Protein Bands Research Articles

SARS-CoV-2 multi-epitope subunit vaccine proof-of-concept derived from the insilico study with protein expression in Escherichia coli BL21

The protein subunit vaccine is the most considerably developed SARS-CoV-2 vaccine, according to the WHO vaccine tracker in 2023. The acceleration of vaccine development in 2 years of eradicating the COVID-19 pandemic is attainable due to the role of bioinformatics. This paper aims to evaluate strategies for developing multi-epitope SARS-CoV-2 recombinant vaccines in silico with high protein expression in Escherichia coli vector plasmid. The study was conducted by analysing SARS-CoV-2 epitopes using immunoinformatic tools provided by IEDB, codon optimisation, rare codon analysis, plasmid design, and ribosomal binding site (RBS) analysis were analysed using RNA structure 6.4, gene cloning by E. coli DH5α and protein expression by E. coli BL21. Each epitope peptide candidate was linked to a flexible linker sequence (GGGGS). GelAnalyzer 19.1 was utilised to determine the protein band of SDS-PAGE. The immunoinformatic study obtained a multi-epitope of the recombinant SARS-COV-2 vaccine with 7 epitopes for HLA-I allele candidates and 4 for HLA-II. It is demonstrated that the candidate vaccine protein was successfully cloned in E. coli DH5α and expressed in E. coli BL21. The result of this study will benefit the development of a SARS-CoV-2 protein recombinant vaccine that is safe, affordable, and efficient to induce T-cell response.

Analysis of serum peptidome profiles of non-metastatic and metastatic feline mammary carcinoma using liquid chromatography-tandem mass spectrometry

BackgroundFeline mammary carcinoma (FMC) is a common aggressive and highly metastatic cancer affecting female cats. Early detection is essential for preventing local and distant metastasis, thereby improving overall survival rates. While acquiring molecular data before surgery offers significant potential benefits, the current protein biomarkers for monitoring disease progression in non-metastatic FMC (NmFMC) and metastatic FMC (mFMC) are limited. The objective of this study was to investigate the serum peptidome profiles of NmFMC and mFMC using liquid chromatography-tandem mass spectrometry. A cross-sectional study was conducted to compare serum peptidome profiles in 13 NmFMC, 23 mFMC and 18 healthy cats. The liquid chromatography-tandem mass spectrometry analysis was performed on non-trypsinized samples.ResultsOut of a total of 8284 expressed proteins observed, several proteins were found to be associated with human breast cancer. In NmFMC, distinctive protein expressions encompassed double-stranded RNA-binding protein Staufen homolog 2 (STAU2), associated with cell proliferation, along with bromodomain adjacent to zinc finger domain 2A (BAZ2A) and gamma-aminobutyric acid type A receptor subunit epsilon (GABRE), identified as potential treatment targets. Paradoxically, positive prognostic markers emerged, such as complement C1q like 3 (C1QL3) and erythrocyte membrane protein band 4.1 (EPB41 or 4.1R). Within the mFMC group, overexpressed proteins associated with poor prognosis were exhibited, including B-cell lymphoma 6 transcription repressor (BCL6), thioredoxin reductase 3 (TXNRD3) and ceruloplasmin (CP). Meanwhile, the presence of POU class 5 homeobox (POU5F1 or OCT4) and laminin subunit alpha 1 (LAMA1), reported as metastatic biomarkers, was noted.ConclusionThe presence of both pro- and anti-proliferative proteins was observed, potentially indicating a distinctive characteristic of NmFMC. Conversely, proteins associated with poor prognosis and metastasis were noted in the mFMC group.

Presence of low-energy chlorophylls d in photosystem I trimer and monomer cores isolated from Acaryochloris sp. NBRC 102871.

