The Effect of pH and Sodium Caseinate on the Aqueous Solubility,Stability, and Crystallinity of Rutin towards Concentrated Colloidally Stable Particles for the Incorporation into Functional Foods

Poor water solubility and low bioavailability of hydrophobic flavonoids such as rutin remain as substantial challenges to their oral delivery via functional foods. In this study, the effect of pH and the addition of a protein (sodium caseinate; NaCas) on the aqueous solubility and stability of rutin was studied, from which an efficient delivery system for the incorporation of rutin into functional food products was developed. The aqueous solubility, chemical stability, crystallinity, and morphology of rutin (0.1–5% w/v) under various pH (1–11) and protein concentrations (0.2–8% w/v) were studied. To manufacture the concentrated colloidally stable rutin–NaCas particles, rutin was dissolved and deprotonated in a NaCas solution at alkaline pH before its subsequent neutralisation at pH 7. The excess water was removed using ultrafiltration to improve the loading capacity. Rutin showed the highest solubility at pH 11, while the addition of NaCas resulted in the improvement of both solubility and chemical stability. Critically, to achieve particles with colloidal stability, the NaCas:rutin ratio (w/w) had to be greater than 2.5 and 40 respectively for the lowest (0.2% w/v) and highest (4 to 8% w/v) concentrations of NaCas. The rutin–NaCas particles in the concentrated formulations were physically stable, with a size in the range of 185 to 230 nm and zeta potential of −36.8 to −38.1 mV, depending on the NaCas:rutin ratio. Encapsulation efficiency and loading capacity of rutin in different systems were 76% to 83% and 2% to 22%, respectively. The concentrated formulation containing 5% w/v NaCas and 2% w/v rutin was chosen as the most efficient delivery system due to the ideal protein:flavonoid ratio (2.5:1), which resulted in the highest loading capacity (22%). Taken together, the findings show that the delivery system developed in this study can be a promising method for the incorporation of a high concentration of hydrophobic flavonoids such as rutin into functional foods.     

Caseinolytic Proteins (Clp) in the Genus Klebsiella: Special Focus on ClpK

Caseinolytic proteins (Clp), which are present in both prokaryotes and eukaryotes, play a major role in cell protein quality control and survival of bacteria in harsh environmental conditions. Recently, a member of this protein family, ClpK was identified in a pathogenic strain of Klebsiella pneumoniae which was responsible for nosocomial infections. ClpK is linked to the thermal stress survival of this pathogen. The genome wide analysis of Clp proteins in Klebsiella spp. indicates that ClpK is present in only 34% of the investigated strains. This suggests that the uptake of the clpk gene is selective and may only be taken up by a pathogen that needs to survive harsh environmental conditions. In silico analyses and molecular dynamic simulations show that ClpK is mainly α-helical and is highly dynamic. ClpK was successfully expressed and purified to homogeneity using affinity and anion exchange chromatography. Biophysical characterization of ClpK showed that it is predominantly alpha-helical, and this is in agreement with in silico analysis of the protein structure. Furthermore, the purified protein is biologically active and hydrolyses ATP in a concentration- dependent manner.     

Natural Polymers-Based Materials: A Contribution to a Greener Future

Natural polymers have emerged as promising candidates for the sustainable development of materials in areas ranging from food packaging and biomedicine to energy storage and electronics. In tandem, there is a growing interest in the design of advanced materials devised from naturally abundant and renewable feedstocks, in alignment with the principles of Green Chemistry and the 2030 Agenda for Sustainable Development. This review aims to highlight some examples of the research efforts conducted at the Research Team BioPol4fun, Innovation in BioPolymer-based Functional Materials and Bioactive Compounds, from the Portuguese Associate Laboratory CICECO–Aveiro Institute of Materials at the University of Aveiro, regarding the exploitation of natural polymers (and derivatives thereof) for the development of distinct sustainable biobased materials. In particular, focus will be given to the use of polysaccharides (cellulose, chitosan, pullulan, hyaluronic acid, fucoidan, alginate, and agar) and proteins (lysozyme and gelatin) for the assembly of composites, coatings, films, membranes, patches, nanosystems, and microneedles using environmentally friendly strategies, and to address their main domains of application.     

淀粉样蛋白聚集纤维化的抑制作用研究

总结了不同抑制剂对淀粉样蛋白聚集及纤维化的抑制作用,主要介绍了金属配合物作为淀粉样蛋白抑制剂的研究,并概述了淀粉样蛋白相互作用体系的热力学研究进展.     

