Process Intensification at the expression system level for the production of 1-phosphate aldolase in antibiotic-free E. coli fed-batch cultures

In order to successfully design expression systems for industrial biotechnology and biopharmaceutical applications; plasmid stability, capacity to efficiently synthesize the desired product and the use of selection markers that are acceptable to regulatory bodies are of utmost importance. In this work we demonstrate the application of a set of engineered strains – with different features namely, antibiotic vs auxotrophy marker; two-plasmids vs single plasmid expression system; expression levels of the repressor protein (LacI) and the auxotrophic marker (glyA) – in high cell density cultures to evaluate their suitability to be used in bioprocess conditions that resemble industrial production. Results revealed that the first generation of engineered strain showed a 50 % reduction in fuculose-1-phosphate aldolase (FucA) production compared to the reference system (165 ± 13 mg FucA·g-1 DCW and 806 ± 12 AU·g-1 DCW) which is commercially available from QIAGEN and uses an antibiotic selection marker. The over-transcription of the glyA gene was found to be a major factor responsible for the metabolic burden leading to the decrease in FucA yield. The second- and third-generation of E. coli strains presented an increase in FucA production, being 202 ± 18 mg-1 FucA·g-1 DCW and 1176 ± 19 AU·g-1 DCW, and 1322 ± 19 AU·g-1 DCW and 245 ± 13 mg-1 FucA·g-1 DCW, respectively. Both strains presented a fitness improvement after tuning the expression levels of the glyA gene and the deletion of the ampicillin resistance gene (bla) from the plasmid were carried out, respectively. The third-generation expression system is antibiotic-free, autotrophy-selection based and single-plasmid and is capable to produce FucA at similar levels compared to the original commercial expression system. Hence, our expression system possesses advantageous features compared to the commercial one and proved to have the potential to become an attractive platform for the production of recombinant proteins in a wide range of industrial biotechnology applications

Opening the black box of energy throughputs in farm systems: A decomposition analysis between the energy returns to external inputs, internal biomass reuses and total inputs consumed (the Vallès County, Catalonia, c.1860 and 1999)

We present an energy analysis of past and present farm systems aimed to contribute to their sustainability assessment. Looking at agroecosystems as a set of energy loops between nature and society, and adopting a farm-operator standpoint at landscape level to set the system boundaries, enthalpy values of energy carriers are accounted for net Final Produce going outside as well as for Biomass Reused cycling inside, and External Inputs are accounted using embodied values. Human Labour is accounted for the fraction of the energy intake of labouring people devoted to perform farm work, considering the local or external origin of their food basket. In this approach the proportion of internal Biomass Reused becomes a hallmark of organic farm systems that tend to save External Inputs, whereas industrial farming and livestock breeding in feedlots tend to get rid of reuses replacing them with inputs coming from outside. Hence, decomposing the internal or external energy throughputs may bring to light their contrasting sociometabolic profiles. A Catalan case study in 1860 and 1990 is used as a test bench to show how revealing this decomposing analysis may be to plot the energy profiles of farm systems and their possible improvement pathways

Widening the analysis of Energy Return on Investment (EROI) in agro-ecosystems: Socio-ecological transitions to industrialized farm systems (the Vallès County, Catalonia, c.1860 and 1999)

Energy balances of farm systems have overlooked the role of energy flows that remain within agro-ecosystems. Yet, such internal flows fulfil important socio-ecological functions, including maintenance of farmers themselves and agro-ecosystem structures. Farming can either give rise to complex landscapes that favour associated biodiversity, or the opposite. This variability can be understood by assessing several types of Energy Returns on Investment (EROI). Applying these measures to a farm system in Catalonia, Spain in 1860 and in 1999, reveals the expected decrease in the ratio of final energy output to total and external inputs. The transition from solar-based to a fossil fuel based agro-ecosystem was further accompanied by an increase in the ratio of final energy output to biomass reused, as well as an absolute increase of Unharvested Phytomass grown in derelict forestland. The study reveals an apparent link between reuse of biomass and the decrease of landscape heterogeneity along with its associated biodiversity

Novel method for rapid monitoring of OPFRs by LLE and GC-MS as a tool for assessing biodegradation: validation and applicability

Organophosphate flame retardants (OPFRs) are high-production volume chemicals widely present in environmental compartments. The presence of water-soluble OPFRs (tri-n-butyl phosphate (TnBP), tris(2-butoxyethyl) phosphate (TBEP), tris(2-chloroethyl) phosphate (TCEP), tris(2-chloroisopropyl) phosphate (TCPP), and triethyl phosphate (TEP)) in water compartments evidences the struggle of conventional wastewater treatment plants (WWTPs) to effectively eliminate these toxic compounds. This study reports for the first time the use of white-rot fungi as a promising alternative for the removal of these OPFRs. To accomplish this, a simple and cost-efficient quantification method for rapid monitoring of these contaminants' concentrations by GC-MS while accounting for matrix effects was developed. The method proved to be valid and reliable for all the tested parameters. Sample stability was examined under various storage conditions, showing the original samples to be stable after 60 days of freezing, while post-extraction storage techniques were also effective. Finally, a screening of fungal degraders while assessing the influence of the glucose regime on OPFR removal was performed. Longer chain organophosphate flame retardants, TBP and TBEP, could be easily and completely removed by the fungus Ganoderma lucidum after only 4 days. This fungus also stood out as the sole organism capable of partially degrading TCEP (35% removal). The other chlorinated compound, TCPP, was more easily degraded and 70% of its main isomer was removed by T. versicolor. However, chlorinated compounds were only partially degraded under nutrient-limiting conditions. TEP was either not degraded or poorly degraded, and it is likely that it is a transformation product from another OPFR's degradation. These results suggest that degradation of chlorinated compounds is dependent on the concentration of the main carbon source and that more polar OPFRs are less susceptible to degradation, given that they are less accessible to radical removal by fungi. Overall, the findings of the present study pave the way for further planned research and a potential application for the degradation of these contaminants in real wastewaters.Open Access Funding provided by Universitat Autonoma de Barcelona. This work has been supported by project PID2019-103989RB-100 financed by MCIN/ AEI /https://doi.org/10.13039/501100011033. This work has been partially supported by the Generalitat de Catalunya (Consolidated Research Group 2021-SGR-01008). Diana Losantos received support from MCIN predoctoral research grant ref PRE2020-095902.Peer reviewe