Acaryochloris species belong to a special category of cyanobacteria possessing chlorophyll (Chl) d. One of the photosynthetic characteristics of Acaryochloris marina MBIC11017 is that the absorption spectra of photosystem I (PSI) showed almost no bands and shoulders of low-energy Chls d over 740nm. In contrast, the absorption spectra of other Acaryochloris species showed a shoulder around 740nm, suggesting that low-energy Chls d within PSI are diversified among Acaryochloris species. In this study, we purified PSI trimer and monomer cores from Acaryochloris sp. NBRC 102871 and examined their protein and pigment compositions and spectral properties. The protein bands and pigment compositions of the PSI trimer and monomer of NBRC102871 were virtually identical to those of MBIC11017. The absorption spectra of the NBRC102871 PSIs exhibited a shoulder around 740nm, whereas the fluorescence spectra of PSI trimer and monomer displayed maximum peaks at 754 and 767nm, respectively. These spectral properties were different from those of MBIC11017, indicating the presence of low-energy Chls d within the NBRC102871 PSIs. Moreover, we analyzed the NBRC102871 genome to identify amino acid sequences of PSI proteins and compared them with those of the A. marina MBIC11017 and MBIC10699 strains whose genomes are available. The results showed that some of the sequences in NBRC102871 were distinct from those in MBIC11017 and MBIC10699. These findings provide insights into the variety of low-energy Chls d with respect to the protein environments of PSI cores among the three Acaryochloris strains.

Recovery of Proteases and Protease Inhibitors from Ganoderma spp. Cultivated in Amazonian Lignocellulose Wastes.

Ganoderma spp. are a great source of bioactive molecules. The production and recovery of bioactive molecules vary according to strain, growth substrate, and extraction solution. Variations in protease and their inhibitors in basidiomata from a commercial strain (G. lingzhi) and an Amazonian isolate (Ganoderma sp.) cultivated in Amazonian lignocellulosic wastes and extracted with different solutions are plausible and were investigated in our study. Basidiomata from cultivation in substrates based on açaí seed, guaruba-cedro sawdust and three lots of marupá sawdust were submitted to extraction in water, Tris-HCl, and sodium phosphate. Protein content, proteases, and protease inhibitors were estimated through different assays. The samples were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR). Tris-HCl provided higher protein extraction from Ganoderma sp. and higher caseinolytic, gelatinolytic, and fibrinolytic activity for G. lingzhi cultivated in açaí. Water extracts of Ganoderma sp., in general, exhibited higher trypsin and papain inhibitor activities compared to G. lingzhi. Extracts in Tris-HCl and sodium phosphate showed more intense protein bands in SDS-- PAGE, highlighting bands of molecular weights around 100, 50, and 30 kDa. FTIR spectra showed patterns for proteins in all extracts, with variation in transmittance according to substrate and extractor. Water extract from Amazonian Ganoderma sp. cultivated in marupá wastes are promising as a source of protease inhibitors, while the Tris-HCL extract of G. lingzhi from açaí cultivation stands out as a source of proteases with fibrinolytic, caseinolytic, and gelatinolytic activities.

Valorization of agro-industrial residues using Aspergillus heteromorphus URM0269 for protease production: Characterization and purification

This study aimed to produce, characterize and purify a protease from Aspergillus heteromorphus URM0269. After production by solid fermentation of wheat bran performed according to a central composite design, protease was characterized in terms of biochemical, kinetic, and thermodynamic parameters for further purification by chromatography. Proteolytic activity achieved a maximum value of 57.43 U/mL using 7.8 g of wheat bran with 40 % moisture. Protease displayed high stability in the pH and temperature ranges of 5.0–10.0 and 20–30 °C, respectively, and acted optimally at pH 7.0 and 50 °C. The enzyme, characterized as a serine protease, followed Michaelis-Menten kinetics with a maximum reaction rate of 140.0 U/mL and Michaelis constant of 11.6 mg/mL. Thermodynamic activation parameters, namely activation Gibbs free energy (69.79 kJ/mol), enthalpy (5.86 kJ/mol), and entropy (−214.39 J/mol.K) of the hydrolysis reaction, corroborated with kinetic modeling showing high affinity for azocasein. However, thermodynamic parameters suggested a reversible mechanism of unfolding. Purification by chromatography yielded a protease purification factor of 7.2, and SDS-PAGE revealed one protein band with a molecular mass of 14.7 kDa. Circular dichroism demonstrated a secondary structure made up of 45.6 % α-helices. These results show the great potential of this protease for future use in the industrial area.

