High Protein Research Articles

Expression of Major Basic Protein and Endothelial Adhesion Molecules in Chronically Inflamed Mucosa of the Ethmoid Labyrinth.

Tissue remodeling, including dense eosinophil infiltration, is essential for forming inflammatory nasal polyps (NPs) and the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP). Toxic eosinophil major basic protein (MBP) damages the sinus mucosa epithelium and lamina propria, which initiates reparative processes leading to tissue remodeling. MBP specifically binds to BMK-13 antibodies allowing immunohistochemical (IHC) tissue staining for eosinophils. This study evaluated the association between NP stromal BMK-13 and endothelial adhesion molecule staining, and clinical parameters of NP patients compared to IHC expression and clinical parameters in subjects with healthy nasal mucosa. We included 30 patients with bilateral NPs who were selected for endoscopic ethmoidectomy. The control group was of 30 subjects with non-inflamed nasal mucosa but with middle turbinate aeration, chosen for surgery. All participants were clinically scored before surgery, according to quality of life (QoL) outcome and symptoms. The degree of disease extension on computed tomography scans of the paranasal sinuses was also evaluated. Tissue samples after surgery were IHC stained for BMK-13 and endothelial proliferation markers CD31 and CD34. Expression of BMK-13, CD31, and CD34 in tissues of NPs was higher than in healthy nasal mucosa. Positive correlations were observed between BMK-13 expression, impaired QoL, and radiologically assessed extension of inflammation in NP patients. Apart from the fact that the NP tissue has, as expected, more intense eosinophilic infiltration, the proliferation of blood vessels is more pronounced in the NP tissue than in the tissue of healthy nasal mucosa. Expression of MBP in the tissue of ethmoidal NPs could serve as a potential marker of the degree of expansion of CRSwNP and indicate the severity of the disease. None, because it was a cross-sectional study.

Advances in Protein-Ligand Binding Affinity Prediction via Deep Learning: A Comprehensive Study of Datasets, Data Preprocessing Techniques, and Model Architectures.

Drug discovery is a complex and expensive procedure involving several timely and costly phases through which new potential pharmaceutical compounds must pass to get approved. One of these critical steps is the identification and optimization of lead compounds, which has been made more accessible by the introduction of computational methods, including deep learning (DL) techniques. Diverse DL model architectures have been put forward to learn the vast landscape of interaction between proteins and ligands and predict their affinity, helping in the identification of lead compounds. This survey fills a gap in previous research by comprehensively analyzing the most commonly used datasets and discussing their quality and limitations. It also offers a comprehensive classification of the most recent DL methods in the context of protein-ligand binding affinity prediction (BAP), providing a fresh perspective on this evolving field. We thoroughly examine commonly used datasets for BAP and their inherent characteristics. Our exploration extends to various preprocessing steps and DL techniques, including graph neural networks, convolutional neural networks, and transformers, which are found in the literature. We conducted extensive literature research to ensure that the most recent deep learning approaches for BAP were included by the time of writing this manuscript. The systematic approach used for the present study highlighted inherent challenges to BAP via DL, such as data quality, model interpretability, and explainability, and proposed considerations for future research directions. We present valuable insights to accelerate the development of more effective and reliable DL models for BAP within the research community. The present study can considerably enhance future research on predicting affinity between protein and ligand molecules, hence further improving the overall drug development process.

Expanding the Toolset of Biomolecular NMR with Efficient and Cost-Effective 17O-Labeling via Bacterial Expression.

Oxygen plays a central role in biomolecular structures and functions, with 17O NMR emerging as a powerful tool for elucidating biomolecular properties. However, the low natural abundance of the NMR-active isotope, 17O (0.0373 %), presents a significant hurdle to its widespread application. Here, we introduce a rapid and cost-effective approach for amino acid-specific 17O-labeling of recombinant proteins. Using a common bacterial expression system and with a 30-minute rapid synthesis protocol of 17O-labeled amino acids via mechanochemical saponification, we have generated Leu- and Phe-specific 17O-labeled recombinant proteins derived from diverse organisms, including CrgA and FtsQ from Mycobacterium tuberculosis and E protein from SARS-CoV-2 virus, demonstrating the applicability of our technique for amino acids known to be isotopically labeled without scrambling. We have acquired magic-angle-spinning 17O NMR of these proteins to confirm the successful 17O labeling and illustrate the sensitivity of 17O NMR to the protein's local structural environments. Our work significantly broadens the accessibility of 17O-NMR, empowering researchers to delve deeper into protein biophysics and biochemistry. This approach opens new avenues for understanding cellular processes at the molecular level by providing an effective tool for investigating oxygen-related interactions and chemistry within biomolecules.

