Changes In Protein Composition Research Articles

Steam cooking drives alterations of proteomics, protein structural and functional properties in rice (Oryza sativa l.)

Rice, as an important staple food consumed by people, is gradually becoming a concern for many consumers in terms of its eating quality. Many studies have found that protein is an important factor to determine the edible quality of rice. However, the protein changes during rice cooking have not been compared and studied in detail. In this study, the structural and functional characteristics of rice proteins before and after steaming were comprehensively compared. Meanwhile, proteomic strategies were also applied to analyze changes in protein composition and expression. Steaming promoted the aggregation of cooked rice proteins, resulting in a more compact structure and a significant increase in particle size (raw rice, 304.33 nm; cooked rice, 565.17 nm). Compared with raw rice protein (43.33%; 133.91%), the solubility (22.29%) and foaming capacity (119.53%) of cooked rice protein were significantly reduced, while emulsion activity was slightly improved. The differential abundance proteins were found to be predominantly down-regulated after steaming (238 proteins) and mostly enriched in metabolic pathways. The steaming process of rice mainly involves oxidative phosphorylation, thiamine metabolism, ether lipid metabolism, glycerophospholipid metabolism, multiple amino acid metabolism and other metabolic pathways, and they are closely related to each other. This study deepens the understanding of proteomic changes and structural/functional properties of rice proteins during steaming.

Mitochondrial Ribosomal Protein MRPS15 Is a Component of Cytosolic Ribosomes and Regulates Translation in Stressed Cardiomyocytes.

Regulation of mRNA translation is a crucial step in controlling gene expression in stressed cells, impacting many pathologies, including heart ischemia. In recent years, ribosome heterogeneity has emerged as a key control mechanism driving the translation of subsets of mRNAs. In this study, we investigated variations in ribosome composition in human cardiomyocytes subjected to endoplasmic reticulum stress induced by tunicamycin treatment. Our findings demonstrate that this stress inhibits global translation in cardiomyocytes while activating internal ribosome entry site (IRES)-dependent translation. Analysis of translating ribosome composition in stressed and unstressed cardiomyocytes was conducted using mass spectrometry. We observed no significant changes in ribosomal protein composition, but several mitochondrial ribosomal proteins (MRPs) were identified in cytosolic polysomes, showing drastic variations between stressed and unstressed cells. The most notable increase in polysomes of stressed cells was observed in MRPS15. Its interaction with ribosomal proteins was confirmed by proximity ligation assay (PLA) and immunoprecipitation, suggesting its intrinsic role as a ribosomal component during stress. Knock-down or overexpression experiments of MRPS15 revealed its role as an activator of IRES-dependent translation. Furthermore, polysome profiling after immunoprecipitation with anti-MRPS15 antibody revealed that the "MRPS15 ribosome" is specialized in translating mRNAs involved in the unfolded protein response.

Comparison of miR-106b, miR-191, and miR-30d expression dynamics in milk with regard to its composition in Holstein and Ayrshire cows.

Milk composition varies considerably and depends on paratypical, genetic, and epigenetic factors. MiRNAs belong to the class of small non-coding RNAs; they are one of the key tools of epigenetic control because of their ability to regulate gene expression at the post-transcriptional level. We compared the relative expression levels of miR-106b, miR-191, and miR-30d in milk to demonstrate the relationship between the content of these miRNAs with protein and fat components of milk in Holstein and Ayrshire cattle. Milk fat, protein, and casein contents were determined in the obtained samples, as well as the content of the main fatty acids (g/100 g milk), including: saturated acids, such as myristic (C14:0), palmitic (C16:0), and stearic (C18:0) acids; monounsaturated acids, including oleic (C18:1) acid; as well as long-, medium- and short-chain, polyunsaturated, and trans fatty acids. Real-time stem-loop one-tube reverse transcription polymerase chain reaction with TaqMan probes was used to measure the miRNA expression levels. The miRNA expression levels in milk samples were found to be decreased in the first two months in Holstein breed, and in the first four months in Ayrshire breed. Correlation analysis did not reveal any dependence between changes in the expression level of miRNA and milk fat content, but showed a multidirectional relationship with individual milk fatty acids. Positive associations between the expression levels of miR-106b and miR-30d and protein and casein content were found in the Ayrshire breed. Receiver operating characteristic curve analysis showed that miR-106b and miR-30d expression levels can cause changes in fatty acid and protein composition of milk in Ayrshire cows, whereas miR-106b expression level determines the fatty acid composition in Holsteins. The data obtained in this study showed that miR-106b, miR-191, and miR-30d expression levels in milk samples have peculiarities associated with breed affiliation and the lactation period.