Unpicking the changes in content, structural properties and in vitro digestibility of pigeon pea seed proteins as influenced by imbibition and heating in sodium bicarbonate

BackgroundPigeon pea proteins are underutilised mainly because of the long cooking time of the legume seeds. Traditionally, this is mitigated by cooking pigeon pea seeds in sodium bicarbonate (NaHCO3). This study investigated whether, and to what degree, directly cooking pigeon pea seeds in water (CS0W) or NaHCO3 (CS0N), soaking in water or NaHCO3 for 4 h (CS4W and CS4N, respectively) or for 24 h (CS24W and CS24N, respectively) before cooking in water or NaHCO3 influenced the protein content, structural property and in vitro digestibility. ResultsIn vitro protein digestibility of pigeon pea seed flour was lowest in the raw seeds and increased with CS0W<CS4W<S24CW and CS0N<CS4N<S24CN. The α-helix content of the samples also decreased in the same order. SDS-PAGE showed ∼90% decrease in the 7S vicilin subunits (65 and 47 kDa) in CS24N and a corresponding 4-fold increase in protein bands at 10–15 kDa compared to raw seeds. NaHCO3 treatments decreased the protein isolate yields due to protein losses (up to 7.5-folds) in the cooking fluids. ConclusionThe findings showed that NaHCO3 improved protein digestibility by degrading the higher molecular weight proteins, but also increased protein diffusion into the cooking fluid. Therefore, the traditional method of cooking pigeon pea seeds needs optimization to maximize the protein accessibility.

Serum proteinogram of gilthead seabream (Sparus aurata) and European sea bass (Dicentrarchus labrax) as a new useful approach for detecting loss of haemostasis

Proteinograms, a semiquantitative analytical method that separates proteins into multiple bands, have not been explored in teleosts for diagnostic or prognostic purposes. This study aimed to establish reference values for proteinograms in the serum of gilthead seabream (Sparus aurata) and European sea bass (Dicentrarchus labrax), two important farmed fish species in the Mediterranean region. Serum proteins were studied using SDS-PAGE, electropherogram, and HPLC-mass spectrometry. SDS-PAGE analysis revealed four major bands of proteins around 11, 25, 70, and 100 kDa in the serum of gilthead seabream and European sea bass. Electropherogram results showed that a protein with a molecular weight of 76.8 kDa was the most abundant protein in the serum of gilthead seabream, while a peak of 75.5 kDa was the most abundant in European sea bass. HPLC-mass spectrometry detected 87 proteins and 119 proteins in the serum of gilthead seabream and European sea bass, respectively, including α1-globulins, α2-globulins, β-globulins, and γ-globulins. Notably, the albumin sequence was not detected in either of the two species. These results help to characterize the serum protein profile and to establish reference proteinograms for these two fish species. They also provide a basis for the development of novel approaches for the rapid detection of loss of haemostasis due to stress, health disorders or disease in farmed fish.

Anticancer Activities of Bromelain Hydrolysate of Soy Protein Against Breast Cancer Cells MCF-7

