Production of secondary metabolites in cell suspension cultures of Ocimum sanctum L.

Dissertação de mestrado em Plant Molecular Biology, Biotechnology and Bioentrepreneurship

Search for the associated production of the Higgs boson with a top quark pair in multilepton final states with the ATLAS detector

A search for the associated production of the Higgs boson with a top quark pair is performed in multilepton final states using 20.3 fb−1 of proton-proton collision data recorded by the ATLAS experiment at s√=8 TeV at the Large Hadron Collider. Five final states, targeting the decays H→WW∗, ττ, and ZZ∗, are examined for the presence of the Standard Model (SM) Higgs boson: two same-charge light leptons (e or μ) without a hadronically decaying τ lepton; three light leptons; two same-charge light leptons with a hadronically decaying τ lepton; four light leptons; and one light lepton and two hadronically decaying τ leptons. No significant excess of events is observed above the background expectation. The best fit for the tt¯H production cross section, assuming a Higgs boson mass of 125 GeV, is 2.1+1.4−1.2 times the SM expectation, and the observed (expected) upper limit at the 95% confidence level is 4.7 (2.4) times the SM rate. The p-value for compatibility with the background-only hypothesis is 1.8σ; the expectation in the presence of a Standard Model signal is 0.9σ.

ATLAS Run 1 searches for direct pair production of third-generation squarks at the Large Hadron Collider

This paper reviews and extends searches for the direct pair production of the scalar supersymmetric partners of the top and bottom quarks in proton--proton collisions collected by the ATLAS collaboration during the LHC Run 1. Most of the analyses use 20 fb−1 of collisions at a centre-of-mass energy of s√=8 TeV, although in some case an additional 4.7 fb−1 of collision data at s√=7 TeV are used. New analyses are introduced to improve the sensitivity to specific regions of the model parameter space. Since no evidence of third-generation squarks is found, exclusion limits are derived by combining several analyses and are presented in both a simplified model framework, assuming simple decay chains, as well as within the context of more elaborate phenomenological supersymmetric models.

Parameter identification of the fermentative production of fructo-oligosaccharides by Aureobasidium pullulans

In this study, a mathematical model for the production of Fructo-oligosaccharides (FOS) by Aureobasidium pullulans is developed. This model contains a relatively large set of unknown parameters, and the identification problem is analyzed using simulation data, as well as experimental data. Batch experiments were not sufficiently informative to uniquely estimate all the unknown parameters, thus, additional experiments have to be achieved in fed-batch mode to supplement the missing information. © 2015 IEEE.

Production of elastic Chitosan/Chondroitin sulphate multilayer films for tissue regeneration

Membrane-like scaffolds are suitable to induce regeneration in many and different anatomic sites, such as periodontal membrane, skin, liver and cardiac tissues. In some circumstances, the films should adapt to geometrical changes of the attached tissues, such as in cardiac or blood vessel tissue engineering applications. In this context, we developed stretchable two-dimensional multilayer constructs through the assembling of two natural-based polyelectrolytes, chitosan (CHT) and chondroitin sulphate (CS), using the layer-by-layer methodology. The morphology, topography and the transparency of the films were evaluated. The in- fluence of genipin, a natural-derived cross-linker agent, was also investigated in the control of the mechanical properties of the CHT/CS films. The water uptake ability can be tailored by changing the cross-linker concentration, which influenced the young modulus and ultimate tensile strength. The maximum extension tends to decrease with the increase of genipin concentration, compromising the elastic properties of CHT/CS films: nevertheless using lower cross-linker contents, the ultimate tensile stress is similar to the films not cross-linked but exhibiting a significant higher modulus. The in vitro biological assays showed better L929 cell adhesion and proliferation when using the crosslinked membranes and confirmed the non-cytotoxicity of the CHT/CS films. The developed free-standing biomimetic multilayer could be designed to fulfill specific therapeutic requirements by tuning properties such as swelling, mechanical and biological performances.

Synthetic biology approaches to engineer new pathways for the production of plant secondary metabolites

