Phosphoryl group wires stabilize pathological tau fibrils as revealed by multiple quantum spin counting NMR.

Hyperphosphorylation of the protein tau is one of the biomarkers of neurodegenerative diseases in the category of tauopathies. However, the molecular level, mechanistic, role of this common post-translational modification (PTM) in enhancing or reducing the aggregation propensity of tau is unclear, especially considering that combinatorial phosphorylation of multiple sites can have complex, non-additive, effects on tau protein aggregation. Since tau proteins stack in register and parallel to elongate into pathological fibrils, phosphoryl groups from adjacent tau strands with 4.8 Å separation must find an energetically favorable spatial arrangement. At first glance, this appears to be an unfavorable configuration due to the proximity of negative charges between phosphate groups from adjacent neighboring tau fibrils. However, this study tests a counterhypothesis that phosphoryl groups within the fibril core-forming segments favorably assemble into highly ordered, hydrogen-bonded, one-dimensionally extended wires under biologically relevant conditions. We selected two phosphorylation sites associated with neurodegeneration, serine 305 (S305p) and tyrosine 310 (Y310p), on a model tau peptide jR2R3-P301L (tau295-313) spanning the R2/R3 splice junction of tau, that readily aggregate into a fibril with characteristics of a seed-competent mini prion. Using multiple quantum spin counting (MQ-SC) by 31P solid-state NMR of phosphorylated jR2R3-P301L tau peptide fibrils, enhanced by dynamic nuclear polarization, we find that at least six phosphorous spins must neatly arrange in 1D within fibrils or in 2D within a protofibril to yield the experimentally observed MQ-coherence orders of four. We found that S305p stabilizes the tau fibrils and leads to more seeding-competent fibrils compared to jR2R3 P301L or Y310p. This study introduces a new concept that phosphorylation of residues within a core forming tau segment can mechanically facilitate fibril registry and stability due a hitherto unrecognized role of phosphoryl groups to form highly ordered, extended, 1D wires that stabilize pathological tau fibrils.

A comparative study to investigate the individual contribution of metabolic and physical interaction on volatiles formation in the mixed fermentation of Torulaspora delbrueckii and Saccharomyces cerevisiae.

It is well-known that the co-inoculation of Saccharomyces cerevisiae and non-Saccharomyces strains can modulate and improve the aromatic quality of wine through their multi-level interactions. However, the individual contribution of metabolic interaction (MI) and physical interaction (PI) on wine volatiles remains poorly understood. In this work, we utilized a double-compartment bioreactor to examine the aromatic effect of MI and PI by comparing the volatiles production in Torulaspora delbrueckii and Saccharomyces cerevisiae single fermentations to their mixed fermentations with or without physical separation. Results showed that the PI between T. delbrueckii and S. cerevisiae increased the production of most aroma compounds, especially for acetate esters and volatile fatty acids. In comparison, the MI only promoted a few volatile compounds, including ethyl decanoate, isoamyl acetate, and isobutanol. Noticeably, the MI significantly decreased the levels of ethyl dodecanoate, 2-phenylethyl alcohol, and decanoic acid, which exhibited opposite profiles in PI. Our results indicated that the PI was mainly responsible for the improved volatiles in T. delbrueckii/S. cerevisiae mixed fermentation, while the MI can be targeted to modulate the specific aroma compounds. A thorough understanding of the PI and MI aromatic effect will empower winemakers to accurately and directionally control the volatile profile of the wine, promoting the application of multi-starters to produce diverse styles of wines.

Real-space density kernel method for Kohn-Sham density functional theory calculations at high temperature.

Kohn-Sham density functional theory calculations using conventional diagonalization based methods become increasingly expensive as temperature increases due to the need to compute increasing numbers of partially occupied states. We present a density matrix based method for Kohn-Sham calculations at high temperatures that eliminates the need for diagonalization entirely, thus reducing the cost of such calculations significantly. Specifically, we develop real-space expressions for the electron density, electronic free energy, Hellmann-Feynman forces, and Hellmann-Feynman stress tensor in terms of an orthonormal auxiliary orbital basis and its density kernel transform, the density kernel being the matrix representation of the density operator in the auxiliary basis. Using Chebyshev filtering to generate the auxiliary basis, we next develop an approach akin to Clenshaw-Curtis spectral quadrature to calculate the individual columns of the density kernel based on the Fermi operator expansion in Chebyshev polynomials and employ a similar approach to evaluate band structure and entropic energy components. We implement the proposed formulation in the SPARC electronic structure code, using which we show systematic convergence of the aforementioned quantities to exact diagonalization results, and obtain significant speedups relative to conventional diagonalization based methods. Finally, we employ the new method to compute the self-diffusion coefficient and viscosity of aluminum at 116 045 K from Kohn-Sham quantum molecular dynamics, where we find agreement with previous more approximate orbital-free density functional methods.

Effects of three indigenous non-Saccharomyces yeasts and their pairwise combinations in co-fermentation with Saccharomyces cerevisiae on volatile compounds of Petit Manseng wines.