The Interplay of Protein Hydrolysis and Ammonia in the Stability of Hevea Rubber Latex during Storage.

Natural rubber (NR) latex derived from Hevea brasiliensis is a complex colloid comprising mainly rubber hydrocarbons (latex particles) and a multitude of minor non-rubber constituents such as non-rubber particles, proteins, lipids, carbohydrates, and soluble organic and inorganic substances. NR latex is susceptible to enzymatic attack after it leaves the trees. It is usually preserved with ammonia and, to a lesser extent, with other preservatives to enhance its colloidal stability during storage. Despite numerous studies in the literature on the influence of rubber proteins on NR latex stability, issues regarding the effect of protein hydrolysis in the presence of ammonia on latex stability during storage are still far from resolved. The present work aims to elucidate the interplay between protein hydrolysis and ammoniation in NR latex stability. Both high- and low-ammonia (with a secondary preservative) NR latexes were used to monitor the changes in their protein compositions during storage. High-ammonia (FNR-A) latex preserved with 0.6% (v/v) ammonia, a low 0.1% ammonia/TMTD/ZnO (FNR-TZ) latex, and a deproteinized NR (PDNR) latex were labeled with fluorescence agents and observed using confocal laser scanning microscopy to determine their protein composition. Protein hydrolysis was confirmed via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The results revealed that protein hydrolysis increased with the storage duration. The change in protein composition accompanying hydrolysis also allows the spatial distribution of allergenic proteins to be estimated in the latex. Concurrently, the latex stability increased with the storage duration, as measured by the latex's mechanical stability time (MST) and the zeta potential of the latex particles. As monitored by AFM, the surface roughness of the NR latex film increased markedly during extended storage compared with that of the DPNR latex, which remained smooth. These results underscore the pivotal role of ammonia in bolstering NR latex stability brought on by protein hydrolysis, which greatly impacts latex film's formation behavior. NR latex stability underpins the quality of latex-dipped goods during manufacturing, particularly those for medical gloves.

Quercetin improves and protects Calu-3 airway epithelial barrier function

Introduction: In light of the impact of airway barrier leaks in COVID-19 and the significance of vitamin D in COVID-19 outcomes, including airway barrier protection, we investigated whether the very common dietary flavonoid quercetin could also be efficacious in supporting airway barrier function.Methods: To address this question, we utilized the widely used airway epithelial cell culture model, Calu-3.Results: We observed that treating Calu-3 cell layers with quercetin increased transepithelial electrical resistance while simultaneously reducing transepithelial leaks of 14C-D-mannitol (Jm) and 14C-inulin. The effects of quercetin were concentration-dependent and exhibited a biphasic time course. These effects of quercetin occurred with changes in tight junctional protein composition as well as a partial inhibition of cell replication that resulted in decreased linear junctional density. Both of these effects potentially contribute to improved barrier function. Quercetin was equally effective in reducing the barrier compromise caused by the pro-inflammatory cytokine TNF-α, an action that seemed to derive, in part, from reducing the elevation of ERK 1/2 caused by TNF-α.Discussion: Quercetin improved Calu-3 barrier function and reduced TNF-α-induced barrier compromise, mediated in part by changes in the tight junctional complex.

HIV-1 Nef Changes the Proteome of T Cells Extracellular Vesicles Depleting IFITMs and Other Antiviral Factors

Extracellular vesicles (EVs) are biomolecule carriers for intercellular communication in health and disease. Nef is a HIV virulence factor that is released from cells within EVs and is present in plasma EVs of HIV-1 infected individuals. We performed a quantitative proteomic analysis to fully characterize the Nef-induced changes in protein composition of T cell-derived EVs and identify novel host targets of HIV. Several proteins with well-described roles in infection or not previously associated with HIV pathogenesis were specifically modulated by Nef in EVs. Among the downregulated proteins are the interferon-induced transmembrane 1, 2, and 3 (IFITM1-3) proteins, broad-spectrum antiviral factors known to be cell-to-cell transferable by EVs. We demonstrate that Nef depletes IFITM1-3 from EVs by excluding these proteins from the plasma membrane and lipid rafts, which are sites of EVs biogenesis in T cells. Our data establish Nef as a modulator of EVs' global protein content and as an HIV factor that antagonizes IFITMs.

