E. coli relies on the heat shock response (HSR) to preserve protein homeostasis under stress, through three feedback modules: feedforward translational control, chaperone-mediated sequestration and targeted degradation. Although previous studies have highlighted how this layered architecture ensures rapid and robust protection compared to simpler designs, not much attention is paid to how these modules interact. Moreover, how do interactions among the three modules balance performance trade-offs, where gains in one module may come at the expense of another, yet together yield an optimal overall response? We address this using a mathematical model that integrates protein folding with σ 32 regulation. We show that the feedback modules both cooperate and compete, giving rise to nonmonotonic dynamics that govern HSR performance. Specifically, increasing feedforward strength does accelerate response, but beyond a threshold, despite increasing chaperone levels, it paradoxically slows recovery. Similarly, while sequestration enhances relative chaperone production and per-chaperone efficiency, when excessive, it traps σ 32 in inactive complexes, prolonging recovery and delaying shutdown. Mapping the parameter space reveals regimes of synergy as well as trade-offs between speed and efficiency, with wild-type parameters lying near the optimal region. These results reveal design principles that produces a robust and efficient heat shock response.
Uremic conditions are common in end-stage kidney disease (ESKD) patients. Accelerated vascular diseases in uremic patients lead to heart failure, stroke, and hypertension. To investigate the effects of uremia on porcine arterial smooth muscle cells (aSMCs), bulk RNA sequencing was used to identify uremia-induced alterations in signaling pathways of aSMCs that might explain the aggressive cardiovascular diseases seen in patients with chronic kidney disease (CKD) and ESKD. Bulk RNA sequencing was performed on porcine aSMCs cultured with serum from normal or uremic pigs. Differentially expressed gene (DEG) analysis revealed that 295 genes were upregulated and 138 genes were downregulated after uremic serum exposure. Gene Ontology molecular function analysis demonstrated that ATP-dependent activity, translation factor activity, and ATP-dependent protein folding chaperones were predicted to be negatively enriched after uremic serum exposure, while proton transmembrane transporter activity, antioxidant activity, and glutathione peroxidase activity were predicted to be positively enriched. Gene set enrichment analysis indicated that the cell cycle was predicted to be negatively enriched after uremic serum exposure in aSMCs. Overrepresentation analysis found that focal adhesion, protein processing in the endoplasmic reticulum (ER) and cell senescence were predicted to be negatively enriched, while lysosome, phagosome, apoptosis, and autophagy were predicted to be positively enriched after uremic serum exposure. This study suggests that the signaling pathways that regulate cellular redox homeostasis, the cellular waste disposal system, ER stress and autophagy are major signaling pathways involved in aSMCs’ responses to uremic serum exposure. These pathways may contribute to the severe arterial-specific clinical symptoms observed in CKD/ESKD patients, such as arterial stiffness, vascular calcification and cardiovascular disease.
Unimunkh Uriyanghai, Huanjuan Su, John S. Poulton et al.· UNC Libraries· 0 citations
Temporal Wave Function Collapse Dynamics explores the theoretical underpinnings of the collapse of temporal wave functions – fundamental units of information within complex systems such as neural networks and protein folding – as a dynamic process. This paper posits that collapse isn't a discrete event but rather a continuous evolution driven by a set of differential equations that capture the interplay between system state, external stimuli, and feedback loops. We propose a novel differential equation system that models this collapse, emphasizing the generation of new, potentially transformative states. This research aims to advance our understanding of complex system behavior by providing a framework for modeling this fundamental process.
