Endo-1,5-α-L-arabinanases are glycosyl hydrolases that are able to cleave the glycosidic bonds of α-1,5-L-arabinan, releasing arabino-oligosaccharides and L-arabinose. Two extracellular endo-1,5-α-L-arabinanases have been isolated from Bacillus subtilis, BsArb43A and BsArb43B (formally named AbnA and Abn2, respectively). BsArb43B shows low sequence identity with previously characterized 1,5-α-L-arabinanases and is a much larger enzyme. Here we describe the 3D structure of native BsArb43B, biochemical and structure characterization of two BsArb43B mutant proteins (H318A and D171A), and the 3D structure of the BsArb43B D171A mutant enzyme in complex with arabinohexose. The 3D structure of BsArb43B is different from that of other structurally characterized endo-1,5-α-L-arabinanases, as it comprises two domains, an N-terminal catalytic domain, with a 3D fold similar to that observed for other endo-1,5-α-L-arabinanases, and an additional C-terminal domain. Moreover, this work also provides experimental evidence for the presence of a cluster containing a calcium ion in the catalytic domain, and the importance of this calcium ion in the enzymatic mechanism of BsArb43B.

Bacillus subtilis is able to utilize arabinopolysaccharides derived from plant biomass. Here, by combining genetic and physiological analyses we characterize the AraNPQ importer and identify primary and secondary transporters of B. subtilis involved in the uptake of arabinosaccharides. We show that the ABC-type importer AraNPQ is involved in the uptake of α-1,5-arabinooligosaccharides, at least up to four L-arabinosyl units. Although this system is the key transporter for α-1,5-arabinotriose and α-1,5-arabinotetraose, the results indicate that α-1,5-arabinobiose also is translocated by the secondary transporter AraE. This broad-specificity proton symporter is the major transporter for arabinose and also is accountable for the uptake of xylose and galactose. In addition, MsmX is shown to be the ATPase that energizes the incomplete AraNPQ importer. Furthermore, the results suggest the existence of at least one more unidentified MsmX-dependent ABC importer responsible for the uptake of nonlinear α-1,2- and α-1,3-arabinooligosaccharides. This study assigns MsmX as a multipurpose B. subtilis ATPase required to energize different saccharide transporters, the arabinooligosaccharide-specific AraNPQ-MsmX system, a putative MsmX-dependent ABC transporter specific for nonlinear arabinooligosaccharides, and the previously characterized maltodextrin-specific MdxEFG-MsmX system.

The solid-phase synthesis (SPS) of a structurally complex glycopeptide, using Sieber amide resin, was monitored by high resolution magic angle spinning NMR, demonstrating the further application of this technique. A synthetic peptidoglycan derivative, a precursor of a biologically active PGN, known to be involved in the cellular recognition, was prepared by SPS. The synthesis involved the preparation of an N-alloc glucosamine moiety and the synthesis of a simple amino acid sequence L-Ala-D-Glu-L-Lys-D-Ala-D-Ala. Last step consisted the coupling, on solid-phase, of the protected muramyl unit to the peptide chain. Proton spectra with good suppression of the polystyrene signals in swollen resin samples were obtained in DMF-d(7) as a solvent and by using a nonselective 1D TOCSY/DIPSI-2 scheme, thus allowing to follow the SPS without losses of compound and cleavage from the resin. The assignment of the proton spectra of the resin-bound amino acid sequence and of the bound glycopeptide was achieved through the combination of MAS COSY, TOCSY and NOESY.

Molecularly imprinted polymers (MIPs) of poly(ethylene glycol dimethacrylate) and poly(N-isopropylacrylamide-co-ethylene glycol dimethacrylate) were synthesized for the first time in supercritical carbon dioxide (scCO2), using Boc-l-tryptophan as template. Supercritical fluid technology provides a clean and one-step synthetic route for the preparation of affinity polymeric materials with sensing capability for specific molecules. The polymeric materials were tested as stationary \{HPLC\} phases for the enantiomeric separation of l- and d-tryptophan. \{HPLC\} results prove that the synthesized \{MIPs\} are able to recognize the template molecule towards its enantiomer which opens up potential applications in chromatographic chiral separation.

Sleep staging is a crucial step before the scoring the sleep apnoea, in subjects that are tested for this condition. These patients undergo a whole night polysomnography recording that includes EEG, EOG, ECG, EMG and respiratory signals. Sleep staging refers to the quantification of its depth. Despite the commercial sleep software being able to stage the sleep, there is a general lack of confidence amongst health practitioners of these machine results. Generally the sleep scoring is done over the visual inspection of the overnight patient EEG recording, which takes the attention of an expert medical practitioner over a couple of hours. This contributes to a waiting list of two years for patients of the Portuguese Health Service. In this work we have used a spectral comparison method called Itakura distance to be able to make a distinction between sleepy and awake epochs in a night EEG recording, therefore automatically doing the staging. We have used the data from 20 patients of Hospital Pulido Valente, which had been previously visually expert scored. Our technique results were promising, in a way that Itakura distance can, by itself, distinguish with a good degree of certainty the N2, N3 and awake states. Pre-processing stages for artefact reduction and baseline removal using Wavelets were applied.

Deuterium NMR was used to investigate the orientational order in a composite cellulosic formed by liquid crystalline acetoxypropylcellulose (A PC) and demented nematic 4'-penty1-4-cyanobiphenyl (5CB-4 alpha d(2)) with the per centage of 85% A PC by weight Three forms of the composite including electro spun microfibers, thin film and bulk samples were analyzed The NMR results initially suggest two distinct scenarios, one whet e the 503-alpha d(2), is confined to small droplets with dimensions smaller than the magnetic coherence length and the other where the 503-alpha d(2) molecules arc aligned with the A PC network chains Polarized optical microscopy (POW from thin film samples along with all the NMR results show the presence of 5CB-alpha d(2) droplets in the composite systems with a nematic wetting layer at the APC-5CB-alpha d(2) interface that experiences and order disorder transition driven by the polymer network N-I transition The characterization of the APC network I-N transition shows a pronounced subcritical behavior within a heterogeneity scenario

The design and optimization of fractional filters is considered in this paper. Some of the classic filter architectures are presented and their performances relatively to an ideal amplitude spectrum evaluated. The fractional filters are designed using the differential evolution optimization algorithm for computing their parameters. To evaluate the performances of all the filters the quadratic error between the computed amplitude is calculated against an ideal (goal) response. The fractional filters have a better behavior, both in the pass and reject-band.

The quantum fractional derivative is defined using formulations analogue to the common Grünwald?Letnikov derivatives. While these use a linear variable scale, the quantum derivative uses an exponential scale and is defined in R? or R?. Two integral formulations similar to the usual Liouville derivatives are deduced with the help of the Mellin transform.

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