Multivalent enzymes that enable the use of hydrogen peroxide for microaerobic and anaerobic proliferation

This work was financed by national funds from FCT - Fundação para a Ciência e a Tecnologia, I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences - UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy - i4HB. FCT supported SRP through the projects PTDC/BIA-PRO/109796/2009 and PTDC/BIA-BQM/29442/2017, DSB and RNSO through the scholarships UI/BD/151168/2021, and “Verão com Ciência2020”, respectively.Bacterial peroxidases are responsible for the reduction of hydrogen peroxide to water. Found in the periplasm of gram-negative bacteria, they are one of the defense mechanisms against endogenous and exogenous peroxide stress under low oxygen tensions. Besides being involved in peroxide detoxification, bacterial peroxidases have been proposed to constitute an alternative pathway to the respiratory chain under anoxic conditions, as demonstrated in E. coli that can use hydrogen peroxide as an electron acceptor in the absence of oxygen. Bacterial peroxidases are c-type cytochromes with either two or three c-type hemes bound to the polypeptide chain, being divided into classical or non-classical, respectively. Orthologous to the classical bacterial peroxidases are the MauG enzymes that share some structural, spectroscopic and sequence similarities but have distinct physiological roles (though for most their function remains unknown). The spectroscopic and kinetic data on bacterial peroxidases are reviewed for both classes. Most classical bacterial peroxidases require reductive activation that consists in structural changes so that the catalytic heme becomes accessible to the substrate. However, non-classical enzymes are ready to bind the hydrogen peroxide as their catalytic center is penta-coordinated, which is also observed in their structural model. The few studies that report the involvement of bacterial peroxidases from pathogenic bacteria in biofilms, is an indication that these enzymes might contribute to their infection mechanism and thus can constitute alternative drug targetspublishersversionpublishe

Bacteriocin production by Lactobacillus plantarum ST16Pa in supplemented whey powder formulations

Whey, the main by-product of the dairy industry, is frequently disposed of in the environment without any treatment due to the high cost of this process. Alternatively, whey can be used as a medium to culture lactic acid bacteria and produce value-added products such as bacteriocins. In this work, we attempted to improve bacteriocin production by Lactobacillus plantarum ST16Pa in a whey powder formulation supplemented with additional sources of carbon, nitrogen, and vitamin B12 at different levels and varying the agitation intensity according to a Plackett-Burman experimental design. Only the addition of tryptone positively influenced the production of this bacteriocin. The results allowed us to identify a supplemented whey formulation, comprising 150 g/L of whey total solids plus 10 g/L of tryptone and soybean extract, whose fermentation by Lb. plantarum ST16Pa in shake flasks under agitation at 150 rpm led to a cell-free supernatant with an antimicrobial activity against Listeria innocua 6a CLIST 2865 (inhibition zone of 13.23 mm) close to that previously obtained in de Man, Rogosa and Sharpe medium by other authors. These results are significant considering that the same strain cultured in cheese whey did not previously display any antimicrobial activity

Mechanical Evaluation of Polymer Composite Hip Protectors

Hip fractures often result in serious health implications, particularly in the geriatric population, and have been related to long-term morbidity and death. In most cases, these fractures are caused by impact loads in the area of the greater trochanter, which are produced in a fall. This work is aimed at developing hip protectors using composite materials and evaluating their effectiveness in preventing hip fractures under high impact energy (120 J). The hip protectors were developed with an inner layer of energy absorbing soft material and an outer rigid shell of fiberglass-reinforced polymer composite. According to the experimental results, all tested configurations proved to be effective at reducing the impact load to below the average fracture threshold of proximal femur. Furthermore, an addition of Ethylene Vinyl Acetate (EVA) to the impacted area of the composite shell proved to be beneficial to increase impact strength of the hip protectors. Thus, composite hip protectors proved to be a viable alternative for a mechanically efficient and cost-effective solution to prevent hip fractures

Application of chemometric tools for the comparison of volatile profile from raw and roasted regional and foreign almond cultivars (Prunus dulcis)

In almonds, volatile compounds are major contributors to flavour, being scarce the current knowledge about their volatile profile. Hence, this work intended to characterize the volatile profile, using headspace solidphase microextraction and gas chromatography–mass spectrometry, in raw and roasted almond cultivars (regional cvs. Amendoa˜o, Bonita, Casanova, Molar and Pegarinhos and foreign cvs. Ferragne`s and Glorieta). Overall, 35 compounds were identified, with major chemical classes being alcohols and aldehydes. In raw fruits, benzaldehyde and 3-methyl-1-butanol were key compounds, with roasting changing volatile profiles, increasing release of compounds, with predominance of hexanal and benzaldehyde. Cultivars Glorieta and Molar didn’t show significant increase in aldehyde content after roasting, which may indicate higher resistance to heat-caused oxidation. The use of linear discriminant analysis and principal components analysis permitted the recognition of patterns in the volatile profiles, that can be useful for cultivars identification. This work allowed the characterization and monitoring changes caused by roasting of volatile components of less studied almond cultivars, identifying some that can withstand roasting procedures with reduced formation of compounds associated with off-flavours.Ivo Oliveira is grateful to FCT, POPH-QREN and FSE for the Post-doctoral Fellowship SFRH/BPD/111005/2015. This work is supported by: European Investment Funds by FEDER/ COMPETE/POCI—Operational Competitiveness and Internacionalization Programme, under Project POCI-01-0145-FEDER-006958 and National Funds by FCT—Portuguese Foundation for Science and Technology, under the Project UID/AGR/04033/2013.info:eu-repo/semantics/publishedVersio

