An integrated omic approach towards the metabolic engineering of myrcene pathway of pseudomonas sp. M1

Best Poster AwardPseudomonas sp. M1 is able to utilize a large variety of toxic and/or recalcitrant compounds as sole carbon and energy sources, including phenols, benzene and monoterpenes like myrcene [1-3]. Therefore, M1 strain holds great potential as a source of novel biomolecules and cell factories for various biotechnological applications namely in biocatalysis, biosensors, bioremediation and biomedicine. However, the full exploitation of its enzymatic repertoire requires detailed and integrated information about the biomolecular catalog of M1 strain, including genes, proteins and metabolites. In this context, the genome of Pseudomonas sp. M1 was sequenced by NGS technologies, using Illumina GA IIx and Roche 454 FLX. The resulting raw data was assembled and annotated using different pipelines. The current genome draft of Pseudomonas sp. M1 has an estimated GC content of 67%, a size of about 7.1 Mbps and includes 6276 CDS. Importantly, in silico genome analysis predicted a number of metabolic pathways involved in utilization/biotransformation of several unusual carbons sources (e.g. biphenyls, halophenols and different monoterpenes). Proteomic and transcriptomic approaches have been setup envisaging the elucidation of the myrcene stimulon. In 2009, a set of myrcene-dependent proteins has been described using subproteome analysis of the cytoplasmic fraction [3]. In this work, a RNA-seq transcriptome analysis led to the identification of a 28kb genomic island of key importance in the catabolism of myrcene. This island includes genes involved in: i) myrcene oxidation and bioconversion of myrcene derivatives via a beta-oxidation like pathway; ii) regulation of myrcene pathway; iii) myrcene sensing. In addition several other gene clusters spread in the genome of Pseudomonas sp. M1 have been found to be myrcene-dependently expressed and are under investigation. Integration of genomic, transcriptomic, proteomic and metabolic data will deliver a very solid and detailed description of the myrcene catabolism (and other monoterpenes), and on the associated molecular mechanisms of adaptation, providing the adequate support for the application of M1 as a biocatalyst in whole-cell biotransformations of plantderived volatiles.Fundação para a Ciência e a Tecnologia (FCT

Towards the metabolic engineering of myrcene pathway of pseudomonas sp. M1 using an integrated omic approach

Pseudomonas sp. M1, isolated from the Rhine River, is able to utilize a large variety of toxic and/or recalcitrant compounds as sole carbon and energy sources, including phenols, benzene and monoterpenes like myrcene [1-3]. Therefore, M1 strain holds great potential as a source of novel biomolecules and cell factories for various biotechnological applications namely in biocatalysis, biosensors, bioremediation and biomedicine. However, the full exploitation of its enzymatic repertoire requires detailed and integrated information about the biomolecular catalog of M1 strain, including genes, proteins and metabolites. In this context, the genome of Pseudomonas sp. M1 was sequenced by NGS technologies, using Illumina Genome Analyser IIx and Roche 454 FLX. The resulting raw data was assembled into 41 contigs and annotated using different pipelines. The current genome draft of Pseudomonas sp. M1 has an estimated GC content of 67%, a size of about 6.9 Mbps and includes 6214 CDS. Importantly, in silico genome analysis predicted a number of metabolic pathways involved in utilization/biotransformation of several unusual carbons sources (e.g. biphenyls, halophenols and different monoterpenes). Proteomic and transcriptomic approaches have been setup envisaging the elucidation of the myrcene stimulon. In 2009, a set of myrcene-dependent proteins has been described using subproteome analysis of the cytoplasmic fraction [3]. More recently, a RNA-seq transcriptome analysis led to the identification of a 28kb genomic island of key importance in the catabolism of myrcene. This island includes genes involved in: i) myrcene oxidation and bioconversion of myrcene derivatives via a beta-oxidation like pathway; ii) regulation of myrcene pathway; iii) myrcene sensing. In addition several other gene clusters spread in the genome of Pseudomonas sp. M1 have been found to be myrcene-dependently expressed and are currently being characterized. Integration of genomic, transcriptomic, proteomic and metabolic data (which is currently being setup) will deliver a very solid and detailed description of the myrcene catabolism (and other monoterpenes), and on the associated molecular mechanisms of adaptation, providing the adequate support for the application of M1 as a biocatalyst in whole-cell biotransformations of plant-derived volatiles.Fundação para a Ciência e a Tecnologia (FCT

Current advances in the bacterial toolbox for the biotechnological production of monoterpene-based aroma compounds