Impact of Different Light Conditions on the Nitrogen, Protein, Colour, Total Phenolic Content and Amino Acid Profiles of Cultured Palmaria palmata

The impact of different light conditions during culture on the nitrogen, protein, colour, total phenolic content (TPC) and amino acid profile of Palmaria palmata biomass was investigated. P. palmata was cultured using different light regimes, i.e., white (1 and 2), red, blue and green over 12 days. A significant decrease (p < 0.05) in total nitrogen (TN), non-protein nitrogen (NPN) and protein nitrogen (PN) was observed on day 6 while an increase was observed on day 12 in P. palmata samples cultured under blue light. The protein content (nitrogen conversion factor of 4.7) of the initial sample on day 0 was 15.0% (w/w) dw whereas a maximum protein content of 16.7% (w/w) was obtained during exposure to blue light following 12 days culture, corresponding to an 11.2% increase in protein content. Electrophoretic along with amino acid profile and score analyses showed light-related changes in protein composition. The lighting regime used during culture also influenced the colour parameters (lightness L*, redness a*, yellowness b* and colour difference ΔE) of milled algal biomass along with the TPC. Judicious selection of lighting regime during culture may allow the targeted production of sustainable high-quality proteins from P. palmata.

From grain to malt: Tracking changes of ultra-low-gluten barley storage proteins after malting

Barley (Hordeum vulgare L.) is a major cereal crop produced globally. Hordeins, the major storage proteins in barley, can trigger immune responses leading to celiac disease or symptoms associated with food allergy. Here, proteomics approaches were employed to investigate the proteome level changes of grain and malt from the malting barley cultivar, Sloop, and single-, double- and triple hordein-reduced lines. The triple hordein-reduced line is an ultra-low gluten barley cultivar, Kebari®. Using discovery proteomics, 2,688 and 3,034 proteins in the barley and malt samples were detected respectively. Through the application of targeted proteomics, a significant reduction in the quantity of B-, D-, and γ-hordeins, as well as avenin-like proteins, was observed in the ultra-low gluten malt sample. A compensation mechanism was observed evidenced by increased biosynthesis of seed storage globulins, specifically vicilin-like globulins. Overall, this study has provided insights into protein compositional changes after malting in celiac-friendly barley varieties.

Proteomic and Phenotypic Studies of Mycoplasma pneumoniae Revealed Macrolide-Resistant Mutation (A2063G) Associated Changes in Protein Composition and Pathogenicity of Type I Strains.

Mycoplasma pneumoniae (MP) is an important respiratory pathogen, the prevalence of macrolide-resistant MP (mainly containing A2063G mutation in 23S rRNA) increased in recent years. Epidemiological studies suggest a higher prevalence of type I resistant (IR) strains than corresponding sensitive (IS/IIS) strains, but not type II resistant (IIR) strains. Here, we aimed to analyze the factors underlying the altered prevalence of IR strains. First, proteomic analyses exhibit the protein compositions were type specific, while more differential proteins were detected between IS and IR (227) than IIS and IIR strains (81). mRNA level detection suggested posttranscriptional regulation of these differential proteins. Differential protein-related phenotypic changes were also detected: (i) P1 abundance was different between genotypes (I < II, IR < IS), the adhesion of MPs showed accordance to P1 abundance within IS and IIS strains; (ii) type I, especially IR, strains had a higher proliferation rate, which is potentially associated with differential proteins participating in glycolysis and one carbon pool metabolisms; (iii) A549 cells infected with IR strains had lower activity of caspase-3 and higher levels IL-8, but the differences were not significant between groups (P > 0.05). Correlations of P1 abundance to caspase-3 activity and proliferation rate to the level of IL-8 were obtained. These results suggest changes in protein composition influenced the pathogenicity of MP, especially in IR strains, which may impact the prevalence of MP strains of different genotypes. IMPORTANCE The prevalence of macrolide-resistant MPs increased the difficulty in treatment of MP infections and posed potential threats to children's health. Epidemiological studies showed a high prevalence of IR-resistant strains (mainly A2063G in 23S rRNA) in these years. However, the trigger mechanisms for this phenomenon are not clear. In this paper, proteomic and phenotypic studies suggest that IR strains have reduced levels of multiple adhesion proteins and increased proliferation rate, which may lead to higher transmission rate of IR strains in the population. This suggests that we should pay attention to the prevalence of IR strains.