Jincheng Zhang· Zenodo (CERN European Organi...· 0 citations
Molecular dynamics (MD) simulations are a fundamental tool in materials science, biology, and pharmaceutical research, offering insights into molecular behavior and dynamics. However, traditional MD simulations often suffer from limitations in accuracy and efficiency, particularly when dealing with complex, dynamic environments. This paper introduces a novel algorithm, termed Adaptive Molecular Dynamics Optimization (AMDO), designed to address these challenges by dynamically adjusting simulation parameters based on a learned model. AMDO leverages an adaptive learning algorithm to optimize the simulation process, resulting in enhanced accuracy and reduced computational cost. We demonstrate the effectiveness of AMDO through the simulation of a complex protein folding process, showcasing improved convergence and reduced simulation time compared to conventional MD methods. The core mechanism centers on the continuous adaptation of simulation parameters, enabling the simulation to effectively capture the nuances of molecular interactions.
Jincheng Zhang· Zenodo (CERN European Organi...· 0 citations
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Colorectal cancer (CRC) is a widespread health issue that attains high mortality. The adaptor protein SH3BP2 amplification results in metabolic changes, oxidative stress, NK cell activity, and inflammation. The NK cells are capable of destroying tumor cells without prior activation, help prevent metastasis, and have prognostic value. Targeting SH3BP2 to regulate NK cell activity in the TME could enhance CRC-based immunotherapy. The cancer hallmark tool helps in understanding SH3BP2 hallmark annotation. Utilizing the STRING tool and the KEGG pathway, protein functional enrichment and PPI networking were analyzed. TIMER 2.0 was used for immune cell infiltration correlation analysis, and UALCAN was used for CPTAC-based protein expression profiling. The GEO (GSE9348) dataset showed SH3BP2 is upregulated in CRC (log2 fold change = 1.18). GEO, TCGA, and cBioPortal revealed SH3BP2 alterations in CRC cases, potentially aiding immune evasion. Mutations in SH3BP2 influence cancer growth, suppressing tumors or promoting them by activating NF-κB and affecting immune responses through WNT/β-catenin, PI3K, MAPK, and JAK-STAT pathways. Overall, SH3BP2 plays a key role in cancer growth and immune regulation, making it a promising target for CRC therapy. Further experimental validation is needed to demonstrate its diagnostic and therapeutic potency.
Researchers at Zhejiang University in China and Imperial College London in the UK built standalone tunnelling electrodes with an average gap of 1.6 nm at the tip of a nanopipette, and used them to identify single nucleotides and proteins electrically. By combining the probes with dielectrophoretic (DEP) trapping, which uses an alternating electric field to concentrate molecules, the team got past the diffusion limit, raising event detection rates by up to five orders of magnitude (100,000-fold) and reaching sub-femtomolar (fM) sensitivity. The paper frames nucleic acid sequencing as a future prospect rather than something achieved here, and notes that at the lowest concentrations the same molecules are likely being recaptured near the probe tip and detected repeatedly. [Quantum Biology Society] Quantum tunnelling, in which an electron passes through a potential barrier it could not classically cross by virtue of its wave nature, has long served as a high-precision measurement tool, because the current varies exponentially with the width of the gap and is therefore sensitive to changes in distance at the atomic scale. Forming a gap narrower than 5 nm between nanoelectrodes and reading the characteristic tunnelling current of a single molecule passing through it has raised hopes for a next generation of nucleic acid sequencing and, potentially, even protein sequencing. Existing approaches based on the scanning tunnelling microscope (STM), however, require a conductive substrate and precise piezo controllers, which makes the system unwieldy. They also have to wait for molecules to diffuse into a gap only nanometres wide, a process that is entropically unfavourable, so the efficiency of real sample analysis has been extremely low. A collaboration led by Longhua Tang at Zhejiang University in China, together with Aleksandar P. Ivanov and Joshua B. Edel at Imperial College London, published a standalone nanoprobe platform in Nature Communications