Soybeans contain proteins that have the potential to produce anticancer bioactive peptides. This study aims to determine the anticancer activity of soy protein hydrolysate against MCF-7 breast cancer cells. Soybean protein hydrolyzed by bromelain enzyme 0.5% (w/v) at the optimum temperature and pH for protein hydrolysis using the Bergmeyer and Grassl method. The degree of hydrolysis value of protein hydrolysate was determined by the Alder-Niesen method and the protein profile was analyzed by SDS-PAGE. The hydrolysate with the best degree of hydrolysis value was analyzed for anticancer activity against MCF-7 breast cancer cells by the Presto Blue assay method, and fractionation of protein hydrolysates by gel filtration chromatography (Sephadex G-15). The molecular weight of the peptide was characterized by LCMS/MS. Soy protein hydrolysis using 0.5% (w/v) bromelain enzyme was optimum at 65 ºC and pH 7.0 for 4 hours, with a hydrolysis degree value of 20.57%. The SDS-PAGE analysis showed that the protein hydrolysates had quite thick protein bands in the range of &lt;35 kDa with an IC50 value of 70.37 mg/mL. Based on the LCMS/MS results, the peptide from fractionation has a molecular weight of 5.133 kDa.

Analysis of CFTR mRNA and Protein in Peripheral Blood Mononuclear Cells via Quantitative Real-Time PCR and Western Blot.

The Cystic Fibrosis Conductance Transmembrane Regulator gene encodes for the CFTR ion channel, which is responsible for the transport of chloride and bicarbonate across the plasma membrane. Mutations in the gene result in impaired ion transport, subsequently leading to perturbed secretion in all exocrine glands and, therefore, the multi-organ disease cystic fibrosis (CF). In recent years, several studies have reported on CFTR expression in immune cells as demonstrated by immunofluorescence, flow cytometry, and immunoblotting. However, these data are mainly restricted to single-cell populations and show significant variation depending on the methodology used. Here, we investigated CFTR transcription and protein expression using standardized protocols in a comprehensive panel of immune cells. Methods: We applied a high-resolution Western blot protocol using a combination of highly specific monoclonal CFTR antibodies that have been optimized for the detection of CFTR in epithelial cells and healthy primary immune cell subpopulations sorted by flow cytometry and used immortalized cell lines as controls. The specificity of CFTR protein detection was controlled by peptide competition and enzymatic Peptide-N-Glycosidase-F (PNGase) digest. CFTR transcripts were analyzed using quantitative real-time PCR and normalized to the level of epithelial T84 cells as a reference. Results: CFTR mRNA expression could be shown for primary CD4+ T cells, NK cells, as well as differentiated THP-1 and Jurkat T cells. In contrast, we failed to detect CFTR transcripts for CD14+ monocytes and undifferentiated THP-1 cells, as well as for B cells and CD8+ T cells. Prominent immunoreactive bands were detectable by immunoblotting with the combination of four CFTR antibodies targeting different epitopes of the CFTR protein. However, in biosamples of non-epithelial origin, these CFTR-like protein bands could be unmasked as false positives through peptide competition or PNGase digest, meaning that the observed mRNA transcripts were not necessarily translated into CFTR proteins, which could be detected via immunoblotting. Our results confirm that mRNA expression in immune cells is many times lower than in that cells of epithelial origin. The immunoreactive signals in immune cells turned out to be false positives, and may be provoked by the presence of a high-affinity protein with a similar epitope. Non-specific binding (e.g., Fab-interaction with glycosyl branches) might also contribute to false positive signals. Our findings highlight the necessity of accurate controls, such as CFTR-negative cells, as well as peptide competition and glycolytic digest in order to identify genuine CFTR protein by immunoblotting. Our data suggest, furthermore, that CFTR protein expression data from techniques such as histology, for which the absence of a molecular weight or other independent control prevents the unmasking of false positive immunoreactive signals, must be interpreted carefully as well.

Assessment of Biochemical Changes of Four Aspergillus Species Grown on the Medium from Agricultural Wastes