Secondary metabolites from plants are important sources of high-value chemicals, many of them being pharmacologically active. These metabolites are commonly isolated through inefficient extractions from natural biological sources and are often difficult to synthesize chemically. Therefore, their production using engineered organisms has lately attracted an increased attention. Curcuminoids, an example of such metabolites, are produced in Curcuma longa and exhibit anti-cancer and anti-inflammatory activities. Herein we report the construction of an artificial biosynthetic pathway for the curcuminoids production in Escherichia coli. Different 4-coumaroyl-CoA ligases (4CL) and polyketide synthases (diketide-CoA synthase (DCS), curcumin synthase (CURS) and curcuminoid synthase) were tested. The highest curcumin production (70 mg/L) was obtained by feeding ferulic acid and with the Arabidopsis thaliana 4CL1 and C. longa DCS and CURS enzymes. Other curcuminoids (bisdemethoxy- and demethoxycurcumin) were also produced by feeding coumaric acid or a mixture of coumaric and ferulic acids, respectively. Curcuminoids, including curcumin, were also produced from tyrosine through the caffeic acid pathway. To produce caffeic acid, tyrosine ammonia lyase and 4-coumarate 3-hydroxylase were used. Caffeoyl-CoA O-methyltransferase was used to convert caffeoyl-CoA to feruloyl-CoA. This pathway represents an improvement of the curcuminoids heterologous production. The construction of this pathway in another model organism is being considered, as well as the introduction of alternative enzymes.

Production of medium-chain fatty acids and higher alcohols by a synthetic co-culture grown on carbon monoxide or syngas

Synthesis gas, a mixture of CO, H2, and CO2, is a promising renewable feedstock for bio-based production of organic chemicals. Production of medium-chain fatty acids can be performed via chain elongation, utilizing acetate and ethanol as main substrates. Acetate and ethanol are main products of syngas fermentation by acetogens. Therefore, syngas can be indirectly used as a substrate for the chain elongation process.

Microbial production of potent phenolic-antioxidants through solid state fermentation

The agroindustrial residues including plant tissues rich in polyphenols were explored for microbial production of potent phenolics under solid state fermentation processes. The fungal strains capable of hydrolyzing tannin-rich materials were isolated from Mexican semidesert zones. These microorganisms have been employed to release potent phenolic antioxidants during the solid state fermentation of different materials (pomegranate peels, pecan nut shells, creosote bush and tar bush). This chapter includes the critical parameters for antioxidants production from selective microbes. Technical aspects of the microbial fermentation of antioxidants have also been discussed.

Valorization of agro-industrial wastes towards the production of rhamnolipids

In this work, oil mill wastewater (OMW), a residue generated during olive oil extraction, was evaluated as an inducer of rhamnolipid production. Using a medium containing as sole ingredients corn steep liquor (10%, v/v), sugarcane molasses (10%, w/v) and OMW (25%, v/v), Pseudomonas aeruginosa #112 produced 4.5 and 5.1 g of rhamnolipid per liter in flasks and reactor, respectively, with critical micelle concentrations as low as 13 mg/l. Furthermore, in the medium supplemented with OMW, a higher proportion of more hydrophobic rhamnolipid congeners was observed comparing with the same medium without OMW. OMW is a hazardous waste which disposal represents a serious environmental problem; therefore, its valorization as a substrate for the production of added-value compounds such as rhamnolipids is of great interest. This is the first report of rhamnolipid production using a mixture of these three agro-industrial by-products, which can be useful for the sustainable production of rhamnolipids.

Production of whey protein-based aggregates under ohmic heating

Formation of whey protein isolate protein aggregates under the influence of moderate electric fields upon ohmic heating (OH) has been monitored through evaluation of molecular protein unfolding, loss of its solubility, and aggregation. To shed more light on the microstructure of the protein aggregates produced by OH, samples were assayed by transmission electron microscopy (TEM). Results show that during early steps of an OH thermal treatment, aggregation of whey proteins can be reduced with a concomitant reduction of the heating chargeby reducing the come-up time (CUT) needed to reach a target temperatureand increase of the electric field applied (from 6 to 12 V cm1). Exposure of reactive free thiol groups involved in molecular unfolding of -lactoglobulin (-lg) can be reduced from 10 to 20 %, when a CUT of 10 s is combined with an electric field of 12 V cm1. Kinetic and multivariate analysis evidenced that the presence of an electric field during heating contributes to a change in the amplitude of aggregation, as well as in the shape of the produced aggregates. TEM discloses the appearance of small fibrillar aggregates upon the influence of OH, which have recognized potential in the functionalization of food protein networks. This study demonstrated that OH technology can be used to tailor denaturation and aggregation behavior of whey proteins due to the presence of a constant electric field together with the ability to provide a very fast heating, thus overcoming heat transfer limitations that naturally occur during conventional thermal treatments.

Characterization of late antique and early medieval pottery production of the city of Braga and its territory

The fundamental goal we set ourselves when developing this study is to try to characterize, both technically and formally, ceramics made in the city of Braga and its territory from the initial moments of the Late Antiquity to the Middle Ages. Thus, we will focus on analyzing some own productions that appear attached to the phases of late antique occupation —ceramics of red engobes and late gray—, as well as in the early medieval containers identified in different archaeological interventions practiced in the Braga environment. Concretely, we will analyze the material from various excavations conducted recently at the Theatre in the solar number 20/28 and 36/56 from the Afonso Henriques Street and the former District Hostel as well as the church of São Martinho de Dume.