Proteolytic Homeostasis in the Tissue of the Spleen and the Heart of Rats Injected with the Venom of Vipera berus berus and Vipera berus nikolskii

Species of the subfamily Viperinae, known to be Old World vipers, Vipera berus berus and Vipera berus nikolskii, are highly distributed in Europe and show their venomous effect, leading to proteolysis, thrombocytopenia, neurotoxicity, and haemorrhage state in susceptible organism. A shift in the protein balance underlies the envenoming in the targeted organs and in the whole organism. Thus, a study of the influence of the V. berus berus and V. berus nikolskii venoms on the protein systems of the spleen and the heart was conducted in order to single out the impact of toxins on the metabolic pathways in the targeted organs. The study included an investigation of the amount of total proteins, and their redistribution and connection with toxicity. The results prove the assumption of proteolytic activation and the emergence of the toxicity state, showing a decrease in the level of proteins, changes in protein composition, redistribution of enzymatic profiles, and an increased level of low molecular weight molecules.

Pseudomonas aeruginosa responds to altered membrane phospholipid composition by adjusting the production of two-component systems, proteases and iron uptake proteins

Membrane protein and phospholipid (PL) composition changes in response to environmental cues and during infections. To achieve these, bacteria use adaptation mechanisms involving covalent modification and remodelling of the acyl chain length of PLs. However, little is known about bacterial pathways regulated by PLs. Here, we investigated proteomic changes in the biofilm of P. aeruginosa phospholipase mutant (∆plaF) with altered membrane PL composition. The results revealed profound alterations in the abundance of many biofilm-related two-component systems (TCSs), including accumulation of PprAB, a key regulator of the transition to biofilm. Furthermore, a unique phosphorylation pattern of transcriptional regulators, transporters and metabolic enzymes, as well as differential production of several proteases, in ∆plaF, indicate that PlaF-mediated virulence adaptation involves complex transcriptional and posttranscriptional response. Moreover, proteomics and biochemical assays revealed the depletion of pyoverdine-mediated iron uptake pathway proteins in ∆plaF, while proteins from alternative iron-uptake systems were accumulated. These suggest that PlaF may function as a switch between different iron-acquisition pathways. The observation that PL-acyl chain modifying and PL synthesis enzymes were overproduced in ∆plaF reveals the interconnection of degradation, synthesis and modification of PLs for proper membrane homeostasis. Although the precise mechanism by which PlaF simultaneously affects multiple pathways remains to be elucidated, we suggest that alteration of PL composition in ∆plaF plays a role for the global adaptive response in P. aeruginosa mediated by TCSs and proteases. Our study revealed the global regulation of virulence and biofilm by PlaF and suggests that targeting this enzyme may have therapeutic potential.

Bone Marrow Mesenchymal Stem Cells Derived from Juvenile Macaques Reversed the Serum Protein Expression Profile in Aged Macaques.

The aim of the study was to reveal the changes in serum protein composition and content in macaques during the process of ageing, and explore the effect of bone marrow mesenchymal stem cell (BMMSC) on the serum protein expression profile in elderly macaques. Naturally ageing macaques were assessed according to age. BMMSCs were intravenously infused into aged macaques. In addition, peripheral blood was collected to obtain serum for dataindependent acquisition (DIA) protein sequencing to identify aging-related indicators. One hundred eighty days after macaques received BMMSC treatment, haemoxylin and eosin (HE) staining was performed to observe the morphology and structure of aortic arches. Compared to infant and young control macaques, aged macaques showed erythema on the face, dry skin, reduced amounts of hair on the head and back, and paleness. Cultured BMMSCs from the 4th passage (P4 BMMSCs) were grown in accordance with standards used to culture mesenchymal stem cells. After BMMSC treatment, the assessed aortic arches showed no calcium salt deposition or cell necrosis, and the characteristics of the serum protein expression profile tended to be similar to that of the infant and young groups, with the expression of 41 proteins upregulated with age and that of 30 proteins downregulated with age but upregulated after BMMSC treatment. Moreover, we identified 44 significantly differentially expressed proteins between the aged model and treatment groups; 11 of the upregulated proteins were related to vascular ageing, neuronal ageing and haematopoiesis, and 33 of the downregulated proteins were associated with neuronal ageing, cardiovascular disease, and tumours. Interestingly, S100 expression in serum was significantly decreased, COMP expression was significantly increased, NKAP expression reappeared, and LCN2, CSF1R, CORO1C, CSTB and RSU-1 expression disappeared after BMMSC treatment. BMMSCs can reverse ageing-related serum protein expression.

