Plant biomass-degrading microbial consortia:Ecology, lignocellulolytic machinery and biotechnological potential

De productie van hernieuwbare ‘bio-based’ verbindingen uit suikers verkregen uit plantaardige biomassa is de afgelopen tien jaar sterk toegenomen. Een belangrijke drempel in dergelijke processen is dikwijls de lage efficiëntie van versuikering, veroorzaakt door de recalcitrante aard van de lignocellulose matrix. Microbiële consortia zijn wel voorgesteld als aantrekkelijke alternatieven voor een efficiëntere afbraak van deze materialen. In deze studie beschrijven wij verschillende microbiële consortia die op tarwestro, maïsstro en switchgrass werden gekweekt als enige bronnen van koolstof en energie. Met name zijn metataxonomische (gestoeld op 16S rRNA gen/ ITS1 basevolgordebepalingen) en metagenomische analyses (via DNA basevolgordebepaling en screening van bibliotheken) uitgevoerd, waardoor gegevens over de gekweekte microbiële consortia werden verkregen. In deze consortia werden leden van de Enterobacteriales, Pseudomonadales, Flavobacteriales en Sphingobacteriales als dominante soorten waargenomen. Daarnaast zijn de totale uitgeschieden eiwitten onderzocht via vloeistofchromatografie-tandem massaspectrometrie, waarna de enzymatische activiteiten geëvalueerd werden met behulp van een nieuwe generatie van chromogene substraten. De ecofysiologie van de systemen kan worden gekenmerkt als verrassend complex en ingewikkeld. Bijvoorbeeld waren de consortiumsamenstellingen complex, waarbij ook is aangetoond dat de verschillende consortiumleden invloed hadden op het lignocellulose-afbraakproces. Het enzymatische complement van de consortia bleek uitgescheiden xylanasen, glucosidases, arabinofuranosidasen, galactosidasen, mannosidases en fucosidases te bevatten. Een analyse van de metagenomische bibliotheken onthulde de aanwezigheid van verschillende genen voor nieuwe hitte-alkali-tolerante enzymen. De verkregen metagenomen en metasecretomen vormen een waardevolle bron voor enzymcocktails die bruikbaar zijn in de processen voor versuikering van lignocellulose

Metasecretome analysis of a lignocellulolytic microbial consortium grown on wheat straw, xylan and xylose

Background: Synergistic action of different enzymes is required to complete the degradation of plant biomass in order to release sugars which are useful for biorefining. However, the use of single strains is often not efficient, as crucial parts of the required enzymatic machinery can be absent. The use of microbial consortia bred on plant biomass is a way to overcome this hurdle. In these, secreted proteins constitute sources of relevant enzyme cocktails. Extensive analyses of the proteins secreted by effective microbial consortia will contribute to a better understanding of the mechanism of lignocellulose degradation. Results: Here, we report an analysis of the proteins secreted by a microbial consortium (metasecretome) that was grown on either wheat straw (RWS), xylose or xylan as the carbon sources. Liquid chromatography-tandem mass spectrometry was used to analyze the proteins in the supernatants. Totals of 768 (RWS), 477 (xylose) and 103 (xylan) proteins were identified and taxonomically and functionally classified. In RWS, the proteins were mostly affiliated with Sphingobacterium-like consortium members (similar to 50 %). Specific abundant protein clusters were predicted to be involved in polysaccharide transport and/or sensing (TonB-dependent receptors). In addition, proteins predicted to degrade plant biomass, i.e. endo-1,4-beta-xylanases, alpha-l-arabinofuranosidases and alpha-l-fucosidases, were prominent. In the xylose-driven consortium, most secreted proteins were affiliated with those from Enterobacteriales (mostly Klebsiella species), whereas in the xylan-driven one, they were related to Flavobacterium-like ones. Notably, the metasecretomes of the consortia growing on xylose and xylan contained proteins involved in diverse metabolic functions (e.g. membrane proteins, isomerases, dehydrogenases and oxidoreductases). Conclusions: An analysis of the metasecretomes of microbial consortia originating from the same source consortium and subsequently bred on three different carbon sources indicated that the major active microorganisms in the three final consortia differed. Importantly, diverse glycosyl hydrolases, predicted to be involved in (hemi) cellulose degradation (e.g. of CAZy families GH3, GH10, GH43, GH51, GH67 and GH95), were identified in the RWS metasecretome. Based on these results, we catalogued the RWS consortium as a true microbial enzyme factory that constitute an excellent source for the production of an efficient enzyme cocktail for the pretreatment of plant biomass