in February 2021 that addresses both limitations at once. ■ Self-Terminating Electrodeposition: Forming Nanometre Gaps With Precision The researchers laser-pulled a theta-shaped dual-barrel quartz capillary into a nanopipette whose tip terminates in two closely spaced nanopores, 25 ± 12 nm in diameter, separated by a quartz septum 15 ± 5 nm wide. Butane was then passed through the pipette and pyrolytically deposited to form two coplanar carbon nanoelectrodes, onto which gold was electrochemically plated. The key is a self-terminating mechanism that applies tunnelling current feedback during plating. The preset current is the sum of a Faradaic deposition component and a tunnelling component. As the gap between the electrodes narrows into the tunnelling regime, the tunnelling component comes to dominate, the Faradaic current falls towards zero, and deposition stops of its own accord. Of the 650 probes fabricated, roughly 85% ran through self-termination successfully. The freshly made gaps were then immersed in ultrapure deionised water for 12 to 48 hours, after which conductance had dropped by an average of 55%, consistent with a widening of the tunnelling gaps. The authors tentatively attribute this change to surface diffusion of gold atoms minimising the total interfacial free energy at a fixed volume. The resulting probes held consistent I-V characteristics over several days. Fitted with the Simmons model, 418 standalone probes had gap widths ranging from sub-nanometre to more than 3 nm, with an average of 1.6 ± 0.6 nm. ■ Verifying Tunnelling by Measuring Solvent Barrier Heights To check that the current flowing across the fabricated gaps really came from tunnelling, the researchers measured the tunnelling potential barrier of the surrounding medium. The values came out at 0.37 ± 0.21 eV for deionised water, 0.78 ± 0.14 eV for dimethyl sulfoxide and 0.97 ± 0.21 eV for hexane, all in agreement with the literature. In air the figure was 1.04 ± 0.83 eV, and the large spread was attributed to possible condensation of water vapour in the gap. In control experiments with bridged, short-circuited junctions, the I-V characteristics showed no dependence on the medium, confirming that the junctions were working as tunnelling junctions rather than through physical contact between the electrodes. ■ Identifying Nucleotides and Proteins by Their Characteristic Conductance Measuring four deoxymononucleotides with a probe of roughly 1.1 nm gap at a bias of 50 mV, the team found a clear ordering in the conductance change (ΔG): dGMP (240 ± 36 nS) > dAMP (180 ± 33 nS) > dCMP (161 ± 5 nS) > dTMP (120 ± 10 nS). The authors offer a partial, qualitative interpretation in terms of the highest occupied molecular orbital (HOMO): dGMP shows the highest conductance because its HOMO level sits closer to the Fermi level of the electrodes. They are careful to add that the actual electron transport mechanism across mononucleotides remains an open question. In a mixed solution of dTMP and dGMP, the two characteristic conductance peaks separated clearly. With a probe of roughly 1.8 nm gap, three proteins chosen for their differing molecular weights and charges were likewise told apart by clearly different conductance values: streptavidin (1.56 ± 0.19 nS), bovine serum albumin (BSA, 1.11 ± 0.21 nS) and immunoglobulin G (IgG, 0.52 ± 0.08 nS). Although the gap was narrower than the proteins themselves, characteristic tunnelling signals were still obtained. ■ Combining Dielectrophoresis: Molecular Trapping and a Five-Order Gain in Detection Rate The biggest obstacle to single-molecule sensing in a nanogap, the mass transport limit, was tackled with dielectrophoresis (DEP). Applying an AC field between the two electrodes (100 kHz, 10 Vpp, 10 seconds) generates exceptionally steep field gradients that pull target molecules in solution towards the probe tip and concentrate them there. Measurements could not be run in parallel, because applying the AC field brought a significant rise in the low-frequency noise associated with conductance fluctuations (flicker noise), along with a milder increase in higher-frequency noise attributed to capacitance. The team therefore ran DEP concentration and DC tunnelling detection sequentially. At a poly-A20 DNA concentration of 1 fM, no significant tunnelling events were seen without DEP, whereas clear spike signals appeared after trapping, and event detection rates rose by up to five orders of magnitude. That brought detection down to 0.1 fM for poly-A20 and 0.15 fM for