Influence of storage time on protein composition and simulated digestion of UHT milk and centrifugation presterilized UHT milk in vitro

The centrifugation presterilizing UHT (C-UHT) sterilization method removes 90% of the microorganism and somatic cells from raw milk using high-speed centrifugation following UHT treatment. This study aimed to study the changes in protein composition and plasmin in the UHT and C-UHT milk. The digestive characteristics, composition, and peptide spectrum of milk protein sterilized with the 2 technologies were studied using a dynamic digestive system of a simulated human stomach. The Pierce bicinchoninic acid assay, laser scanning confocal microscope, liquid chromatography-tandem mass spectrometry, and AA analysis were used to study the digestive fluid at different time points of gastric digestion in vitro. The results demonstrated that C-UHT milk had considerably higher protein degradation than UHT milk. Different processes resulted during the cleavage of milk proteins at different sites during digestion, resulting in different derived peptides. The results showed there was no significant effect of UHT and C-UHT on the peptide spectrum of milk proteins, but C-UHT could release relatively more bioactive peptides and free AA.

Optimization of Extrusion Treatments, Quality Assessments, and Kinetics Degradation of Enzyme Activities during Storage of Rice Bran.

Over the years, extrusion has been a multi-step thermal technique that has proven to be the most effective process to stabilize rice bran (RB). This study aimed to investigate the effects of extrusion treatment and temperature (15, 25, and 40 °C) on the storage stability, lipid oxidation, peroxidase, and peroxide values, free fatty acids, fatty acid composition, and protein variations of RB over 60 days. The study offers novel insights into the changes in RB's protein and amino acid compositions during extrusion and storage, which has not been extensively explored in prior research. After extrusion processing, peroxidase activity (POD) and lipase activity (LPS) were significantly reduced. However, peroxide value (PV), free fatty acids (FFA), and malondialdehyde content (MDA) observed a significantly increased by 0.64 mEqO2/kg, 8.3 mg/100 g, and 0.0005 μmol/L respectively. The storage stability of RB after extrusion shows that the POD, LPS, FFA, PV, and MDA were positively correlated with storage duration and temperature. The oleic acid/linoleic acid ratio in processed RB by extrusion had no significant changes during storage. The total and essential/non-essential amino acid ratios showed a downward trend of 5.26% and 8.76%, respectively. The first-order kinetics was the best-fitting model to describe the enzymatic inactivation and degradation of extruded RB during storage. The extrusion treatment did not affect the crude protein and the essential subunits of protein. Overall, the optimized extrusion procedure exhibited promising results in stabilizing the RB.

Multifrequency dielectric mapping of fixed mice colon tissues in cell culture media via scanning electrochemical microscopy.

Alternating current scanning electrochemical microscopy (AC-SECM) is a powerful tool for characterizing the electrochemical reactivity of surfaces. Here, perturbation in the sample is induced by the alternating current and altered local potential is measured by the SECM probe. This technique has been used to investigate many exotic a range of biological interfaces including live cells and tissues, as well as the corrosive degradation of various metallic surfaces, etc. In principle, AC-SECM imaging is derived from electrochemical impedance spectroscopy (EIS) which has been used for a century to describe interfacial and diffusive behaviour of molecules in solution or on a surface. Increasingly bioimpedance centric medical devices have become an important tool to detect evolution of tissue biochemistry. Predictive implications of measuring electrochemical changes within a tissue is one of the core concepts in developing minimally invasive and smart medical devices. In this study, cross sections of mice colon tissue were used for AC-SECM imaging. A 10 micron sized platinum probe was used for two-dimensional (2D) tan δ mapping of histological sections at a frequency of 10kHz, Thereafter, multifrequency scans were performed at 100Hz, 10kHz, 300kHz, and 900kHz. Loss tangent (tan δ) mapping of mice colon revealed microscale regions within a tissue possessing a discrete tan δ signature. This tan map may be an immediate measure of physiological conditions in biological tissues. Multifrequency scans highlight subtle changes in protein or lipid composition as a function of frequency which was recorded as loss tangent maps. Impedance profile at different frequencies could also be used to identify optimal contrast for imaging and extracting the electrochemical signature specific for a tissue and its electrolyte.