Ultrasonic treatment of emulsion gels with different soy protein-hemp protein composite ratios: Changes in structural and physicochemical properties

To improve the emulsion gel system of single soybean isolate protein (SPI) and to broaden the application field of hemp protein isolate (HPI), ultrasonic treatment and HPI were introduced to improve the properties of SPI emulsion gel and to explore the mechanism. The results showed that the gel strength (218.6 g) and water-holding capacity (86.24 %) of the emulsion gels were improved under ultrasonic treatments when the ratio of SPI:HPI was >6:4, and the reticulation structure of the gels was enhanced. When the ratio of SPI:HPI was <6:4, the gel structure was loose and formless. Ultrasonic treatment has a significant effect on the emulsion gel with the ratio of SPI:HPI was >6:4. Appropriate ultrasonic treatment (400 W) changed the protein structure, improved the rheological properties of emulsion gels to form the protein-oil-coated network structure. However, excessive ultrasonic treatment (600 W) will destroy the conformation of the protein, reducing the stability of the structure. The effect of ultrasonic treatment on emulsion gels with the ratio of SPI:HPI was <6:4 is low, but improved the gel protein digestibility. This study provides a theoretical basis for the application of ultrasonic in composite protein emulsion gels systems and the development and application of HPI.

Serum High Mobility Group Box1 Protein and Toll-like Receptor 4 Correlation with Lipid Profile in Patients with Non-alcoholic Fatty Liver Disease

Serum High Mobility Group Box1 Protein and Toll-like Receptor 4 Correlation with Lipid Profile in Patients with Non-alcoholic Fatty Liver Disease

Association of high sensitivity C-reactive protein in young myocardial patients with colchicine treatment

Association of high sensitivity C-reactive protein in young myocardial patients with colchicine treatment

Establishment of an Early Prediction Model for Severe Fever With Thrombocytopenia Syndrome-Associated Encephalitis.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease primarily transmitted by ticks. The development of encephalitis in SFTS patients significantly increases the risk of adverse outcomes. However, the understanding of SFTS-associated encephalitis (SFTSAE) is still limited. This study aimed to identify the clinical characteristics of SFTSAE and develop a predictive model for early detection. We retrospectively collected data from 220 SFTS patients admitted to Nanjing Drum Tower Hospital between May 2019 and January 2024. The patients were first randomly divided into a training set (154 people, 70%) and a validation set (66 people, 30%). The patients in the training set were divided into SFTSAE and non-SFTSAE groups according to the presence of encephalitis. A prediction model was constructed using the training set: important clinical parameters were selected using univariate logistic regression, and then multivariate logistic regression was performed to determine the independent risk factors for SFTSAE. A prediction model was constructed using these independent risk factors. Finally, the validation set was used to verify the predictive ability of the model. Age, C-reactive protein, d-dimer, and viral load were independent risk factors for SFTSAE (p < 0.05). A nomogram containing these four indicators was constructed, and the predictive performance of the nomogram was evaluated using the ROC curve. The AUC of the model was 0.846 (95% confidence interval [CI]: 0.770-0.921), which had good predictive ability for SFTSAE. Conclusion: The overall mortality rate of SFTS patients was 17.53%, and the mortality rate of encephalitis patients was 50%. Old age, high C-reactive protein, elevated d-dimer, and high viral load were independent risk factors for SFTSAE. The nomogram constructed based on these four indicators had good predictive ability and could be used as an evaluation tool for clinical treatment.

Co-treatment with erythropoietin derived HBSP and caspase-3 siRNA: A promising approach to prevent fibrosis after acute kidney injury.