streptavidin. ■ Significance and Limits: Single-Molecule Identification and the Recapture Mechanism The significance of the work lies in realising an ultrasensitive nanoscale tunnelling sensor that operates in solution as a standalone capillary probe, breaking away from the conventional STM architecture in which a conductive substrate is essential. The authors also state that, to their knowledge, this is the first example of dynamic control of molecular transport used in any tunnelling system. The paper treats nucleic acid sequencing as a future prospect rather than something completed here; what is demonstrated is the identification of single mononucleotides and the detection of oligomers and proteins. The authors also note that at the lowest concentrations probed, around 0.1 fM, the event rate is very high, above 200 events per second, and loses any significant dependence on concentration. They explain this as likely arising because trapped molecules localise and accumulate around the tip, so the same molecules have a much higher probability of being recaptured and interacting with the gap, and are probably detected multiple times. Read that way, in this regime the platform looks less like a quantitative counting instrument and more like an ultrasensitive qualitative test for the presence of trace molecules. #QuantumTunnelling #Dielectrophoresis #SingleMoleculeDetection #Nanoelectrode #TunnellingCurrent #Nucleotide #ProteinConductance #SimmonsModel #Nanopipette #BiomolecularSensing #QuantumBiology #NatureCommunications Source (Nature Communications): https://www.nature.com/articles/s41467-021-21101-x Follow-up study (Sci. Adv. 2022): https://doi.org/10.1126/sciadv.abm8149
inquantio· Zenodo (CERN European Organi...· 0 citations
Researchers at Zhejiang University in China and Imperial College London in the UK built standalone tunnelling electrodes with an average gap of 1.6 nm at the tip of a nanopipette, and used them to identify single nucleotides and proteins electrically. By combining the probes with dielectrophoretic (DEP) trapping, which uses an alternating electric field to concentrate molecules, the team got past the diffusion limit, raising event detection rates by up to five orders of magnitude (100,000-fold) and reaching sub-femtomolar (fM) sensitivity. The paper frames nucleic acid sequencing as a future prospect rather than something achieved here, and notes that at the lowest concentrations the same molecules are likely being recaptured near the probe tip and detected repeatedly. [Quantum Biology Society] Quantum tunnelling, in which an electron passes through a potential barrier it could not classically cross by virtue of its wave nature, has long served as a high-precision measurement tool, because the current varies exponentially with the width of the gap and is therefore sensitive to changes in distance at the atomic scale. Forming a gap narrower than 5 nm between nanoelectrodes and reading the characteristic tunnelling current of a single molecule passing through it has raised hopes for a next generation of nucleic acid sequencing and, potentially, even protein sequencing. Existing approaches based on the scanning tunnelling microscope (STM), however, require a conductive substrate and precise piezo controllers, which makes the system unwieldy. They also have to wait for molecules to diffuse into a gap only nanometres wide, a process that is entropically unfavourable, so the efficiency of real sample analysis has been extremely low. A collaboration led by Longhua Tang at Zhejiang University in China, together with Aleksandar P. Ivanov and Joshua B. Edel at Imperial College London, published a standalone nanoprobe platform in Nature Communications in February 2021 that addresses both limitations at once. ■ Self-Terminating Electrodeposition: Forming Nanometre Gaps With Precision The researchers laser-pulled a theta-shaped dual-barrel quartz capillary into a nanopipette whose tip terminates in two closely spaced nanopores, 25 ± 12 nm in diameter, separated by a quartz septum 15 ± 5 nm wide. Butane was then passed through the pipette and pyrolytically deposited to form two coplanar carbon nanoelectrodes, onto which gold was electrochemically plated. The key is a self-terminating mechanism that applies tunnelling current feedback during plating. The preset current is the sum of a Faradaic deposition component and a tunnelling component. As the gap between the electrodes narrows into the tunnelling regime, the tunnelling component comes to dominate, the