Desarrollo de un sistema automático de colorimetría CMYK con visión artificial.

El trabajo de titulación tuvo como objetivo el desarrollo de un sistema automático de colorimetría con visión artificial en base al modelo sustractivo Cian, Magenta, Amarillo, Negro (CMYK). El sistema se implementó primeramente diseñando la estructura en un software de dibujo asistido por computadora (CAD), posteriormente se hizo la adquisición de los materiales. Se procedió al armado de la estructura, para así poder realizar la programación en un controlador lógico programable (PLC), mientras que el sistema de control se lo realizó en un interfaz hombre máquina (HMI) mediante el Software Lookout y la parte de la adquisición del color se lo efectuó en el software de programación MATLAB. Una vez realizadas las pruebas se constató que el porcentaje de error de los valores calculados por el sistema implementado respecto a los valores teóricos de los colores CMYK, son mínimos aproximadamente del 3.8%; con lo que se corrobora la eficiencia de la máquina. El sistema de colorimetría permitió asignar un valor de porcentaje para las tintas de cuatricromía y se realiza la mezcla de las mismas para obtener un color capturado por la cámara del celular. Se recomienda a los estudiantes que previo a la manipulación del módulo se lea detenidamente el Manual Técnico, para tener un enfoque claro del funcionamiento.The present titling work has the objective of developing an automatic colimetry system with artificial vision based on the subtractive model Cian, Magenta, Yellow, Black (CMYK). The system was implemented by designing the structure in computer-aided drawing (CAD) software, and then materials were acquired. The assembly of the structure was carried out to carry out the programming in a programmable logic controller (PLC), while the control system was realized in a human machine interface (HIM) by means of the software Lookout and the part of the acquisition of the color it done in the MATLAB programming software. After the tests, it was observed that the percentage of error of the values calculated by the implemented system with respect to the theoretical values of CMYK colors was approximately 3.8%; which corroborates the efficiency of the machine. The colimetry system allows assigning a percentage value for the four-color inks and making the blends to obtain a color captured by the cell phone camera. Students are advised that prior to manipulating the module, the Technical Manual should be carefully read, in order to have a clear focus on the operation

Aportaciones al conocimiento del Calcolítico de la Cuenca Media del Guadiana: la comarca de Zafra (Badajoz)

