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Amyloid-β (Aβ) peptide and tau protein are two hallmark proteins in Alzheimer's disease (AD); however, the parameters, which mediate the abnormal aggregation of Aβ and tau, have not been fully discovered. Here, we have provided an optimum method to purify tau protein isoform 1N4R by using nickel-nitrilotriacetic acid agarose chromatography under denaturing condition. The biochemical and biophysical properties of the purified protein were further characterized using in vitro tau filament assembly, tubulin polymerization assay, circular dichroism (CD) spectroscopy and atomic force microscopy. Afterwards, we investigated the effect of tau protein on aggregation of Aβ (25–35) peptide using microscopic imaging and cell viability assay. Incubation of tau at physiologic and supra-physiologic concentrations with Aβ25–35 for 40 days under reducing and non-reducing conditions revealed formation of two types of aggregates with distinct morphologies and dimensions. In non-reducing condition, the co-incubated sample showed granular aggregates, while in reducing condition, they formed annular protofibrils. Results from cell viability assay revealed the increased cell viability for the co-incubated sample. Therefore, the disassembling action shown by tau protein on Aβ25–35 suggests the possibility that tau may have a protective role in preventing Aβ peptide from acquiring the cytotoxic, aggregated form against oxidative stress damages. [ABSTRACT FROM AUTHOR]

The effects of 17 kinds of additive mixtures have been studied on refolding and aggregation of a model protein, lysozyme. Most of the prepared mixtures were efficient in inhibiting aggregation of the protein, and, surprisingly, four novel additive mixtures, i.e., lactic acid: l‐arginine, lactic acid: l‐glutamine, choline chloride: lactic acid, and imidazolium salt: β‐cyclodextrin as well as choline chloride: urea exhibited a more remarkable efficacy in suppressing aggregation. Among these, lactic acid: l‐arginine was identified as the most efficient additive, and lactic acid: l‐glutamine and choline chloride: lactic acid were inefficient to recover the enzyme activity. In contrast, choline chloride: ethylene glycol: imidazole, choline chloride: glycerol: imidazole, imidazole: betaine: ethylene glycol were found to be less effective mixtures in preventing enzyme aggregation. Totally, it was demonstrated that the protective effects of the mixtures were improved as their concentrations increased. The improvement was more remarkable for imidazolium salt: β‐cyclodextrin and choline chloride: urea, where the denatured lysozyme was reactivated and recovered up to 85% of its initial activity by enhancing their concentrations from 1 to 5% (V/V). It is suggested that such solution additives may be further employed as artificial chaperones to assist protein folding and stability. [ABSTRACT FROM AUTHOR]

Background: Autolysate products from yeast origin are very interesting for food, feed, cosmetic, pharmaceutical, and fermentation industries. The lysis process greatly influences the quality and efficiency of the final autolysates., Objectives: Here, we have compared four lysis methods based on autolysis, plasmolysis (with ethanol 1.5% (v/v) and coconut fatty acids 1% (w/w)) and hydrolysis (with alkaline protease 0.4 % (v/w)) on degrading the baker's yeast Saccharomyces cerevisiae ., Materials and Methods: The efficiency of processes was evaluated according to the recovered solid and protein contents, release of intracellular materials, cell viability, microscopy imaging, degree of hydrolysis and electrophoresis studies., Results: Results showed that the increased recovered solids and proteins, as well as a higher degree of hydrolysis (DH) were obtained for the enzymatic hydrolyzed cells using alkaline protease. SDS-PAGE analysis also confirmed the results. Further, functionality of the final products by agglutination test showed that the hydrolyzed cells could effectively bind pathogenic bacteria compared to the other cell lysates., Conclusions: In conclusion, this work provides adequate evidence for efficiency of alkaline protease for producing the nutritional cell lysates from baker's S. cerevisiae used in food, feed, cosmetic, and pharmaceutical applications. Moreover, this was the first report on using coconut fatty acids and alkaline protease in lysis of baker's yeast., (Copyright: © 2021 The Author(s); Published by Iranian Journal of Biotechnology.)