Hazardous disposal of agricultural wastes (AW) has adverse environmental consequences, including water and air pollution and the potential for disease outbreaks. On the other hand, the utilization of AW represents a missed opportunity to harness a valuable economic resource. This study was conducted with the objective of utilizing a composite medium comprising agricultural waste to cultivate Aspergillus species and assessing its impact on the species' internal chemical composition compared to malt extract media (ME). Our findings demonstrate that the agricultural waste-based medium is abundant in essential nutrients, including soluble proteins and sugars, and is also enriched with a variety of secondary metabolites. Consequently, this Change in the growth medium induces changes in the physical characteristics of fungal biomass, such as color and texture, along with a high content of biomass proteins and secondary metabolites, including phenols, flavonoids, carotenoids, and antioxidants. The A. avenaceous gave the highest biomass (1.1412 ± 0.4 g), while the A. niger gave the highest value of proteins (16.06 ± 0.4 mg/g), phenols (33.37 ± 0.8 mg/g), flavonoids (4.84 ± 0.4 mg/g), carotenoids (1.131 ± 0.09 mg/g). A. carbonarius gave the highest value of antioxidants (IC50 = 0.28 ± 0.06 mg/mL). In contrast, using malt extract as a growth medium results in high carbohydrate and lipid production; A. flavus showed the highest value for fats (56.6 ± 0.9 mg/g), whereas A. carbonarius showed the highest value for sugars (167.1 ± 6.2 mg/g). Additionally, the malt extract medium contributed to low levels of secondary metabolites, which was offset by an increase in the protein bands of the fungal species. This research recommends the use of agricultural wastes to grow fungi species as an environmentally and economically important microbiological application.

Analysis of protein profile and reducing sugar content of cassava plants (Manihot esculenta Crantz) results of induced resistance with salicylic acid

Cassava (Manihot esculenta Crantz) is an important food commodity in Indonesia, and in the future this commodity will play an increasingly strategic role in people's lives and the country's economy. But until now there are still many obstacles, one of which is fusarium wilt disease. Fusarium wilt disease can be controlled by using superior cultivars that are resistant to the disease, by applying salicylic acid. The aim of this study was to analyze the protein profile and reducing sugar content in cassava plants after being induced with salicylic acid and compare them with controls. This research used a Completely Randomized Design (CRD) with one factor, namely the addition of salicylic acid consisting of 5 concentration levels, namely 0 ppm, 80 ppm, 100 ppm, 120 ppm, and 140 ppm. Each concentration was repeated 5 times. This research data is in the form of qualitative data and quantitative data. Qualitative data is presented in comparative descriptive form and presented in the form of photographs. Quantitative data was tabulated with different concentration factors and analyzed using Analysis of Variance then further tested with the Honestly Significant Difference Test (BNJ) at the 5% level. The research results showed that there was a missing protein band, namely a band at a molecular weight of 115, and there was also the appearance of a new band, namely a molecular weight of 85 kDa, as well as an increase in the reducing sugar content in cassava plants along with increasing salicylic acid concentrations.

Whole Genome Linkage and Association Analyses Identify DLG Associated Protein-1 as a Novel Positional and Biological Candidate Gene for Muscle Strength: The Long Life Family Study.

Grip strength is a robust indicator of overall health, is moderately heritable, and predicts longevity in older adults. Using genome-wide linkage analysis, we identified a novel locus on chromosome 18p (mega-basepair region: 3.4-4.0) linked to grip strength in 3755 individuals from 582 families aged 64 ± 12 years (range 30-110 years; 55% women). There were 26 families that contributed to the linkage peak (cumulative logarithm of the odds [LOD] score = 10.94), with 6 families (119 individuals) accounting for most of the linkage signal (LOD = 6.4). In these 6 families, using whole genome sequencing data, we performed association analyses between the 7312 single nucleotide (SNVs) and insertion deletion (INDELs) variants in the linkage region and grip strength. Models were adjusted for age, age2, sex, height, field center, and population substructure. We found significant associations between genetic variants (8 SNVs and 4 INDELs, p < 5×10-5) in the Disks Large-associated Protein 1 (DLGAP1) gene and grip strength. Haplotypes constructed using these variants explained up to 98.1% of the LOD score. Finally, RNAseq data showed that these variants were significantly associated with the expression of nearby Myosin Light Chain 12A (MYL12A), Structural Maintenance of Chromosomes Flexible Hinge Domain Containing 1 (SMCHD1), Erythrocyte Membrane Protein Band 4.1 Like 3 (EPB41L3) genes (p < .0004). The DLGAP1 gene plays an important role in the postsynaptic density of neurons; thus, it is both a novel positional and biological candidate gene for follow-up studies aimed at uncovering genetic determinants of muscle strength.