Characterization of the improved functionality in soybean protein-proanthocyanidins conjugates prepared by the alkali treatment

This study investigated the changes in functionality, structure, and protein composition in soybean protein-proanthocyanidins (SPI-PC) conjugates prepared by the alkali method with dose-ranging PC molecular grafting (0.5, 1, 2 mg/mL). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that more >180 kDa polymers were formed in SPI-PC conjugates with the increase of PC levels. The structural analysis showed that conjugation to PC altered the molecular structure of SPI, appearing the protein-unfolding with PC level-dependent. Liquid chromatography tandem MS analysis (LC/MS-MS) demonstrated that the composition of protein and peptides, as well as allergen and epitopes, were changed after SPI was conjugated with PC. The antioxidant activity of SPI-PC conjugates was increased with the increase of PC levels because high PC contents caused more hydroxyl groups in polyphenols to introduce into the SPI. Emulsifying properties were increased with the increase of PC levels, probably resulting from the unfolding of proteins, increase of random coil and decrease of surface hydrophobicity. Western-bolt and enzyme-linked immunosorbent assay (ELISA) showed a reduced IgE binding capacity in SPI-PC conjugates with PC level-dependent, which was attributed to the reduction of major soybean allergens and the masking or destruction of epitopes induced by structural changes. However, the foaming properties varied from PC levels. Overall, this study demonstrated that SPI-PC conjugates formed using the alkali method have a potential application as hypoallergenic soybean products or functional ingredients in foods. In future studies, suitable reactions (e.g., PC levels) are required to be investigated in SPI-PC conjugates for balancing various functional properties. • PC increased emulsifying, foaming and antioxidant properties in SPI. • The IgE binding capacity in SPI was reduced with the increase in PC levels. • Protein modifications and structural changes in SPI varied from PC levels. • Suitable PC levels are required to balance various functionality in conjugates.

Epitranscriptome changes triggered by ammonium nutrition regulate the proteome response of maritime pine roots.

Epitranscriptome constitutes a gene expression checkpoint in all living organisms. Nitrogen is an essential element for plant growth and development that influences gene expression at different levels such as epigenome, transcriptome, proteome, and metabolome. Therefore, our hypothesis is that changes in the epitranscriptome may regulate nitrogen metabolism. In this study, epitranscriptomic modifications caused by ammonium nutrition were monitored in maritime pine roots using Oxford Nanopore Technology. Transcriptomic responses mainly affected transcripts involved in nitrogen and carbon metabolism, defense, hormone synthesis/signaling, and translation. Global detection of epitranscriptomic marks was performed to evaluate this posttranscriptional mechanism in un/treated seedlings. Increased N6-methyladenosine (m6A) deposition in the 3'-UTR was observed in response to ammonium, which seems to be correlated with poly(A) lengths and changes in the relative abundance of the corresponding proteins. The results showed that m6A deposition and its dynamics seem to be important regulators of translation under ammonium nutrition. These findings suggest that protein translation is finely regulated through epitranscriptomic marks likely by changes in mRNA poly(A) length, transcript abundance and ribosome protein composition. An integration of multiomics data suggests that the epitranscriptome modulates responses to nutritional, developmental and environmental changes through buffering, filtering, and focusing the final products of gene expression.

Analyzing a Saturation Effect of Nitrogen Fertilization on Baking Volume and Grain Protein Concentration in Wheat

Some wheat cultivars show a linear relationship between grain protein concentration (GPC) and baking volume, but others display a saturation curve. Such a saturation curve could be general, but in some cultivars it might only appear at GPC &gt; 17%. However, such GPC is mostly not achieved in the field. Pot experiments with high nitrogen application reliably result in GPC &gt; 17%. In a pot experiment with a high (N1) and an excessive N level (N2) and four cultivars (Akteur, Arnold, Discus and Hystar), the change in grain protein composition and the relationship between different protein fractions and baking volume at GPC &gt; 17% was investigated. GPC ranged from 17 to 24% and mean nitrogen content per grain from 1.2 to 1.8 mg. The N2 treatment increased GPC and mean nitrogen content per grain in the Akteur and Discus cultivar, but not in Arnold and Hystar. N2 increased concentration of gliadin by 10 to 34% and glutenin macropolymer (GMP) in all cultivars by 12 to 73%. Glutenin concentration was increased by N2 in Akteur and Discus (19 to 36%), but was decreased by N2 in the Arnold and Hystar cultivar. Baking volume was moderately increased by N2 in all cultivars by 6 to 9% and correlated significantly with most glutenin fractions in the Akteur and Discus cultivar, with GMP in Arnold and with HMW-GS to LMW-GS ratio in Hystar. Thus, specific effects on grain protein by N2 were responsible for the increased baking volume in each cultivar. However, as gliadin and its sub-fractions hardly correlated with baking volume, a positive effect of increasing gliadin proteins on baking quality was not obvious.