Faradaic current falls towards zero, and deposition stops of its own accord. Of the 650 probes fabricated, roughly 85% ran through self-termination successfully. The freshly made gaps were then immersed in ultrapure deionised water for 12 to 48 hours, after which conductance had dropped by an average of 55%, consistent with a widening of the tunnelling gaps. The authors tentatively attribute this change to surface diffusion of gold atoms minimising the total interfacial free energy at a fixed volume. The resulting probes held consistent I-V characteristics over several days. Fitted with the Simmons model, 418 standalone probes had gap widths ranging from sub-nanometre to more than 3 nm, with an average of 1.6 ± 0.6 nm. ■ Verifying Tunnelling by Measuring Solvent Barrier Heights To check that the current flowing across the fabricated gaps really came from tunnelling, the researchers measured the tunnelling potential barrier of the surrounding medium. The values came out at 0.37 ± 0.21 eV for deionised water, 0.78 ± 0.14 eV for dimethyl sulfoxide and 0.97 ± 0.21 eV for hexane, all in agreement with the literature. In air the figure was 1.04 ± 0.83 eV, and the large spread was attributed to possible condensation of water vapour in the gap. In control experiments with bridged, short-circuited junctions, the I-V characteristics showed no dependence on the medium, confirming that the junctions were working as tunnelling junctions rather than through physical contact between the electrodes. ■ Identifying Nucleotides and Proteins by Their Characteristic Conductance Measuring four deoxymononucleotides with a probe of roughly 1.1 nm gap at a bias of 50 mV, the team found a clear ordering in the conductance change (ΔG): dGMP (240 ± 36 nS) > dAMP (180 ± 33 nS) > dCMP (161 ± 5 nS) > dTMP (120 ± 10 nS). The authors offer a partial, qualitative interpretation in terms of the highest occupied molecular orbital (HOMO): dGMP shows the highest conductance because its HOMO level sits closer to the Fermi level of the electrodes. They are careful to add that the actual electron transport mechanism across mononucleotides remains an open question. In a mixed solution of dTMP and dGMP, the two characteristic conductance peaks separated clearly. With a probe of roughly 1.8 nm gap, three proteins chosen for their differing molecular weights and charges were likewise told apart by clearly different conductance values: streptavidin (1.56 ± 0.19 nS), bovine serum albumin (BSA, 1.11 ± 0.21 nS) and immunoglobulin G (IgG, 0.52 ± 0.08 nS). Although the gap was narrower than the proteins themselves, characteristic tunnelling signals were still obtained. ■ Combining Dielectrophoresis: Molecular Trapping and a Five-Order Gain in Detection Rate The biggest obstacle to single-molecule sensing in a nanogap, the mass transport limit, was tackled with dielectrophoresis (DEP). Applying an AC field between the two electrodes (100 kHz, 10 Vpp, 10 seconds) generates exceptionally steep field gradients that pull target molecules in solution towards the probe tip and concentrate them there. Measurements could not be run in parallel, because applying the AC field brought a significant rise in the low-frequency noise associated with conductance fluctuations (flicker noise), along with a milder increase in higher-frequency noise attributed to capacitance. The team therefore ran DEP concentration and DC tunnelling detection sequentially. At a poly-A20 DNA concentration of 1 fM, no significant tunnelling events were seen without DEP, whereas clear spike signals appeared after trapping, and event detection rates rose by up to five orders of magnitude. That brought detection down to 0.1 fM for poly-A20 and 0.15 fM for streptavidin. ■ Significance and Limits: Single-Molecule Identification and the Recapture Mechanism The significance of the work lies in realising an ultrasensitive nanoscale tunnelling sensor that operates in solution as a standalone capillary probe, breaking away from the conventional STM architecture in which a conductive substrate is essential. The authors also state that, to their knowledge, this is the first example of dynamic control of molecular transport used in any tunnelling system. The paper treats nucleic acid sequencing as a future prospect rather than something completed here; what is demonstrated is the identification of single mononucleotides and the detection of oligomers and proteins. The authors also note that at the lowest concentrations probed, around 0.1 fM, the event rate is very