Acute kidney injury (AKI) is a risk factor of chronic kidney disease, without specific treatment. This study investigated the effect of co-treatment using erythropoietin-derived helix B surface peptide (HBSP) and caspase-3 small interfering RNA (CASP3siRNA) on preventing fibrosis post AKI in order to achieve better efficacy by different action mechanisms. Ischemia-reperfusion (IR) in mice was induced by clamping bilateral renal pedicles for 30 min followed by 2-week reperfusion, with HBSP and/or CASP3siRNA administered at the onset of IR. Serum creatinine, apoptosis, active caspase-3 and high mobility group protein B1 (HMGB1) in kidneys were decreased by HBSP, CASP3siRNA or both, with increased PCNA. α-SMA expression and collagen I deposition were also reduced by CASP3siRNA and both. Most interestingly, the co-treatment further reduced tubulointerstitial damage and fibrosis, but raised PCNA compared to CASP3siRNA. EPOR/βcR was reduced by HBSP, and positively correlated with Sirius red staining, whereas EPOR was unchanged. In TCMK-1 cells, H2O2 raised apoptosis and α-SMA were reduced by HBSP, while the same was occurred to HMGB1. However, HMGB1 was further increased by EPOR siRNA under H2O2 stimulation with/without HBSP treatment. In conclusion, this study demonstrated synergistic long-term renoprotection post IR-AKI by HBSP and CASP3siRNA, which may be due to co-inhibiting inflammation and stimulating repair at early stage, and subsequently preventing fibrosis.

Efficient Development of Nanobody-Based Affinity Chromatography for AAV8 Purification

Adeno-associated virus serotype 8 (AAV8) is a highly effective vector for gene therapy. However, its purification remains challenging due to its low natural abundance and stringent purity requirements. This study aimed to develop an affinity chromatography resin utilizing nanobodies (Nbs) to enhance AAV8 purification efficiency. An AAV8-specific Nb library was constructed, leading to the identification of Nb9 as the most promising candidate based on its high binding affinity, stability and yield. Nb9 was expressed in Pichia pastoris, resulting in high yield and exceptional purity. Two types of agarose resins, Epoxy activated Bestarose 6B and PabPur SulfoLink Beads 4FF, were employed for Nb9 conjugation. Epoxy activated Bestarose 6B resin exhibited a significantly higher ligand density (9.12 mg/mL). Binding capacity assessments of the LQ01 resin demonstrated optimal performance at pH 7.0, with diminishing efficacy at lower and higher pH levels. Different NaCl concentrations influenced the binding efficiency, providing critical insights for refining purification conditions. Purification trials exhibited high specificity, purity and consistent VP protein ratio, as evidenced by SDS-PAGE analysis, confirming effective AAV8 capture and elution. Furthermore, the resin demonstrated robust performance across repeated cycles, retaining 71.9% of its initial binding capacity after 20 uses and maintaining stability with only a 6% reduction after 7 days at 37°C. These findings highlight LQ01's potential for scalable and cost-effective AAV8 purification, while demonstrating the broader applicability of Nbs in affinity chromatography and biotechnological processes.

Study on the Expression and Potential Function of LncRNA in Peripheral Blood of Patients with Ankylosing Spondylitis.

Ankylosing spondylitis (AS) is an autoimmune disease that has the characteristics of difficult early diagnosis and a high disability rate. The objective of this study was to further explore the possible mechanism and potential function of lncRNA in AS. We used lncRNA microarray technology to detect the expression of lncRNA and mRNA in patients with active AS, stable patients, and healthy controls (HC). Afterward, bioinformatics analysis was conducted on differentially expressed genes. Seven differentially expressed lncRNAs were screened out for real-time fluorescent quantitative PCR (RT-qPCR), combined with various clinical indicators for correlation analysis, and the receiver operating characteristic (ROC) curve was used to analyze the potential of lncRNA as a diagnostic marker for AS. The results showed that the expression levels of NR-037662 and ENST00000599316 in the AS subgroups were significantly higher than those in the HC group, while the expression levels of ENST00000577914 and ENST00000579003 were lower than those in the HC group. The expression levels of NR-003542 and ENST00000512051 in the ASA group were significantly higher than those in the ASS and HC groups, while NR-026756 was just the opposite. Spearman's correlation analysis showed that the expression level of NR-003542 was positively correlated with Bath Ankylosing Spondylitis Functional Index (BASFI), Erythrocyte Sedimentation Rate (ESR), and high sensitivity C-Reactive Protein (hsCRP). The expression level of NR-026756 was negatively correlated with the Bath Ankylosing Spine Inflammatory Disease Activity Index (BASDAI), BASFI, ESR, hsCRP, and globulin (GLOB). In addition, it was also found that the ROC curve analysis of the 4 lncRNAs between the AS group (ASA group and ASS group) and the HC group were statistically significant, and the area under the curve (AUC) of NR-037662, ENST00000599316, ENST00000577914, and ENST00000579003 was 0.804, 0.812, 0.706, and 0.698, respectively. It was found that these differentially expressed lncRNAs of AS may be involved in the occurrence and development of the disease. Among them, NR-037662, ENST00000599316, ENST00000577914, and ENST00000579003 might have the potential to become AS diagnostic molecular markers. Moreover, NR -003542, ENST00000512051, and NR-026756 might have the potential to be indicators of disease activity.