Kinetic studies on optimized extracellular laccase from Trichoderma harzianum PP389612 and its capabilities for azo dye removal

BackgroundAzo dyes represent a common textile dye preferred for its high stability on fabrics in various harsh conditions. Although these dyes pose high-risk levels for all biological forms, fungal laccase is known as a green catalyst for its ability to oxidize numerous dyes.MethodsTrichoderma isolates were identified and tested for laccase production. Laccase production was optimized using Plackett–Burman Design. Laccase molecular weight and the kinetic properties of the enzyme, including Km and Vmax, pH, temperature, and ionic strength, were detected. Azo dye removal efficiency by laccase enzyme was detected for Congo red, methylene blue, and methyl orange.ResultsEight out of nine Trichoderma isolates were laccase producers. Laccase production efficiency was optimized by the superior strain T. harzianum PP389612, increasing production from 1.6 to 2.89 U/ml. In SDS-PAGE, purified laccases appear as a single protein band with a molecular weight of 41.00 kDa. Km and Vmax values were 146.12 μmol guaiacol and 3.82 μmol guaiacol/min. Its activity was stable in the pH range of 5–7, with an optimum temperature range of 40 to 50 °C, optimum ionic strength of 50 mM NaCl, and thermostability properties up to 90 °C. The decolorization efficiency of laccase was increased by increasing the time and reached its maximum after 72 h. The highest efficiency was achieved in Congo red decolorization, which reached 99% after 72 h, followed by methylene blue at 72%, while methyl orange decolorization efficiency was 68.5%.ConclusionTrichoderma laccase can be used as an effective natural bio-agent for dye removal because it is stable and removes colors very well.

Controlling Botrytis gray mold in strawberry fruit by bioactive protein isolated from kidney bean

Due to the risks they pose to human and environmental health, there has been a strong push in recent times to reduce the use of chemical fungicides in phytopathogen treatment. In the present study, vicilin was isolated from kidney bean seeds and characterized by SDS-PAGE, zeta potential, and pH solubility curve. The α/ (85 KDa), α (70 KDa), and β (60 KDa) subunits were represented by three distinct protein bands in vicilin. The linear growth of B. cinerea was clearly reduced by 12.96%, 14.81%, 25.92%, 35.18%, and 40.73% in response to vicilin application at 50, 100, 200, 300, and 400 µg/ml, respectively. The scanning electron microscopy (SEM) of vicilin-exposed B. cinerea revealed hyphae enlargement and conidia distortion. Addition of vicilin from kidney bean seeds clearly reduced the disease incidence in a concentration-dependent manner (100, 200, and 400 µg/mL). The higher doses (400 µg/mL) of vicilin provided higher activity in decreasing the disease severity of the strawberry fruits. As a sustainable glycoprotein, vicilin, found in kidney bean seeds, can be used to combat postharvest fungal infections.

Characterization of Polyphenol Composition and Starch and Protein Structure in Brown Rice Flour, Black Rice Flour and Their Mixtures.