Monoterpenes are plant secondary metabolites, widely used in industrial processes as precursors of important aroma compounds, such as vanillin and (−)-menthol. However, the physicochemical properties of monoterpenes make difficult their conventional conversion into value-added aromas. Biocatalysis, either by using whole cells or enzymes, may overcome such drawbacks in terms of purity of the final product, ecological and economic constraints of the current catalysis processes or extraction from plant material. In particular, the ability of oxidative enzymes (e.g., oxygenases) to modify the monoterpene backbone, with high regio- and stereo-selectivity, is attractive for the production of “natural” aromas for the flavor and fragrances industries. We review the research efforts carried out in the molecular analysis of bacterial monoterpene catabolic pathways and biochemical characterization of the respective key oxidative enzymes, with particular focus on the most relevant precursors, β-pinene, limonene and β-myrcene. The presented overview of the current state of art demonstrates that the specialized enzymatic repertoires of monoterpene-catabolizing bacteria are expanding the toolbox towards the tailored and sustainable biotechnological production of values-added aroma compounds (e.g., isonovalal, α-terpineol, and carvone isomers) whose implementation must be supported by the current advances in systems biology and metabolic engineering approaches.This work was supported by the project VALEU (PTDC/EAM-AMB/30488/2017); by the strategic program UID/BIA/04050/2019 through the Fundação para a Ciência e a Tecnologia (FCT) I.P.; and by the European Regional Development Fund (ERDF) through the COMPETE2020-Programa Operacional Competitividade e Internacionalização (POCI). The work was also supported by a Ph.D grant (grant number PD/BD/146184/2019) to F.S

Radon concentration assessment in water sources of public drinking of Covilhã's county, Portugal

Radon, the heaviest of the noble gases on the periodic table of elements, is a natural radioactive element that can be found on water, soils and rocks. The main goal of this work is to present an evaluation of radon concentration on samples of water, used for human consumption, collected on uranium-rich granitic rock areas. Once the geological features of the sampling region evidence the presence of this natural radionuclides, their slow dissolution steadily increases concentration in ground water. Although, the most important contribution of natural radiation, for most populations, is from inhaled radon (generic term used commonly to refer to the isotope 222Rn), in some circumstances, exposure to natural radionuclides, through drinking water, could exceed acceptable levels, and also present a hazard. Despite the fact that radon can be reduced if the water is boiled, this gas, dissolved in ground water, can be released into the air during household activities such as showering, dishwashing and laundry. So, the short lived radon decay products will contribute to increase the number of those which are present in particles suspended in the indoor air and can be accumulated up to dangerous concentrations. Once the radon progeny emits highly ionizing alpha-radiation, they may cause substantial health damage after long-term exposure. Radon concentration measurements were performed on thirty three samples collected from water wells at different depths and types of aquifers, at Covilhã's County, Portugal with the radon gas analyser DURRIDGE RAD7. Twenty three, of the total of water samples collected, gave, values over 100 Bq/L, being that 1690 Bq/L was the highest measured value.info:eu-repo/semantics/publishedVersio

Bioprospection of the bacterial β-myrcene-biotransforming trait in the rhizosphere

The biocatalysis of β-myrcene into value-added compounds, with enhanced organoleptic/therapeutic properties, may be performed by resorting to specialized enzymatic machinery of β-myrcene-biotransforming bacteria. Few β-myrcene-biotransforming bacteria have been studied, limiting the diversity of genetic modules/catabolic pathways available for biotechnological research. In our model Pseudomonas sp. strain M1, the β-myrcene catabolic core-code was identified in a 28-kb genomic island (GI). The lack of close homologs of this β-myrcene-associated genetic code prompted a bioprospection of cork oak and eucalyptus rhizospheres, from 4 geographic locations in Portugal, to evaluate the environmental diversity and dissemination of the β-myrcene-biotransforming genetic trait (Myr+). Soil microbiomes were enriched in β-myrcene-supplemented cultures, from which β-myrcene-biotransforming bacteria were isolated, belonging to Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, and Sphingobacteriia classes. From a panel of representative Myr+ isolates that included 7 bacterial genera, the production of β-myrcene derivatives previously reported in strain M1 was detected in Pseudomonas spp., Cupriavidus sp., Sphingobacterium sp., and Variovorax sp. A comparative genomics analysis against the genome of strain M1 found the M1-GI code in 11 new Pseudomonas genomes. Full nucleotide conservation of the β-myrcene core-code was observed throughout a 76-kb locus in strain M1 and all 11 Pseudomonas spp., resembling the structure of an integrative and conjugative element (ICE), despite being isolated from different niches. Furthermore, the characterization of isolates not harboring the Myr+-related 76-kb locus suggested that they may biotransform β-myrcene via alternative catabolic loci, being thereby a novel source of enzymes and biomolecule catalogue for biotechnological exploitation. KEY POINTS: • The isolation of 150 Myr+ bacteria hints the ubiquity of such trait in the rhizosphere. • The Myr+ trait is spread across different bacterial taxonomic classes. • The core-code for the Myr+ trait was detected in a novel ICE, only found in Pseudomonas spp.Open access funding provided by FCT|FCCN (b-on). This work was supported by the project VALEU (PTDC/EAM-AMB/30488/2017), by the strategic program UID/BIA/04050/2013 (POCI-01–0145-FEDER-007569), by the GenomePT project (POCI-01–0145-FEDER-022184), supported by COMPETE 2020—Opera tional Programme for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Programme (Lisboa2020), Algarve Portugal Regional Operational Programme (CRESC Algarve2020), under the Portugal 2020 Partnership Agreement, funded by national funds through the Fundação para a Ciência e a Tecnologia (FCT) I.P. and the European Regional Development Fund (ERDF), through the COMPETE2020-Programa Operacional Competitividade e Internacionalização (POCI). The work was supported as well through the Ph.D. grants to P.S-C (grant number SFRH/BD/76894/2011) and to F.S. (grant number PD/BD/146184/2019)