Merging zones approach in a flow-based platform for the determination of the total protein content in microbiological samples

info:eu-repo/semantics/publishedVersio

Stimuli-sensitive self-assembled tubules based on lysine-derived surfactants as nanocarriers for proteins

Drug delivery vectors based on amphiphilic molecules present considerable advantages, namely versatility in physicochemical properties and sensitivity to stimuli. Amino acid-based surfactants, in particular, are rather promising amphiphiles for this purpose1 because of their enhanced biocompatibility compared to conventional surfactants. In addition to forming micelles and vesicles, they can self-organize into other complex supramolecular structures, such as fibers, twisted ribbons, helical tapes and nanotubes.2,3 Herein, we have studied a family of novel anionic double-chained lysine-based surfactants, with variable degree of chain length mismatch. Because of their peculiar structure, these compounds are able to form in water tubular structures with assorted morphologies, as evidenced by video-enhanced light microscopy (VELM), scanning electron microscopy (SEM and cryo-SEM), cryogenic transmission electron microscopy (cryo-TEM) and atomic force microscopy (AFM).3 The loading ability of the tubules towards lysozyme, under varying experimental conditions, has been investigated inter alia by differential scanning microcalorimetry, gel electrophoresis and UV/VIS spectroscopy, with the goal of assessing the efficiency of these aggregates as pH- and temperature-sensitive nanocarriers for a model biomolecule. Results on the stability of the native and loaded tubules when in contact with different fluids (serum, artificial saliva, artificial sweat, blood), and on their toxicity in human cells, are also presented and discussed.FCT is gratefully acknowledged for financial support through Ph.D. grant SFRH/BD/108629/2015. CIQUP acknowledges financial support from FEDER/COMPETE and FCT through grants UID/QUI/00081/2013, POCI-01-0145-FEDER- 006980 and NORTE-01-0145-FEDER-000028

Natural based reusable materials for microfluidic substrates: The silk road towards sustainable portable analytical systems

Portable analytical systems are versatile tools for application in areas including biomedicine, biosecurity, food safety and environmental monitoring. This work contributes to the increasing demand for low-cost, environmentally friendly substrates for portable analytical systems by using natural Bombyx mori cocoons. Further, silk fibroin is also extracted from these cocoons and electrospun into oriented and randomly oriented fiber substrates. Oxygen plasma treatment is applied to improve their hydrophilicity. Fiber morphology, mechanical properties, porosity, thermal characteristics and surface contact angle are extensively characterized and the ability of the samples for passive capillary flows demonstrated. Plasma treated pressed cocoons show superhydrophilicity, capillary flow rates of 44.8 ± 3.75 mm.min-1, and high mechanical resistance with Young's modulus values up to 592.13 ± 19.83 MPa. The developed materials are used as substrates for the colorimetric quantification of three commonly scrutinized clinical analytes. Hydrophobic barriers are first wax-printed on all samples with a proper design and albumin assays are performed on all substrates. Further assays for uric acid and glucose quantification are successfully accomplished on the pressed cocoons after a simple in between washing step, with overall high coefficient of determination, proving the suitability of the developed materials as low-cost, sustainable and reusable microfluidic substrates.This work was supported by the Portuguese Foundation for Science and Technology (FCT) under strategic funding UID/FIS/04650/2020, UIDB/04436/2020, UIDP/04436/2020 and project PTDC/EMD-EMD/28159/2017 (POCI-01–0145-FEDER-028159). The authors also thank FCT for financial support under grants SFRH/BD/140698/2018 (R.B. P.), 2020.09218.BD (A.S.M.), 2020.04163.CEECIND (C.R.) and 2020.02304.CEECIND (V.F.C.). Finally, the authors acknowledge fund ing by Spanish State Research Agency (AEI) and the European Regional Development Fund (ERFD) through the project PID2019–106099RB C43/AEI/10.13039/501100011033 and from the Basque Government Industry Departments under the ELKARTEK program. Finally, the au thors also thank Dr. J. Borges and Prof. F. Vaz for experimental support.info:eu-repo/semantics/publishedVersio

Optimal binary search trees with costs depending on the access paths

We describe algorithms for constructing optimal binary search trees, in which the access cost of a key depends on the k preceding keys which were reached in the path to it. This problem has applications to searching on secondary memory and robotics. Two kinds of optimal trees are considered, namely optimal worst case trees and weighted average case trees. The time and space complexities of both algorithms are O(nᵏ+²) and O(nᵏ+¹ ), respectively. The algorithms are based on a convenient decomposition and characterizations of sequences of keys which are paths of special kinds in binary search trees. Finally, using generating funcions, we present an exact analysis of the number of steps performed by the algorithms