high, above 200 events per second, and loses any significant dependence on concentration. They explain this as likely arising because trapped molecules localise and accumulate around the tip, so the same molecules have a much higher probability of being recaptured and interacting with the gap, and are probably detected multiple times. Read that way, in this regime the platform looks less like a quantitative counting instrument and more like an ultrasensitive qualitative test for the presence of trace molecules. #QuantumTunnelling #Dielectrophoresis #SingleMoleculeDetection #Nanoelectrode #TunnellingCurrent #Nucleotide #ProteinConductance #SimmonsModel #Nanopipette #BiomolecularSensing #QuantumBiology #NatureCommunications Source (Nature Communications): https://www.nature.com/articles/s41467-021-21101-x Follow-up study (Sci. Adv. 2022): https://doi.org/10.1126/sciadv.abm8149
inquantio· Zenodo (CERN European Organi...· 0 citations
Derived data, run outputs and figure source data supporting the article "Joint optimisation of amino acid and coding sequence for de novo designed proteins". Includes the inverse-folding marginals for all 862 backbones and the full 230,992-row double-mutant additivity table.
Anees Ahmed Mahaboob Ali, Radhakrishnan Delhibabu, Everette Jacob Remington Nelson· Zenodo (CERN European Organi...· 0 citations
Alfalfa biomass contains significant carbohydrate fractions underutilized in animal feed. This research optimized the enzymatic hydrolysis of Alfalfa biomass and evaluated single-cell protein (SCP) production using Candida utilis and Komagataella pastoris . Hydrolysis kinetics in this study showed biphasic sugar release, with enzymatic optimization increasing monomeric sugar yield from 18.5% to a maximum of 33.0% (a 78% relative increase) at 22 mg protein/g biomass enzyme loading. C. utilis demonstrated superior metabolic versatility, consuming 23% more total sugars than K. pastoris , with particularly enhanced pentose utilization showing 45% greater xylose consumption. K. pastoris achieved 51% faster growth rates than C. utilis , while both yeasts produced comparable protein yields per unit sugar consumed. SCP production enriched protein content 2.1-fold compared to raw Alfalfa biomass, reaching approximately 40% crude protein. Essential amino acid profiling (tryptophan excluded) showed that fermentation substantially improved several essential amino acids relative to FAO/WHO reference values, particularly lysine, threonine, and methionine; however, methionine + cysteine remained below the FAO reference ratio in both yeasts ( K. pastoris : 0.99; C. utilis : 0.87), indicating that sulfur-containing amino acids remain a limiting factor despite overall nutritional improvement. The integrated bioprocess achieved 27.6% carbohydrate-to-biomass conversion efficiency and 11.3% carbohydrate-to-protein conversion efficiency based on measured monomeric sugars (oligosaccharide utilization during fermentation was not independently quantified). This work demonstrates a promising lab-scale strategy for alfalfa valorization through enzymatic hydrolysis and yeast fermentation, yielding a nutritionally improved protein product; techno-economic analysis, feeding trials, and scale-up studies are required before cost-effectiveness or industrial readiness can be established.
Shehnaz kousar, Saddam Hussain, Qurban Ali et al.· AMB Express· 0 citations
Glucoamylases are highly demanded by the industry for starch saccharification, which necessitates an increase in enzyme activity. To improve the activity of Aspergillus awamori VKPM F-1262 glucoamylase N181Q (AGAM), amino acid substitutions were made in the region of substrate binding. In the case of the variant D237G, there was a 1.3–1.4-fold increase in activity towards soluble starch for the individual enzyme and crude enzyme solution after cultivation in flasks. For the variants G57A and C319T, there was also a 1.1–1.4-fold increase in enzyme activity for the crude flask solution, while the increase in individual enzyme activity was minor. AGAM showed an increase in apparent molecular weight consistent with dimerization in the presence of soluble starch. A high T optimum of 70 °C was observed for AGAM and the variants in the hydrolysis of soluble starch. In further protein engineering, a high T optimum for hydrolysis, which may coincide with dimerization, may be considered as a hypothesis for the selection of thermostable glucoamylases, while an area of binding of oligo- and polysaccharides may be a site for mutagenesis.