PH-dependent sedimentation and protein interactions in ultra-high-temperature-treated sheep skim milk

Sheep milk is considered unstable to UHT processing, but the instability mechanism has not been investigated. This study assessed the effect of UHT treatment (140°C/5 s) and milk pH values from 6.6 to 7.0 on the physical properties of sheep skim milk (SSM), including heat coagulation time, particle size, sedimentation, ionic calcium level, and changes in protein composition. Significant amounts of sediment were found in UHT-treated SSM at the natural pH (∼6.6) and pH 7.0, whereas lower amounts of sediment were observed at pH values of 6.7 to 6.9. The proteins in the sediment were mainly κ-casein (CN)-depleted casein micelles with low levels of whey proteins regardless of the pH. Both the pH and the ionic calcium level of the SSM at all pH values decreased after UHT treatment. The dissociation levels of κ-, β-, and αS2-CN increased with increasing pH of the SSM before and after heating. The protein content, ionic calcium level, and dissociation level of κ-CN were higher in the SSM than values reported previously in cow skim milk. These differences may contribute to the high amounts of sediment in the UHT-treated SSM at natural pH (∼6.6). Significantly higher levels of κ-, β-, and αS2-CN were detected in the serum phase after heating the SSM at pH 7.0, suggesting that less κ-CN was attached to the casein micelles and that more internal structures of the casein micelles may have been exposed during heating. This could, in turn, have destabilized the casein micelles, resulting in the formation of protein aggregates and high amounts of sediment after UHT treatment of the SSM at pH 7.0.

Evaluation of the Thermal Processes on Changing the Phenotypic Characteristics of Escherichia coli Strains from Ice Cream Compared to Non-Pasteurized Milk

Escherichia coli (E. coli) is shocked by various temperature processes in milk, which forces the organism to make proteins as a result of changes in the synthesis of enzymes that might give the strain special characteristics. The purpose of this study was to investigate the effects of the heat shock factor on changing the results of biochemical and molecular tests among E. coli strains obtained from ice cream and non-pasteurized milk when compared to a reference strain from the American-type culture collection (ATCC) in order to determine the phenotypic variation caused by the temperature conditions of the manufacturing process. Furthermore, isolates with characteristics similar to E. coli were discovered, but they were not E. coli and caused some ambiguity. To test the E. coli contamination of traditional and industrial ice cream, 82 samples were chosen at random. SDS-PAGE and 16S rDNA sequencing were carried out, as well as phenotypic testing. Isolated strains did not exactly match the reference strain. The results of biochemical testing and protein analysis revealed that the isolates were diverse. Samples E. coli phenons were classified. In the electrophoresis, the ice cream strain had two protein bands in the 20.75 and 23.59 kDa ranges that were distinct from the reference strain. These isolates appear to experience alterations in enzyme characteristics and structural proteins as a result of being exposed to various temperature conditions, such as pasteurization and frigidity. When compared to the reference strain, the calculated similarity percentage of the elicited isolate varied from 60 to 70%. The electrophoretic patterns of E. coli isolated elicited from milk samples differed from E. coli isolated obtained from the ice cream. The distinctions were in the intensity or position of the bands. The results also revealed that when isolates are subjected to thermal stresses, they exhibit a pattern similar to that of ice cream isolates. These considerations are made because a change in protein composition might result in a change in biochemical features, resulting in uncertainty in its identification. Sequences revealed that the sequences were related to E. coli 16S rDNA, despite differences in phenotypic and electrophoretic features between the isolated bacteria and the reference strain E. coli ATCC 25922. Our findings revealed that 16S rDNA could potentially be used to instantly implement an appropriate preventive measure for the purpose of identifying this type of bacteria and avoid some ambiguity.