Impact of transglutaminase treatment on properties and in vitro digestibility of white bean (Phaseolus vulgaris L.) flour

Common beans (Phaseolus vulgaris L.) are rich in nutrients and have significant amounts of proteins and complex carbohydrates, besides to be rich in unsaturated fatty acids and dietary fibres. Consumption of beans could be improved by processing them into flour. In this study the effect of microbial transglutaminase (TG) on the structure, physical (colour parameters, moisture, water holding capacity), thermal properties and in vitro digestion of undehulled (WB) and manually dehulled (SB) flour samples from white common beans (P. vulgaris L.) was evaluated. Flour samples were incubated in the absence and presence of TG (WB/TG and SB/TG). We observed that the enzyme is able to catalyse the formation of polymers, suggesting that the proteins occurring in the bean flour act as TG substrates. Microstructure of samples was examined by Scanning Electron Microscopy (SEM), while thermal properties were studied by Differential Scanning Calorimetry. Microstructural results showed that the TG-treated samples possess a more compact structure, made of starch granules surrounded by proteins that, presumably, contain TG-catalysed polymers. Moreover, TG treatment had a major impact on colour, water holding capacity (WHC) and thermal properties. In particular, WB and SB samples presented a darker colour than WB/TG and SB/TG samples, while the latter showed reduced WHC that was only 30% and 37% of WB and SB samples, respectively. The transition enthalpy (ΔH) in the temperature range from 57 to 70 °C (WB, WB/TG) and from 60 to 68 °C (SB, SB/TG) followed the order: WB/TG > WB and SB/TG > WB, respectively. In vitro digestion experiments indicate that the presence of isopeptide bonds decreased the digestibility of TG-treated flour samples

Effect of the refreshment on the liquid sourdough preparation

The aim of this work was to investigate the effect of refreshments on the growth of endogenous microorganisms during liquid sourdough preparation by using an Italian and Mexican wheat flours and its effects on the physico-chemical properties (pH, total titratable acidity, water activity, moisture content and reducing sugars). The liquid sourdoughs were prepared (DY 200) and incubated for 6 days at 20°C. The sourdoughs were refreshed every day and compared with the not-refreshed ones. Preliminary results showed that in the early stages of the microbial growth process, their population was greater in the sourdough made from the Mexican wheat flour than that of the Italian one. However, after 6 days, the microbial population was not significantly different in refreshed or not-refreshed samples for both sourdoughs (Italian and Mexican). Similarly, physicochemical properties did not show significant differences

Search for gravitational-lensing signatures in the full third observing run of the LIGO-Virgo network

Gravitational lensing by massive objects along the line of sight to the source causes distortions of gravitational wave-signals; such distortions may reveal information about fundamental physics, cosmology and astrophysics. In this work, we have extended the search for lensing signatures to all binary black hole events from the third observing run of the LIGO--Virgo network. We search for repeated signals from strong lensing by 1) performing targeted searches for subthreshold signals, 2) calculating the degree of overlap amongst the intrinsic parameters and sky location of pairs of signals, 3) comparing the similarities of the spectrograms amongst pairs of signals, and 4) performing dual-signal Bayesian analysis that takes into account selection effects and astrophysical knowledge. We also search for distortions to the gravitational waveform caused by 1) frequency-independent phase shifts in strongly lensed images, and 2) frequency-dependent modulation of the amplitude and phase due to point masses. None of these searches yields significant evidence for lensing. Finally, we use the non-detection of gravitational-wave lensing to constrain the lensing rate based on the latest merger-rate estimates and the fraction of dark matter composed of compact objects

Search for eccentric black hole coalescences during the third observing run of LIGO and Virgo

Despite the growing number of confident binary black hole coalescences observed through gravitational waves so far, the astrophysical origin of these binaries remains uncertain. Orbital eccentricity is one of the clearest tracers of binary formation channels. Identifying binary eccentricity, however, remains challenging due to the limited availability of gravitational waveforms that include effects of eccentricity. Here, we present observational results for a waveform-independent search sensitive to eccentric black hole coalescences, covering the third observing run (O3) of the LIGO and Virgo detectors. We identified no new high-significance candidates beyond those that were already identified with searches focusing on quasi-circular binaries. We determine the sensitivity of our search to high-mass (total mass M>70 M⊙) binaries covering eccentricities up to 0.3 at 15 Hz orbital frequency, and use this to compare model predictions to search results. Assuming all detections are indeed quasi-circular, for our fiducial population model, we place an upper limit for the merger rate density of high-mass binaries with eccentricities 0<e≤0.3 at 0.33 Gpc−3 yr−1 at 90\% confidence level

Ultralight vector dark matter search using data from the KAGRA O3GK run

Among the various candidates for dark matter (DM), ultralight vector DM can be probed by laser interferometric gravitational wave detectors through the measurement of oscillating length changes in the arm cavities. In this context, KAGRA has a unique feature due to differing compositions of its mirrors, enhancing the signal of vector DM in the length change in the auxiliary channels. Here we present the result of a search for U(1)B−L gauge boson DM using the KAGRA data from auxiliary length channels during the first joint observation run together with GEO600. By applying our search pipeline, which takes into account the stochastic nature of ultralight DM, upper bounds on the coupling strength between the U(1)B−L gauge boson and ordinary matter are obtained for a range of DM masses. While our constraints are less stringent than those derived from previous experiments, this study demonstrates the applicability of our method to the lower-mass vector DM search, which is made difficult in this measurement by the short observation time compared to the auto-correlation time scale of DM