Anna S. Dotsenko, Nikita Eroshenko, Ekaterina Rubtsova et al.· BioTech· 0 citations
Eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein family unique to eukaryotes, yet its functional characterization in woody plants remains limited. In this study, we identified four eIF5A genes (PtoeIF5A1–PtoeIF5A4) from the genome of Populus tomentosa, a fast-growing tree species indigenous to China, and characterized their expression patterns and functional roles through bioinformatics analysis, quantitative real-time PCR, stable overexpression in Arabidopsis thaliana, and transient expression in Nicotiana benthamiana leaves. Our results demonstrated that all PtoeIF5A proteins contain a conserved OB-fold domain and multiple phosphorylation sites, with PtoeIF5A1 showing predominant expression in roots and secondary xylem. Functional assays revealed that PtoeIF5A1 overexpression accelerated inflorescence stem elongation and early flowering in Arabidopsis, induced visible chlorosis and programmed cell death (PCD) in tobacco leaves, and significantly enhanced salt tolerance under NaCl treatment. Collectively, these findings establish PtoeIF5A1 in poplar as a pleiotropic regulator integrating developmental cues, programmed cell death, and stress responses; and as a valuable genetic resource for breeding stress-resilient woody plants.
Dan Zhu, Guan Yang, Feng Feng et al.· Plants· 0 citations
Abstract Probiotic efficacy depends not only on gastrointestinal survival but on mucosal adhesion and the capacity to deliver bioactive molecules at the intestinal surface. This study optimized a whey protein isolate (WPI)–chitosan (CS) matrix for spray drying microencapsulation of Lacticaseibacillus casei BL23 using central composite design, targeting enhanced mucoadhesion while preserving bacterial viability and extracellular vesicle (EV) secretion capacity. The optimal formulation, WPI 20%-CS 0.5%, yielded viable counts within recommended probiotic ranges (6.6 × 10⁹ CFU/g) and a ~ 77-fold mucoadhesion increase relative to WPI alone, supporting extended intestinal residence time and potentially enhanced therapeutic efficacy. Storage stability was confirmed at 4°C and − 20°C, and microcapsules were predominantly spherical (2–15 μm), suitable for food applications. Microencapsulation also significantly enhanced gastrointestinal survival, with encapsulated bacteria showing only a ~ 2-log reduction after the gastric phase compared to ~ 6-log for free bacteria. Fermentative capacity in reconstituted milk was fully preserved, with reduced syneresis indicative of improved gel stability. Critically, this system ensures the delivery of probiotic-derived extracellular vesicles (EVs) at the intestinal interface. We demonstrate that EV secretion is maintained post-encapsulation, yielding vesicles (70–100 nm) enriched in p40 and p75 proteins. To our knowledge, this is the first evidence that spray-dried microcapsules can effectively serve as a delivery platform for postbiotic EVs by preserving the functional secretory machinery of the encapsulated bacteria. These findings position mucoadhesive WPI-CS microcapsules as a robust strategy for the targeted delivery of EVs in nutraceutical and functional food development. Key points • Whey protein-chitosan microencapsulation boosts mucoadhesion ~77-fold in vitro. • Microencapsulation enhances probiotic survival across gastrointestinal conditions. • Spray-drying preserves the probiotic machinery for postbiotic EV secretion.
Cecilia L. D’Antoni, Rocío Corfield, Sergio I. Nemirovsky et al.· Applied Microbiology and Bio...· 0 citations
A new machine-learning framework aims to improve the success rate of computational protein design while moving away from results that reproduce sequences found in nature.