MDV-encoded protein kinase US3 phosphorylates WTAP to inhibit transcriptomic m6A modification and cellular protein translation

Marek’s disease virus (MDV)-encoded US3 is a highly conserved serine/threonine protein kinase in alpha-herpesviruses. In other alpha-herpesviruses, such as pseudorabies virus (PRV), US3 phosphorylates the N6-methyladenosine (m6A) methyltransferase Wilms tumor 1-associated protein (WTAP), inhibiting m6A modification. However, the role and mechanism of US3-mediated WTAP phosphorylation during MDV infection remain undefined. Our study revealed that MDV infection in vitro does not alter WTAP expression, while significant changes in WTAP expression occur during the MDV life cycle in vivo. We demonstrated that MDV-encoded US3 interacts with and co-localizes with WTAP in the nucleus. Further analysis showed that US3 binds to WTAP's C-terminal domain and phosphorylates WTAP at S273, S305, S314, and S375. Notably, the interaction between US3 and WTAP does not affect WTAP stability but inhibits transcriptomic m6A modification and cellular protein translation. Therefore, these findings enhance our understanding of the molecular mechanisms underlying MDV infection.

Comparing methods to detect cellular proteins on the surface of HIV-1 virions.

The surface of HIV-1 is embedded with numerous host-derived proteins. Characterizing these proteins can enhance knowledge of virus biology and potentially identify novel therapeutic targets. As many of these proteins are present in low abundance on virion surfaces, their identification can be hindered by inherent variables in the methods employed to detect them, including their varying assay sensitivities, sample processing, quantitative capacity, and experimental reproducibility. Here, we have compared the quantification of virion-incorporated proteins using conventional virus immunocapture assays and western blotting, alongside an emerging technique called flow virometry (FV). Using four different pseudovirus models that each express a human protein of interest (CD14, CD38, CD59 and CD162), we compared four experimental techniques for their ability to reliably quantify the incorporation of those four proteins onto virion surfaces. Our results shed light on the advantages and caveats of each technique for detecting virion-incorporated proteins and highlight the breadth in quantification for each technique under different experimental conditions. Protein detection with flow virometry provided distinct advantages as it enabled highly reproducible quantifications, had the lowest sample requirements and reagent costs, and minimal hands-on experimental time. We additionally highlight some important considerations in experimental design when studying virion-incorporated proteins, such as the effect of different antibody clones, assay incubation times, and contributions of extracellular vesicles. Most importantly, our data illustrate the importance of using a combination of orthogonal approaches to detect virus-associated proteins, to enable reliable and reproducible quantification that accounts for individual assay biases.

Characterization of the soybean oil body interface: revealing the mechanism of changes in the interfacial protein composition, structure, and function of oil bodies extracted under different denaturation conditions

We investigated the stability and rheological properties of extracts from soybean oil bodies (OB) obtained under different denaturing conditions (heating, guanidine hydrochloride, urea, and sodium dodecyl sulfate (SDS)). Differences in the interfacial and antioxidant properties exhibited by the different compositions of interfacial proteins in the extracted OB were also assessed. SDS disrupted the interfacial structure of OB, whereas the treatments under the other three denaturing conditions effectively increased the ability of the OB proteins to form a network structure. The analysis of the secondary and tertiary structures of proteins showed that all denaturation conditions affected both hydrogen bonding and hydrophobic interactions of the protein molecules, with urea leading to the most pronounced structural unfolding and rearrangement of the OB proteins. The microstructure results of the protein indicated that the protein treated with GuHCl and urea exhibited a 'shell-like' structure and structural decomposition, which can promote the pre-encapsulation of active substances through hydrogen bonding or hydrophobic interactions. The electrophoretic results further confirmed that SDS could displace 24-kDa proteins and disrupt the original membrane structure of OB, whereas urea removed most of the extrinsic proteins. The interfacial proteins extracted from OB using the four denaturing methods were different but all had enhanced interfacial and antioxidant properties compared to the control. Therefore, the extraction of OB under different denaturation conditions facilitated the hierarchical extraction of its interfacial proteins, thereby broadening its application in the food industry.