Production of biohydrogen from dark fermentation of organic wastes

Dissertação de mestrado em Bioengenharia

Exploitation of Ashbya gossypii for the production of high-value products from glycerol feedstocks

Dissertação de mestrado em Bioengenharia

Effect of arginine vasopressin on the canine epicardial coronary artery: experiments on V1-receptor-mediated production of nitric oxide

OBJECTIVE: To determine whether arginine vasopressin releases endothelium-derived nitric oxide (EDNO) from the epicardial coronary artery. METHODS: We studied segments of canine left circumflex coronary arteries suspended in organ chambers to measure isometric force. The coronary artery segments were contracted with prostaglandin F2alpha (2 x 10-6M) and exposed to a unique, strong arginine vasopressin concentration (10-6M) or titrated concentrations (10-9 a 10-5 M). RESULTS: The unique dose of arginine vasopressin concentration (10-6M) induced transient, but significant (p<0.05), relaxation in arterial segments with endothelium, and an increase, not significant, in tension in arteries without endothelium. Endothelium-dependent relaxation to arginine vasopressin was inhibited by Ng-monomethyl-L-arginine (L-NMMA, 10-5M) or N G-nitro-L-arginine (L-NOARG) (10-4M), 2 inhibitors of nitric oxide synthesis from L-arginine. Exogenous L-arginine (10-4M), but not D-arginine (10-4M), reversed the inhibitory effect of L-NMMA on vasopressin-mediated vasorelaxation. Endothelium dependent relaxation to vasopressin was also reversibly inhibited by the vasopressin V1-receptor blocker d(CH2)5Try(Me) arginine vasopressin (10-6M) (n=6, P<0.05). CONCLUSION: Vasopressin acts through V1 endothelial receptors to stimulate nitric oxide release from L-arginine.

Producción de ácido Itacónico por Aspergillus terreus y de Ácido Hialurónico por Streptococcus equi subsp. equi en Biorreactores de Tanque Agitado. Production of Itaconic Acid by Aspergillus terreus and of Hyaluronic Acid by Sreptococcus equi subsp. equi in Stirred Tank Bioreactors.

El proyecto aborda el problema general de establecer una metodología para el cambio de escala de la producción de metabolitos en biorreactores de tanque agitado.En el desarrollo de procesos de producción de metabolitos a partir de microorganismos, el cambio de escala es particularmente complejo, dado que los microorganismos experimentan un continuo cambio en sus rutas metabólicas durante el período de producción. Esto hace que en el proceso del cambio de escala, las mayores dificultades se encuentren en el desarrollo del inóculo y problemas ocasionados por modificaciones en las características de transferencia de calor, masa y momento.Dentro de este contexto general se definen dos objetivos específicos. Estos son: el estudio de la producción de ácido itacónico por Aspergillus terreus y el de la producción de ácido hialurónico por Streptococcus equi. subsp. equi, con la finalidad de desarrollar una metodología de trabajo experimental y teórica que permita sistematizar el estudio del factibilidad técnico-económica de plantas de producción, vinculando la investigación del procesos a escala de laboratorio con la producción a mayor escala.La hipótesis de trabajo es que el estudio de la producción de Aspergillus terreus y de Streptococcus equi en un biorreactor de tanque agitado a escala de laboratorio permitirá establecer los parámetros que contribuirán a realizar el cambio de escala de su producción y esto será verificado experimentalmente.Los trabajos se realizarán utilizando un biorreactor a escala de laboratorio especialmente diseñado para este tipo de trabajo. Los resultados experimentales se interpretarán con técnicas estadísticas y matemáticas de diferente complejidad a efectos de establecer los criterios de cambio de escala y luego se realizarán experiencias en un biorreactor piloto con el objeto de verificar la metodología seleccionada.El desarrollo del proyecto permitirá:1.- obtener información técnica útil sobre la producción de ácido itacónico, el que tiene importantes aplicaciones en la industria del plástico. La producción por medio del Aspergillus terreus MJL05 se realizará utilizando glicerol como fuente de carbono, el que constituye el principal subproducto en los procesos de manufactura de biodiesel. De este modo se podrá analizar la factibilidad técnica de una ruta alternativa para emplear este subproducto.2.- obtener información técnica útil sobre la producción de ácido hialurónico, biopolímero de alto valor agregado con importantes aplicaciones en medicina, y contribuir así a realizar el cambio de escala de su producción.