Synchronism in electoral cycles : how united are the United States?

The role of national, sectional, state, and local forces in driving electoral outcomes in the United States has remained a matter of considerable indeterminacy in the American politics literature. In what concerns House elections, different approaches and methods have yielded widely divergent results. In what concerns presidential elections, considerable doubts remain about the timing and the plausible causes of a long-term trend towards homogeneity. In this paper, we take a new look at the nationalization of politics in the United States. We are particularly interested in the dynamic nationalization in presidential elections, i.e., the extent to which swings and shifts from one election to the next have been similar across states and whether or not that similarity has increased through time. We treat this problem as one of similarity or dissimilarity — and convergence or divergence of — electoral cycles, and use wavelets analysis in order to ascertain the degree to which the national and state election cycles have been synchronized and the degree to which that synchronization has increased or decreased. We determine, first, the states where electoral change has been more in sync with the national cycle and clusters of states defined in terms of the mutual synchronization of their own electoral cycles. Second, we analyze how the degree of synchronization of electoral cycles in the states has changed through time, answering questions as to when, to what extent, and where has the tendency towards a “universality of political trends” in presidential elections been more strongly felt. We present evidence strongly in favor of an increase in the dynamic nationalization of presidential elections taking place in the 1950s, showing that alternative interpretations concerning the historical turning point in this respect are not supported by empirical evidence.Fundação para a Ciência e a Tecnologia (FCT

ANN based day-ahead spinning reserve forecast for electricity market simulation

Electricity market players operating in a liberalized environment require adequate decision support tools, allowing them to consider all the business opportunities and take strategic decisions. Ancillary services represent a good negotiation opportunity that must be considered by market players. This paper deals with short-term predication of day-ahead spinning reserve (SR) requirement that helps the ISO to make effective and timely decisions. Based on these forecasted information, market participants can use strategic bidding for day-ahead SR market. The proposed concepts and methodologies are implemented in MASCEM, a multi-agent based electricity market simulator. A case study based on California ISO (CAISO) data is included; the forecasted results are presented and compared with CAISO published forecast

Body composition comparison between gender in institutionalized elderly

Este estudo teve como objetivo averiguar sobre a existência de diferenças entre sexos, da composição corporal de idosos institucionalizados.The purpose of this Study was to compare the body composition differences between gender in institutionalized elderly.info:eu-repo/semantics/publishedVersio

Do the strength levels predict the motor coordination in young basketball players?

O objetivo deste estudo foi averiguar sobre a existência de relação entre os níveis de força e a coordenação motora em basquetebolistas pertencentes ao escalão de sub-18.The aim of this study was to analyse the relation between the strength levels and motor coordination in young basketball players.info:eu-repo/semantics/publishedVersio

Do the strength levels predict the motor coordination in young basketball players?

The aim of this study was to analyse the relation between the strength levels and motor coordination in young basketball players. The sample of this study was composed by 12 basketball players with 15.83 (± 0.55) years old from the U-18 category. The players participate in the inter-regional, national championship round and national cup in 2015/2016 season. The upper limbs strength was evaluated with the push-up exercise, counting the number of repetitions in 30 seconds. The lower limbs strength was evaluated in centimetres with the horizontal jump without preparatory sprint. The motor coordination was evaluated with 6 cones separated by 1,5m in the diagonal. The athletes with ball, skirted the cones and ended with layup finalization, this exercise was measured in seconds. Spearman correlation test was accessed with a significance level of 5%. No significant correlation was observed between upper limbs strength and motor coordination (F =-0,259; p =0.208). However, a negative significant correlation was observed between lower limbs strength and coordination was observed (F =-0,539; p =0,035). Thus, basketball coaches should perform specific strength training seasons intending to improve the motor coordination. Improving strength levels may contribute for motor coordination improvement.info:eu-repo/semantics/publishedVersio