High-intensity ultrasound-assisted alkaline extraction of soy protein: Optimization, modeling, physicochemical and functional properties

This study examined the effect of high-intensity ultrasound-assisted alkaline extraction (HUAE) on the extraction yield and the physicochemical and functional properties of soy protein (SP) using the two-pot multivariate method for the first time. Plackett-Burman Design (PBD) coupled with Response Surface Methodology (RSM) was systematically utilized to select and subsequently optimize the HUAE parameters. Based on PBD results, the significant extraction factors were liquid to solid ratio (LSR), temperature, ultrasonic amplitude, and extraction time. The optimum conditions for the maximal extraction yield and minimal energy consumption were 50:1 mL/g LSR, 50 °C, 48 % ultrasonic amplitude, and 10 min extraction time. At optimum conditions, the extraction yield (35.28 %) was significantly improved compared to traditional extraction (26.39 %). Besides, HUAE resulted in modification of the protein secondary and tertiary structures due to the unfolding of protein molecules and the exposure of hydrophobic groups or regions as shown by FTIR spectroscopy, free sulfhydryl analysis, and scanning electron microscopy. These structural changes led to decreased solubility and emulsifying activity but improved emulsion stabilization and antioxidant properties. With future development, HUAE could potentially produce soy protein for targeted applications, broadening its utilization and meeting the need for more sustainable alternative processing.

Temperature dependent human serum albumin Corona formation: A case study on gold nanorods and nanospheres.

Protein molecules interact with nanoparticles to form a protein layer on the surface called the protein corona. Corona formation can be affected by the temperature and shape of the nanoparticles thereby impacting the fate of the nanoparticles inside physiological systems. We have investigated the human serum albumin (HSA) corona formation and its interactions with gold nanospheres and nanorods at different temperatures (18-42 °C). UV-Vis, fluorescence, isothermal titration calorimetry (ITC), x-ray photoelectron spectroscopy (XPS) and circular dichroism (CD) experiments have been performed to determine the changes due to the shape and temperature. UV-Vis spectra show a greater propensity of corona induced aggregation in the nanorods compared to the nanospheres. The Stern-Volmer plot indicates that static quenching is predominant in the HSA-AuNP interactions with higher quenching efficiency for nanorods than nanospheres. ITC analyses show that HSA-nanorods are involved in more favorable interactions compared to HSA-nanospheres. XPS studies indicate a more electron rich environment on the AuNRs surface and higher AuS interactions in AuNSs. CD spectra show higher secondary structural changes with temperature in case of HSA-AuNR interactions compared to HSA-AuNS interactions. The changes in the protein corona due to nanoparticle shape and variable temperature have been investigated through protein adsorption and composition, types of interactions and protein conformation.

Fabrication and characterization of acidic high internal phase emulsion gels stabilized solely by wheat gluten for plant-based mayonnaise

Plant-based diets are currently gaining more popularity as a means of reducing the environmental footprint and promoting human health. Herein, plant protein-based acidic high internal phase emulsion gels (HIPE gels) were fabricated via a facile one-pot approach using wheat gluten (WG) solely. Pre-dissolving WG in edible vinegar-water solution was contributed to the fabrication of stable emulsion gels. Factors that affect the structure and performance of plant proteins as HIPE gels stabilizers were investigated. The results indicated that the stable HIPE gels could be formed with a vinegar concentration between 5 % and 20 % and a WG concentration of 1.0 %–7.5 % only. Microscopy images reveal that a collaboration of the soluble protein molecules adsorption and protein aggregate Pickering stabilization at oil-water interface, which was turned to form stable HIPE gels. In addition, the resulted HIPE gels exhibited shear thinning behavior, viscoelastic features, and good thixotropic recovery, which provide a similar perceived texture and sensory property for formulating egg-free mayonnaise. These findings provide a useful approach to construct HIPE-based softs from low-cost wheat gluten for preparing plant-based mayonnaise.

Gradations in protein dynamics captured by experimental NMR are not well represented by AlphaFold2 models and other computational metrics.

The advent of accurate methods to predict the fold of proteins initiated by AlphaFold2 is rapidly changing our understanding of proteins and helping their design. However, these methods are mainly trained on protein structures determined with X-ray diffraction, where the protein is packed in crystals at often cryogenic temperatures. They can therefore only reliably cover well-folded parts of proteins that experience few, if any, conformational changes. Experimentally, solution nuclear magnetic resonance (NMR) is the experimental method of choice to gain insight into protein dynamics at near physiological conditions. Computationally, methods such as molecular dynamics and Normal Mode Analysis (NMA) allow the estimation of a protein's intrinsic flexibility based on a single protein structure. This work addresses, on a large scale, the relationships for proteins between the AlphaFold2 pLDDT metric, the observed dynamics in solution from NMR metrics, interpreted MD simulations, and the computed dynamics with NMA from single AlphaFold2 models and NMR ensembles. We observe that these metrics agree well for rigid residues that adopt a single well-defined conformation, which are clearly distinct from residues that exhibit dynamic behavior and adopt multiple conformations. This direct order/disorder categorisation is reflected in the correlations observed between the parameters, but becomes very limited when considering only the likely dynamic residues. The gradations of dynamics observed by NMR in flexible protein regions are therefore not represented by these computational approaches. Our results are interactively available for each protein from https://bio2byte.be/af_nmr_nma/.