Las prospecciones arqueológicas desarrolladas en la comarca de Zafra han puesto a la luz la existencia de una serie de yacimientos pertenecientes a diversos períodos prehistóricos e históricos. Entre los primeros destaca un grupo de varios asentamientos de la Edad del Cobre que constituyen un muestreo significativo de este período. A pesar de tratarse de recogidas superficiales o, en el mejor de los casos, de elementos desconectados de sus primitivos contextos, la cantidad y calidad de muestras permite avanzar algunas hipótesis de cara a la cronología de estos enclaves y a la dinámica del poblamiento eneolítico de esta comarca bajoextremeña. El estudio del Calcolítico en esta zona viene así a sumarse a los ya efectuados para las comarcas de Llerena y Mérida (este último dentro del conjunto de la Cuenca Media del Guadiana) como una pieza más dentro del amplio espectro del poblamiento calcolítico suroccidental, en el convencimiento de que es el conocimiento profundo del registro arqueológico el único vehículo válido para establecer criterios firmes en el debate científico que propicia la construcción de la Prehistoria.Archaeological prospections developed in the region of Zafra have highlighted the existence of a series of sites belonging to various prehistoric and historical periods. Among the first, there is a group of several settlements from the Copper Age that constitute a significant sample of this period. In spite of being superficial collections or, in the best of cases, elements disconnected from their primitive contexts, the quantity and quality of samples allow us to advance some hypotheses regarding the chronology of these enclaves and the dynamics of the eneolithic population of this region. The study of the Chalcolithic in this area is thus added to those already made for the counties of Llerena and Mérida (the latter within the middle basin of the Guadiana) as one more piece within the broad spectrum of the southwest calcolithic settlement, in the conviction that it is the deep knowledge of the archaeological record the only valid vehicle to establish firm criteria in the scientific debate that favors the construction of Prehistory.peerReviewe

Methodological Approach in the Development of Specific Games in Elite Soccer

In games such as soccer, where the stability and the possibility of replicating game situations are complex, teams and players continually deal with a highly unstable cooperative and non-cooperative environment. Thus, synchronized cooperation among players during training sessions is a fundamental factor, which many times contributes to a team’s success. In this context, there are some specific drills that attempt to challenge and create meaningful contexts in order to simulate match situations as closely as possible. Small-sided games are play-sport situations in which all elements of the game interact together in a flexible manner. However, there are a variety of small-sided games in elite soccer, such as possession games and positional games, which may present specific characteristics and stimulate different physical-physiological demands. An adequate selection and implementation of these games may help coaches to promote positive adaptations and performance improvements. Thus, this chapter provides practical tips to modulate the physical-physiological responses and technical-tactical requirements of the players using a variety of game formats during soccer training sessions

Selective Detection of Aeromonas Spp. By a Fluorescent Probe Based on the Siderophore Amonabactin

[Abstract] Amonabactins, the siderophores produced by some pathogenic bacteria belonging to Aeromonas genus, can be used for the preparation of conjugates to be imported into the cell using their specific transport machinery. Herein, we report the design and synthesis of a new amonabactin-based fluorescent probe by conjugation of the appropriate amonabactin analogue to sulforhodamine B (AMB-SRB) using a thiol-maleimide click reaction. Growth promotion assays and fluorescence microscopy studies demonstrated that the AMB-SRB fluorescent probe was able to label the fish pathogenic bacterium A. salmonicida subsp. salmonicida through its outer membrane transport (OMT) protein FstC. The labelling of other Aeromonas species, such as the human pathogen A. hydrophila, indicates that this probe can be a very useful molecular tool for studying the amonabactin-dependent iron uptake mechanism. Furthermore, the selective labelling of A. salmonicida and other Aeromonas species in presence of other fish pathogenic bacteria, suggest the potential application of this probe for detection of Aeromonas in water and other fish farming samples through fluorescence assays.This work was supported by grants RTI2018-093634-B-C21/C22 from the State Agency for Research (AEI) of Spain, cofunded by the FEDER Programme from the European Union (MCIN/AEI/10.13039/501100011033/FEDER). M.B. was supported by grant PID2019-103891RJ-100 from MCIN/AEI (Spain). Work in University of Santiago de Compostela and University of A Coruña was also supported by grants GRC2018/018 and GRC2018/039, respectively, from Xunta de Galicia and BLUEBIOLAB (0474_BLUEBIOLAB_1_E), Programme INTERREG V A of Spain-Portugal (POCTEP). D.R-V. thanks Xunta de Galicia (Spain) for a predoctoral fellowship. J.C.-S. thanks to the FPU National Program (FPU16/02060) of the Spanish Ministry of Science, Innovation and Universities for a predoctoral fellowshipXunta de Galicia; GRC2018/018Xunta de Galicia; GRC2018/03