Nickel and GTP Modulate Helicobacter pylori UreG Structural Flexibility

UreG is a P-loop GTP hydrolase involved in the maturation of nickel-containing urease, an essential enzyme found in plants, fungi, bacteria, and archaea. This protein couples the hydrolysis of GTP to the delivery of Ni(II) into the active site of apo-urease, interacting with other urease chaperones in a multi-protein complex necessary for enzyme activation. Whereas the conformation of Helicobacter pylori (Hp) UreG was solved by crystallography when it is in complex with two other chaperones, in solution the protein was found in a disordered and flexible form, defining it as an intrinsically disordered enzyme and indicating that the well-folded structure found in the crystal state does not fully reflect the behavior of the protein in solution. Here, isothermal titration calorimetry and site-directed spin labeling coupled to electron paramagnetic spectroscopy were successfully combined to investigate HpUreG structural dynamics in solution and the effect of Ni(II) and GTP on protein mobility. The results demonstrate that, although the protein maintains a flexible behavior in the metal and nucleotide bound forms, concomitant addition of Ni(II) and GTP exerts a structural change through the crosstalk of different protein regions

Etude de la dynamique structurale de protéines dans des cellules bactériennes par marquage de spin et spectroscopie RPE : de l'amélioration des méthodes aux études dans les cellules

L'étude des biomolécules dans leur environnement natif, la cellule, est l'un des principaux objectifs de la biologie structurale au cours de la dernière décennie. Ainsi, nous assistons à un remarquable développement des approches dites « in-cell », comme la CryoET, FRET (Förster Resonance Energy Tranfer), RMN. Parmi elles, la technique de marquage de spin couplée à la spectroscopie de Résonance Paramagnétique Electronique (SDSL-RPE) présente des caractéristiques intéressantes et avantageuses permettant de sonder la dynamique des protéines à l'intérieur des cellules. En particulier, l’utilisation des sondes de type nitroxyde combine sensibilité élevée et absence de contraintes de taille de la biomolécule d'intérêt avec la capacité d'étudier les transitions structurales et les interactions protéines-protéines à température physiologique. Cependant, même si de nombreux efforts ont été faits pour adapter cette technique à des études structurales dans les cellules, des progrès restent à faire.Dans cette thèse, nous traitons des principales limitations de l’utilisation des nitroxydes dans un contexte cellulaire. Nous nous sommes concentrés sur la stabilité des marqueurs nitroxydes dans des milieux reducteurs et dans la cellule, sur l’incorporation des protéines marquées dans les cellules et la viabilité de ces cellules en vue de mesures par RPE. Grâce aux résultats obtenus dans cette partie méthodologique, nous avons pu étudier la dynamique structurale de deux protéines chaperons (NarJ et UreG) dans des cellules bactériennes. Ces avancées ont permis de comparer les données obtenues in-cell à celles obtenues in vitro ou dans un environnement mimant le milieu cellulaire.The study of biomolecules in their native environment has been one of the main goals of structural biology in the last decade. As a result, we are assisting to a remarkable increase of new "in-cell" approaches, like Cryo-ET, FRET and NMR. Among these approaches, Site-Directed Spin Labeling (SDSL) coupled to Electron Paramagnetic Resonance (EPR) spectroscopy shows competitive and advantageous features to capture protein dynamics inside cells. In particular, nitroxide-based SDSL-EPR combines the advantages of high sensitivity and the lack of size constraints on the biomolecule of interest with the ability to capture protein structural transitions and interactions at physiological temperature. Despite the methodological advancements of the technique that have allowed the community to obtain increasingly relevant results, progresses still need to be done.In this work, the main limitation of nitroxide-based SDSL-EPR has been addressed. In the first time, we focused on the development of delivery methods to introduce the labeled protein in bacterial cells. Next, the stability of nitroxide labels in reducing environments and in-cell has been assessed, monitoring in parallel the viability of the cells during the EPR measurements. Thanks to the results achieved in this methodological part, we were able to study the structural dynamics of two flexible chaperone proteins directly in bacterial cells: NarJ from Escherichia coli and UreG from Sporosarcina pasteurii. Finally, to go further in understanding the impact of the cellular environment on the protein dynamics, the data obtained in cellular context were compared with those obtained in vitro or in a cell-mimicking environment

Dance with spins : site-directed spin labeling coupled to electron paramagnetic resonance spectroscopy directly inside cells

Depicting how biomolecules move and interact within their physiological environment is one of the hottest topics of structural biology. This Feature Article gives an overview of the most recent advances in Site-directed Spin Labeling coupled to Electron Paramagnetic Resonance spectroscopy (SDSL–EPR) to study biomolecules in living cells. The high sensitivity, the virtual absence of background, and the versatility of spin-labeling strategies make this approach one of the most promising techniques for the study of biomolecules in physiologically relevant environments. After presenting the milestones achieved in this field, we present a summary of the future goals and ambitions of this community.publishe