Molecular displacement approach for the electrochemical detection of protein-bound propofol

Propofol is one of the principal drugs used for the sedation of patients undergoing mechanical ventilation in intensive care units. The correct dosage of such sedative drugs is highly important, but current methods of determining infusion rates are limited and there is a lack of suitable methods for directly determining patient blood propofol concentrations. A significant challenge for the development of propofol sensors is that propofol demonstrates very high protein binding, leading to a low free fraction in blood. Here we present a method for improving the efficacy of an electrochemical propofol sensor by increasing the free fraction via a molecular displacement approach. When used in conjunction with a carbon nanotube/graphene oxide/iron oxide nanoparticle functionalised screen-printed electrode, it was found that this approach dramatically improved the sensor's sensitivity towards propofol. Ibuprofen was found to be the most effective displacement agent, with an optimal concentration of 30 mM. The resultant sensitivity was 2.82 nA/μg/ml/mm2 with a coefficient of variation of 0.07, and the limit of detection was 0.2 μg/ml. This approach demonstrates high specificity towards drugs commonly administered to intensive care patients.

Camel milk whey powder formulated using thermal (spray-drying process) and non-thermal (ultrasonication) processing methods: Effect on physicochemical, technological, and functional properties

Whey protein concentrates (WPCs) are gaining importance as a functional ingredient due to their high technological and functional properties. In this study, Camel milk whey (CW) separated from skimmed camel milk, then either spray-dried (SD) at 170, 185 and 200 °C, or treated by ultrasonication (US) (20 kHz (for 5, 10 and 15 min followed by freeze-drying to obtain camel milk whey powder (CWP). The structural analysis of CWP was carried out by Fourier-Transform Infrared Spectroscopy (FTIR) and X-Ray Diffraction (XRD) which showed no significant difference in the functional groups profile of US samples compared to control and SD samples. US samples showed some degree of crystallinity that was comparable to the control samples, while SD samples exhibited very low degree of crystallinity. The surface morphology, particle size, and surface charge of CWP were evaluated using scanning electron microscopy (SEM) and Zetasizer. The lowest particle size of 215.1 nm with surface charge of −21.6 mv was observed in SD-185 WPC. Moreover, SD samples revealed whiter color compared to the US-treated samples which were having lower L* values (P < 0.05). US-15 sample exhibited high protein solubility (100 %), whereas the SD-200 sample showed reduced solubility (92.7 %). Improvement in the emulsifying activity of CWP samples was observed after SD and US, with highest emulsifying activity index (EAI) values of 143.75 m2/g and 143.11 m2/g were reported for SD-185 and US-15 CWP samples, respectively. To conclude, SD and US were found to improve the physico-chemical, technological, and functional properties of CWP, and thus can be utilized as a promising strategy to preserve and enhance techno-functional properties of CWP

Revealing mechanisms of high protein accumulation in Graesiella emersonii WBG-1 under heterotrophic condition.

Low protein content under heterotrophic conditions limits the industrial production of proteins by microalgae. In this study, Graesiella emersonii WBG-1 efficiently synthesized and accumulated proteins (64.03%) under heterotrophic conditions, distinguishing it from other microalgae. Integrated transcriptome and proteome analyses revealed that genes and proteins associated with the photosynthetic system were significantly upregulated under heterotrophic culture compared to photoautotrophic and mixotrophic conditions. Nitrogen assimilation was enhanced while carbohydrate and fatty acid biosynthesis were restricted, carbon redirected towards amino acid and protein synthesis. Ribosome biogenesis was strengthened, and translation initiation and elongation factors were upregulated, increasing the translational activity of algal cells and promoting overall protein synthesis. Overall, these findings elucidate the mechanisms underlying efficient protein synthesis in G. emersonii WBG-1 under heterotrophic conditions, offering new insights and complementary perspectives on the regulation of protein synthesis in microalgae across different nutritional modes.