.399] while only female cattle 3 (2.2%) had cysts. Higher prevalence of CE was recorded in the livers of 34 (11.18%) (95% CI = 8.08-15.25%) than in the lungs 25 (8.22%) (95% CI = 5.59-11.90%) of camels [p = 0.273]. Of the 47 cysts collected, 43 (91.49%) and 4 (8.51%) were from camels and cattle, respectively. A total of 18 (38.30%) fertile, 17 (36.17%) non-fertile, and 12 (25.53%) calcified cysts were recovered in animals. Overall seroprevalence of 52.63% (95% CI = 47.02-58.18%) and 35.55% (95% CI = 29.93-41.59%) were observed in camels and cattle in this study. The SDS-PAGE of camel CE cyst fluids revealed protein bands at 64kda, 91kda, 160kda, and 200kda molecular units while the purified cyst fluids revealed bands at 64kda, 91kda, 120kda, 160kda, and 200kda. Regular meat inspections and the exclusion of dogs from abattoir premises are strongly encouraged. Investigation into local prevailing factors encouraging transmission should be carried out.

Gene Expression and Characterization of Iturin A Lipopeptide Biosurfactant from Bacillus aryabhattai for Enhanced Oil Recovery.

Biosurfactants are eco-friendly surface-active molecules recommended for enhanced oil recovery techniques. In the present study, a potential lipopeptide (biosurfactant) encoding the iturin A gene was synthesized from Bacillus aryabhattai. To improvise the yield of the lipopeptide for specific applications, current research tends toward engineering and expressing recombinant peptides. An iturin A gene sequence was codon-optimized, amplified with gene-specific primers, and ligated into the pET-32A expression vector to achieve high-level protein expression. The plasmid construct was transformed into an E. coli BL21 DE3 host to evaluate the expression. The highly expressed recombinant iturin A lipopeptide was purified on a nickel nitrilotriacetic acid (Ni-NTA) agarose column. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the purity and molecular mass of iturin A was 41 kDa. The yield of recombinant iturin A was found to be 60 g/L with a 6.7-fold increase in comparison with our previously published study on the wild strain. The approach of cloning a functional fragment of partial iturin A resulted in the increased production of the lipopeptide. When motor oil was used, recombinant protein iturin A revealed a biosurfactant property with a 74 ± 1.9% emulsification index (E24). Purified recombinant protein iturin A was characterized by mass spectrometry. MALDI-TOF spectra of trypsin digestion (protein/trypsin of 50:1 and 25:1) showed desired digested mass peaks for the protein, further confirming the identity of iturin A. The iturin A structure was elucidated based on distinctive spectral bands in Raman spectra, which revealed the presence of a peptide backbone and lipid. Recombinant iturin A was employed for enhanced oil recovery through a sand-packed column that yielded 61.18 ± 0.85% additional oil. Hence, the novel approach of the high-level expression of iturin A (lipopeptide) as a promising biosurfactant employed for oil recovery from Bacillus aryabhattai is not much reported. Thus, recombinant iturin A demonstrated its promising ability for efficient oil recovery, finding specific applications in petroleum industries.

Purification and partial characterization of a novel lectin from Ruta montana roots

Lectins are proteins or glycoproteins, which participate in various biological processes. The purpose of this study was to purify new lectin from Ruta montana roots. Lectin of Ruta montana has been purified from roots using ultrafiltration and precipitation with ammonium sulfate followed by gel filtration chromatography whereas protein estimation was done by Bradford’s method. Within sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE) analysis, the extract exhibited three bands and one band after purification. The molecular weight of lectin was determined by SDS–PAGE and gel filtration chromatography, which was found to be a monomeric protein of approximately 28.8 kDa. The agglutination activity of Ruta montana lectins was stable within a temperature range from 4 to 50° C for 30 min and the pH range from 4.2 to 9. This study presents a natural source of lectins that can be used in several other studies due to its different biological activities.

Effect of ball milling-assisted glycosylation modification on the structure and foaming property of egg white protein.

The effect of different glycosylation degrees on molecular structure and foaming property of egg white protein (EWP) was investigated using ball milling-assisted glycosylation. The results showed the foaming ability (FA) and foam stability (FS) of EWP improved when the degree of glycosylation was increased. In particular, FA of ball milling-assisted glycosylation of EWP enhanced by 39.9% and 28.8%, and the FS increased by 28.7% and 24.0% compared with EWP and ball milling egg white protein (BE) at 150min of reaction. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis could reflect the grafting degree of EWP and glucose molecules from the side. When EWP was fully grafted with glucose, endogenous fluorescence and free sulfhydryl groups indicated that tertiary structure of EWP was depolymerized, and Fourier transform infrared spectroscopy showed the secondary structure tended to change from order to disorder. The results of this study indicated that ball milling-assisted glycosylation modification was a practical method to improve the foaming property of EWP. PRACTICAL APPLICATION: EWP has great FA and FS, making it indispensable in the baking industry. In this study, ball milling-assisted glycosylation was used to improve the foaming property of EWP, and the molecular structure of EWP with different degrees of glycosylation was fully resolved. The results demonstrated that ball milling, as a physical pretreatment, can fully unfold the structure of EWP. When sugar molecules were fully grafted, the particle size of EWP reduced, solubility increased, and the stability of system improved, thus enhancing the foaming property of EWP. The results can provide theoretical basis for improving the foaming property of EWP and provide a reference value for its industrial application.