The study investigates the structural and chemical properties of brown rice flour (WRF), black rice flour (BRF) and their mixtures in ratios of 25%, 50% and 75% to provide reference information for the gluten-free bakery industry. BRF contains higher concentrations of proteins, lipids, total minerals, crude fiber, total polyphenols, proanthocyanidins and flavonoids than WRF. A higher amylose content in BRF than in WRF resulted in flour mixtures with slower starch digestion and a lower glycemic response depending on the BRF ratio added. Differences in the chemical composition of WRF and BRF led to improved composition of the flour mixtures depending on the BRF ratio. The presence of anthocyanidins and phenolic acids in higher concentrations in the BRF resulted in a red-blue color shift within the flour mixtures. The deconvoluted FTIR spectra showed a higher proportion of α-helixes in the amide I band of BRF proteins, indicating their tighter folding. An analysis of the FTIR spectra revealed a more compact starch structure in BRF than in WRF. By processing reflection spectra, nine optically active compound groups were distinguished in rice flour, the proportion in BRF being 83.02% higher than in WRF. Due to co-pigmentation, the bathochromic shift to higher wavelengths was expressed by the proanthocyanins and phenolic acids associated with the wavelengths 380 nm to 590 nm and at 695 nm. Anthocyanins, protein-tannin complexes, methylated anthocyanins and acylated anthocyanins, associated with wavelengths 619, 644 and 668 nm, exhibited a hypsochromic effect by shifting the wavelengths to lower values. This research represents a first step in the development of rice-based products with increased nutritional value and a lower glycemic index.

Antioxidant potential of three different Sea weeds from Visakhapatnam Coast, India

In the present study the antioxidant potential of three seaweeds Gracileria corticata (red algae), Ulva lactuca (green algae) and Sargassum ilicifolum (brown algae) were reported. SDS-PAGE analysis reveals that total of 23 different protein bands were observed of this 16, 14 and 17 bands in U. lactuca, S. ilicifolium and G. Corticata respectively. The chloroform, acetone and methanolic extracts of three algae showed the concentration dependent DPPH, FRAP, reducing power, H2O2 radical scavenging and metal chelating activities. The methanolic extracts of G. corticata have high levels of antioxidants with high DPPH, FRAP, reducing power activities and S. ilicifolum have high H2O2 radical scavenging, and metal chelating activities.G. corticata showed IC50 value for 902.09µg/ml for DPPH activity, 2274µg/ml for FRAP activity, 1082.11µg/ml for H2O2 radical scavenging. Among the three solvents tested, Methanol was found to be the most suitable solvent for the extraction of compounds.

Development of Drying Methodology for Intact Whole Buffalo Liver, Its Characterization, Shelf Life, and Evaluation of Palatability as Pet Treat

Globally, large quantity of animal byproducts is generated from the slaughter of food animals, but there is lack of research articles related to drying of these byproducts and its use as pet food. Therefore, this study was conducted with the aim of utilization of intact whole buffalo liver by drying for pet treat, evaluating its shelf life and palatability. The intact liver surface was superficially sliced, and the surface was pierced. Thereafter, the livers were pretreated in 3% sugar and 4% salt solution (1 : 3 w/v) for 3 h followed by microwaving for 4 min and hot air drying at 60°C for 40 h (designated as T2L). The livers which were dried the same as T2L except surface piercing were referred as T1L, whereas the livers dried only using hot air oven were referred as control (CL). The moisture and protein contents of the dried CL and T2L were found to be 28.46% and 14.29% and 43.85% and 52.76%, respectively. Sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) image of T2L revealed the presence of few low as well as high molecular weight protein bands which were absent in CL and T1L indicating a comparatively lower level of protein degradation in T2L. The shelf life of T1L and T2L samples based on microbiological and lipid oxidation analyses was found to be more than 60 days at 25 and 4°C. Palatability studies using dogs showed that all dried samples were highly palatable. Thus, it could be concluded that intact buffalo liver could be dried using surface slicing, with/without piercing followed by salt and sugar pretreatment, 4 min microwaving, and hot air drying at 60°C for 40 h. Future study should focus on the sensory properties such as aroma, texture, and flavor and sensory analysis of the dried liver by human.