Accounting for the energies and entropies of kinesin’s catalytic cycle

When the processive motor protein kinesin walks along the biopolymer microtubule it can occasionally make a backward step. Recent single molecule experiments on moving kinesin have revealed that the forward-to-backward step ratio decreases exponentially with the load force. Carter and Cross (Nature 435, 308-312, 2005) found that this ratio tightly followed 802 × exp[−0.95F], where F is the load force in piconewtons. A straightforward analysis of a Brownian step leads to L/(2k _B T) as the factor in front of the load force, where L is the 8 nm stepsize, k _B is the Boltzmann constant, and T is the temperature. The factor L/(2k _B T) does indeed equal 0.95 pN⁻¹. The same analysis shows how the 802 prefactor derives from the power stroke energy G as exp[G/(2k _B T)]. There are indications that the power stroke derives from the entropically driven coiling of the 30 amino acid neck linker that connects the two kinesin heads. This idea is examined and consequences are deduced.     

Proteomic response and molecular regulatory mechanisms of Bacillus cereus spores under ultrasound treatment

This study was aimed at providing new insights on the proteomic response of bacterial spores to ultrasound. Data-independent-acquisition method was used to quantify the proteome change of Bacillus cereus spores after ultrasound treatment (200 W). This study revealed that 2485 proteins were extracted from Bacillus cereus spores, most of them were related to metabolism. After ultrasound treatment, the expression of 340 proteins were significantly changed (the fold change ≥ 2 and p < 0.05), of which 207 proteins were significantly down-regulated. KEGG pathway analysis showed that differentially expressed proteins mainly distributed in metabolism pathway, cell process pathway and genetic information processing pathway after ultrasound treatment. Furthermore, this study analyzed the differentially expressed proteins in significant enrichment pathways. In particular, the expression of key proteins in the phosphorylation reaction of spores was significantly decreased after ultrasound treatment. Thus, ATP synthesis rate decreased and the phosphorylation reaction inhibited. Also, the decrease of the expression of key proteins related to the tricarboxylic acid cycle led to the decrease of nutrients metabolism of spores. Ultrasound treatment induced the down-regulation of fatty acid synthetase expression and promoted fatty acid metabolism at the same time. The content of fatty acids decreased in spores consequently.     

Theorie und Optimierung der radiometrischen Absorptionsmethode zur Analyse von Binärsystemen

Die Arbeit behandelt theoretisch das Auflösungsvermögen der Absorptionsmethode zur Analyse von Binārsystemen bei Anwendung von monoenergetischer Gamma- und Rötgenstrahlung. Es wird eine analytische Beziehung für die Funktion Δp abgeleitet, die die kleinstmöglichen Veränderung in der Zusammensetzung des Systems chgarakterisiert und die wir noch mittelts dieser Methode in Abhängigkeit von allen zu berückischtigenden Faktoren bestimmen könne. Aus den Extremwerten der Funktion Δp werden dann die optimalen Werte der Energine, der Schichtdicke, der Strahlungshäufigkeit und die Optimalwerte der restlichen Parameter so bestimmt, daβ das Auflösungsvermögen der Methode maximal ist. Die Theorie wird auf eine Ag-Cu-Legierung angewendet.     

Trichoderma reesei cellulases and their core domains in the hydrolysis and modification of chemical pulp

The action of monocomponent Trichoderma reesei endoglucanases (EG I, EG II; EC 3.2.1.4) and cellobiohydrolases (CBH I, CBH II; EC 3.2.1.91) and their core proteins was compared using isolated celluloses and bleached chemical pulp. The presence of cellulose binding domain (CBD) in the intact enzymes did not affect their action against soluble substrates. In the case of insoluble isolated celluloses and the chemical pulp the presence of CBD enhanced the enzymatic hydrolysis of cellulose. The effect of CBD was more pronounced in the cellobiohydrolases, hydrolysing mainly crystalline cellulose, than in the endoglucanases which were more efficient in hydrolysing amorphous cellulose. The pulp properties measured, that is, viscosity and strength after PFI refining, were equally affected by the treatment with intact enzymes and corresponding core proteins, suggesting that the presence of CBD in intact cellulases affects mainly the cellulose hydrolysis level and less the mode of action of T. reesei cellulases in pulp. The better beatability of the bleached chemical pulp treated with intact endoglucanases than that treated with the corresponding core proteins suggests that the presence of CBD in endoglucanases could, however, result in beneficial effects on pulp properties.