The combined use of selected Saccharomyces cerevisiae and non-Saccharomyces strains is becoming an effective way to achieve wine products with distinctive aromas. The purpose of this study was to further improve the wine aroma complexity through optimizing inoculation protocols of multi-starters. The three indigenous non-Saccharomyces strains (Torulaspora delbrueckii, Hanseniaspora vineae, and Lachancea thermotolerans) and their pairwise combinations (co-inoculation) were sequentially inoculated with S. cerevisiae in Petit Manseng grape must, respectively. Results evidenced a higher divergence in aroma compounds produced by two different non-Saccharomyces species compared to single species. Especially for the combination of T. delbrueckii and L. thermotolerans, the concentrations of most ethyl esters were further increased, contributing to a higher score of 'pineapple' note in agreement with sensory analysis. Our results highlighted that the inoculation of more than one non-Saccharomyces species is a potential strategy to improve the aroma diversity and quality of industrial wines.

Use of Indigenous Hanseniaspora vineae and Metschnikowia pulcherrima Co-fermentation With Saccharomyces cerevisiae to Improve the Aroma Diversity of Vidal Blanc Icewine.

Using novel non-Saccharomyces strains is regarded as an effective way to improve the aroma diversity of wines to meet the expectations of consumers. The non-Saccharomyces Hanseniaspora vineae and Metschnikowia pulcherrima have good aromatic properties useful for the production of table wine. However, no detailed information is available on their performances in icewine fermentation. In this study, simultaneous and sequential fermentation trials of indigenous M. pulcherrima CVE-MP20 or H. vineae CVE-HV11 with S. cerevisiae (SC45) were performed in 50-L fermenters of Vidal icewine, respectively. The results showed that SC45 cofermented with different non-Saccharomyces strains could generate a distinct aroma quality of icewine compared with four S. cerevisiae strain monocultures as evidenced by principal component analysis (PCA). Mixed fermentation of MP20/SC45 produced higher contents of acetate esters and ß-damascenone with lower C6 alcohols relative to SC45 monoculture. Interestingly, HV11/SC45 generated the highest amounts of C6 alcohols [(Z)-3-hexen-1-ol and (E)-3-hexen-1-ol], higher alcohols (isobutanol, isopentanol, and 2-phenylethanol), acetate esters (2-phenethyl acetate and isoamyl acetate), cis-rose oxide, ß-damascenone, and phenylacetaldehyde. Compared with simultaneous inoculation, sequential inoculation could achieve higher aroma diversity and produce higher intensity of fruity, flowery, and sweet attributes of icewine as assessed by calculating the odor activity values (OAVs). Our results verified the desired enological characteristics of H. vineae strain in icewine fermentation and also demonstrated that using indigenous non-Saccharomyces and Saccharomyces strains is a feasible way to improve aroma diversity of icewine products, which could provide an alternative way to meet the requirement of wine consumers for diversified aromatic quality.

Use of Torulaspora delbrueckii Co-fermentation With Two Saccharomyces cerevisiae Strains With Different Aromatic Characteristic to Improve the Diversity of Red Wine Aroma Profile.

The use of selected Saccharomyces and non-Saccharomyces strains as mixed starters has advantages over pure fermentation due to achieving wine products with distinctive and diversified aroma expected by consumers. To obtain a way to improve the aroma diversity and increase the differentiation of wine product, in this study, the aromatic effect of multi-culture of indigenous Torulaspora delbrueckii (TD12), simultaneous and sequential inoculation with two Saccharomyces strains (indigenous icewine yeast SC45 and commercial yeast BDX) with different enological characteristics were investigated in laboratory-scale 20 L fermenter, respectively. The results showed that T. delbrueckii co-fermented with different S. cerevisiae strain could generate diversified physicochemical and aromatic quality of wine as evidenced by PCA. Mixed fermentation of SC45/TD12 produced higher contents of higher alcohol (3-methyl-1-pentanol and phenylethyl alcohol), ethyl esters (ethyl decanoate and ethyl butanoate), terpenes and phenylacetaldehyde with less fatty acids (hexanoic acid, octanoic acid) and acetic acid, while BDX/TD12 generated more C6 alcohol (1-hexanol) and acetate esters (ethyl acetate and isoamyl acetate). Compared to simultaneous inoculation, sequential inoculation could achieve higher aroma diversity, and generate higher intensity of fruity, flowery and sweet attributes of wine as assessed by calculating the odor activity values. The different S. cerevisiae strain and inoculation method in alcoholic fermentation could further influence the formations of aromatic compounds in malolactic fermentation. Our results highlighted the importance of S. cerevisiae strain in shaping the aromatic quality of wine in mixed fermentation, and also suggested that using different S. cerevisiae strains with distinct aromatic characteristics co-fermentation with specific non-Saccharomyces strain is a potential way to increase the aromatic diversity and quality of wine product, which could provide an alternative way to meet the requirement of wine consumers for diversified aromatic quality.