Multifunctional hydrophobin-like protein (HFB-NJ1): A versatile solution for wastewater treatment

As wastewater contains a variety of contaminating bacteria and oily residues, there is an urgent need for environmentally safe bactericidal agents and surfactants which can be applied for wastewater treatment. The present study emphasizes on the potential of hydrophobin-like protein (HFB-NJ1) extracted from sporulating mycelia of Aspergillus sp. NJ1 for wastewater treatment. The purified HFB-NJ1, depicted the presence of one single protein band of molecular size approximately 11–12 kDa on silver-stained SDS-PAGE gel. HFB-NJ1 also presented properties such as surface modification of glass and stable emulsification of sunflower oil. HFB-NJ1 depicted exceptional antibacterial activity against bacterial pathogens such as Bacillus subtilis and Pseudomonas aeruginosa at low MIC of 0.5 μg/mL and 0.75 μg/mL respectively. Additionally, HFB-NJ1 depicted enhanced emulsification of various vegetable and petroleum-based oils (E24 > 80%). HFB-NJ1 effectively reduced gold ions, producing nanospheres with a size of 15.33 nm – a recognized antimicrobial agent. This study underscores the multifunctional attributes of HFB-NJ1, highlighting its efficacy in removing pathogenic bacteria, emulsifying organic compounds from wastewater, and demonstrating a reduction ability for nanoparticle synthesis.

Development of a glutamate-functionalized macroporous cation-exchange matrix for partial purification of lysozyme from chicken egg white

The polymeric monoliths, called cryogels, arouse interest due to their versatility and to be a good alternative for enzyme purification processes. Lysozyme is an enzyme that stands out for its antimicrobial activity and is being used as a preservative in the food industry. This work aimed to produce a new cationic exchange matrix functionalized with monosodium glutamate for partial lysozyme purification of egg white. The structural characteristics of the support delivered were evaluated, presenting an ion exchange capacity (IEC) of 362×10−3 mol Na+. kg−1dry cryogel, zero charge point (PCZ) around pH 6.0, and porosity more significant than 90 %. The FTIR and TGA analyses proved that the modification of the XRD matrix indicated the material's amorphous nature. The hydrodynamic studies showed asymmetry, indicating the pores' non-uniformity and low resistance to flow. The lysozyme was purified from egg white, and a purification factor of 15.11 was obtained. SDS-PAGE electrophoresis analysis indicated high purity of lysozyme, with no other protein bands appearing in the eluted solution, indicating that the adsorbent has high potential for application in lysozyme purification processes.

Utilizing outer membrane proteins (OMPs) as a vaccine to manage motile Aeromonas septicemia (MAS) in Nile tilapia (Oreochromis niloticus) with evaluating the vaccine immunoprotective effectiveness

Motile aeromonads are an opportunistic and heterogenic bacteria that cause motile Aeromonas septicemia (MAS) which is accompanied with variety of clinical signs and lesions in fish. It has been found that their outer membrane proteins (OMPs) are highly immunogenic and may be used as a vaccine candidate. The present study was undertaken to evaluate OMP protection against Aeromonas hydrophila (A. hydrophila) strain ATCC 7966 (A5 and A8) and A. hydrophila strain DSM 30,187 (A6). Pathogenicity tests revealed that out of 12 isolated strains, seven were highly pathogenic, three were moderate, and two were avirulent. SDS-PAGE analysis of OMPs showed several protein bands (7–22) of different molecular weights ranging from 14 to 346 kDa. The common protein band was 39 kDa. It seems that pathogenicity and the number of protein bands are related in the highly pathogenic strains. Challenged A. hydrophila for Nile tilapia (O. niloticus) previously actively immunized with OMPs (A4, A5, A6, A8, and A11) showed a significant increase in relative percent survival (RPS) ranging from 50 to 100%. Immunological parameters such as immunoglobulin M (IgM), antioxidants (SOD, GSH, and CAT), and nitric oxide (NO) and expression of pro-inflammatory cytokines, interleukin 1β (IL-1β), and tumor necrosis factor-α (TNFα) showed a significant increase in OMP-immunized fish. Challenged A. hydrophila for O. niloticus previously passively immunized with OMPs (A4, A5, A6, A8, and A11) in fish-raised sera showed a significant increase in RPS (20 to 100%) in the vaccinated groups. It is concluded that OMPs from Aeromonas spp. have been identified as suitable target for protection development, especially in the case of heterogenic bacteria.