Effect of Storage Conditions on the Protein Composition and Structure of Peanuts.

Peanuts are important oil crops and plant protein source. This study evaluated the influence of storage temperature (15, 25, and 35 °C) and time (0, 160, and 320 days) on the protein composition and the molecular structure of peanuts through sodium dodecyl sulfate polyacrylamide gel electrophoresis, particle size, total sulfhydryl (−SH) contents, nanostructural characterization, surface morphology microstructure, and spatial distribution of proteins and lipid analysis. Results showed that the basic subunits and disulfide contents of peanut protein were not affected by storage temperature and time. However, the −SH contents decreased significantly (P < 0.05) in all samples except the 15 °C/160 day storage group. The protein particle size and graininess increased when stored at 25 and 35 °C for 160 and 320 days, respectively; however, there was no significant change (P > 0.05) when stored at 15 °C. In addition, significant changes (P < 0.05) on the microscopic morphology and spatial distribution of protein and lipids were observed when stored at 25 and 35 °C for 320 days.

Nanoparticle formulation of the fusion protein virus like particles of respiratory syncytial virus stimulates enhanced in vitro antigen presentation and autophagy

Respiratory Syncytial Virus (RSV) is one of the leading causes of bronchiolitis and pneumonia in childrenunder one year globally. As a result, RSV poses a severe burden on healthcare services. Thus, a vaccine for RSV is a global need. Utilizing polymeric nanoparticles as a delivery system for vaccine antigen holds a lot of promise. In this study, the virus like particles of RSV fusion protein (F-VLP) was encapsulated in poly (D, L-lactide-co-glycolide) (PLGA) nanoparticles (NP). The F-VLP NP was formulated using a double emulsion solvent evaporation technique. The optimized NPs had a particle size of 525 ± 10.5 nm and an antigen encapsulation efficiency of 73% ± 10.5. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis showed that the F-VLP was stable post formulation. The F-VLP NP showed a sustained release of the F-VLP antigen for up to a week. In vitro study revealed that the F-VLP NP were non-cytotoxic, and the cellular uptake of the NPs by dendritic cells was observed within 3 h. The F-VLP NP with adjuvant monophosphoryl lipid A (MPL) NP and without MPL NP showed enhanced expression of antigen presentation molecule major histocompatibility complex (MHC)-I and autophagosomes in dendritic cells. In summary, the sustained release of the antigen from the F-VLP NP and the particulate nature of the vaccine resulted in enhanced antigen presentation and induction of autophagy in antigen-presenting cells (APCs).

Gelatin From Cartilage of Siberian Sturgeon (Acipenser baerii): Preparation, Characterization, and Protective Function on Ultraviolet-A-Injured Human Skin Fibroblasts

For full use of the by-products during Siberian sturgeon (Acipenser baerii) processing, gelatin was extracted from the cartilages using the hot water method, and its physico-chemical properties and protective function on ultraviolet-A injured human skin fibroblasts (HFSBs) were measured. Using single-factor and orthogonal experiments, the conditions for extracting gelatin from Siberian sturgeon cartilage were optimized as extraction time of 7 h, pH 9, material-to-liquid ratio (g/ml) of 1:5, and temperature of 45°C. The prepared gelatin (TCG) with a yield of 28.8 ± 1.53% had Gly (307 residues/1,000 residues) as the major amino acid and contained a lower amount (214 residues/1,000 residues) of imino acids than that (227 residues/1,000 residues) of pigskin gelatin (PSG). Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), ultraviolet spectrum, and infrared spectroscopy analysis indicated that TCG had the main spectral characteristics of fish gelatin and contained α chains (α1 and α2 chains) and β chain of type I collagen, but its structural stability was lower than that of PSG due to its low content of imino acids, which induced the smaller molecular bands observed in the SDS-PAGE pattern. TCG exhibited lower water content, gel strength, emulsion stability index, foam capacity, foam stability, and water-holding capacity but higher ash content, transmittance, emulsion activity index, and fat-binding capacity (P &amp;lt; 0.05). Moreover, TCG could significantly protect HFSBs against ultraviolet-A injury by enhancing the activity of superoxide dismutase, catalase, and glutathione peroxidase to scavenge excess reactive oxygen species and decrease the content of malondialdehyde. Therefore, gelatin from Siberian sturgeon cartilages could act as promising candidates when applied in health-promoting products against ultraviolet-A injury.