Digital image analysis of the human bite mark

SIGLEAvailable from British Library Document Supply Centre-DSC:DXN010688 / BLDSC - British Library Document Supply CentreGBUnited Kingdo

Cloning, expression, and one-step purification/immobilization of two carbohydrate-binding module-tagged alcohol dehydrogenases

The feasibility of biochemical transformation processes is usually greatly dependent on biocatalysts cost. Therefore, immobilizing and reusing biocatalysts is an approach to be considered to bring biotransformations closer to industrial feasibility, since it does not only allow to reuse enzymes but can also improve their stability towards several reaction conditions. Carbohydrate-Binding Modules (CBM) are well-described domains involved in substrate binding which have been already used as purification tags.This project has received funding from the European Union’s Horizon 2020 Research and Innovation programme under Grant Agreement n°761042 (BIOCON-CO2). This output reflects the views only of the author and the European Comission cannot be held responsible for any use which may be made of the information contained therein. The research group is recognized by Generalitat de Catalunya as 2017 SGR 1462.Peer reviewe

Fungal treatment for the removal of endocrine disrupting compounds from reverse osmosis concentrate: Identification and monitoring of transformation products of benzotriazoles

The removal of 27 endocrine-disrupting compounds and related compounds (suspect effect) from a reverse osmosis concentrate using an alternative decontamination method based on a fungal treatment involving Trametes versicolor was assessed. In addition to chemical analysis, the toxicity of the treated water during the treatment was monitored using a bioluminescence inhibition test and estrogenic and anti-estrogenic tests. The compounds 1H-benzotriazole (BTZ) and two tolyltriazoles (TTZs), 4-methyl-1H-benzotriazole (4-MBTZ) and 5-methyl-1H-benzotriazole (5-MBTZ), were present in the reverse osmosis concentrate at the highest concentrations (7.4 and 12.8 μg L −1 , respectively) and were partially removed by the fungal treatment under sterile conditions (58% for BTZ and 92% for TTZs) and non-sterile conditions, although to lesser extents (32% for BTZ and 50% for TTZs). Individual biotransformation studies of BTZ and the TTZs by T. versicolor in a synthetic medium and further analysis via on-line turbulent flow chromatography coupled to an HRMS-Orbitrap allowed the tentative identification of the transformation products (TPs). Six TPs were postulated for BTZ, two TPs were postulated for 4-MBTZ, and four TPs were postulated for 5-MBTZ. Most of these TPs are suggested to have been generated by conjugation with some sugars and via the methylation of the triazole group. Only TP 148 A, postulated to be derived from the biotransformation of BTZ, was observed in the effluent of the bioreactor treating the reverse osmosis concentrateThis work has been supported by the Spanish Ministry of Economy, Industry and Competitiveness (project CTQ2010-21776-C02 and CTM2013-48545-C2), co-financed by the European Union through the European Regional Development Fund (ERDF) and supported by the Generalitat de Catalunya (Consolidated Research Groups: Catalan Institute for water Research 2014 SGR 291 and 2014 SGR 476

Degradation of pharmaceuticals from membrane biological reactor sludge with Trametes versicolor

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Políticas públicas y equilibrio territorial en el Estado

Obra impulsada por el Institut d’Estudis Autonòmics de la Generalitat de Cataluña y la Fundación BBVA cuyo objetivo es analizar la incidencia territorial de la actuación del sector público español. Una parte importante del libro se centra en el análisis de los patrones territoriales de gasto e ingresos tributarios de los distintos niveles de la administración pública. El estudio de las políticas de gasto indica que las Comunidades Autónomas han hecho un uso efectivo de su autonomía y han exhibido una notable diferenciación en sus políticas de gasto para adecuar sus ofertas de bienes y servicios a las heterogéneas necesidades y preferencias de sus ciudadanos. Los autores de este estudio aprecian una reducción progresiva de las diferencias interregionales en la financiación básica per cápita de las CC.AA. de régimen común, aunque todavía queda un trecho significativo por recorrer. El diseño del sistema de financiación autonómica resulta también, sin perjuicio de sus fortalezas, claramente mejorable.Tras el análisis de las políticas presupuestarias realizado en los primeros capítulos, la obra profundiza en el estudio de la incidencia de estas políticas sobre la distribución territorial de la renta y la actividad económica. De su lectura se concluye que la actuación del sector público ha contribuido de manera muy notable a la reducción de las disparidades de renta per cápita entre regiones, eliminando aproximadamente una tercera parte de las diferencias de renta per capita. Este importante efecto redistributivo proviene en exclusiva de la recaudación tributaria.El libro contiene también un análisis de la convergencia regional y sus determinantes, con especial atención al papel de la inversión pública en infraestructuras, que ha sido claramente convergente durante el período autonómico. Por último, la obra ofrece un análisis tentativo de la distribución territorial de la localización de la actividad económica, a partir del estudio de la ubicación de las sedes sociales de las grandes empresas.La obra facilita a los investigadores, responsables políticos y a la opinión pública especializada un amplio balance de las primeras dos décadas de funcionamiento del sistema autonómico en España.Finnancial support of the BBVA FoundationPeer reviewe