Bioethanol production by Saccharomyces cerevisiae, Pichia stipitis and Zymomonas mobilis from delignified coconut fibre mature and lignin extraction according to biorefinery concept

In search to increase the offer of liquid, clean, renewable and sustainable energy in the world energy matrix, the use of lignocellulosic materials (LCMs) for bioethanol production arises as a valuable alternative. The objective of this work was to analyze and compare the performance of Saccharomyces cerevisiae, Pichia stipitis and Zymomonas mobilis in the production of bioethanol from coconut fibre mature (CFM) using different strategies: simultaneous saccharification and fermentation (SSF) and semi-simultaneous saccharification and fermentation (SSSF). The CFM was pretreated by hydrothermal pretreatment catalyzed with sodium hydroxide (HPCSH). The pretreated CFM was characterized by X-ray diffractometry and SEM, and the lignin recovered in the liquid phase by FTIR and TGA. After the HPCSH pretreatment (2.5% (v/v) sodium hydroxide at 180 °C for 30 min), the cellulose content was 56.44%, while the hemicellulose and lignin were reduced 69.04% and 89.13%, respectively. Following pretreatment, the obtained cellulosic fraction was submitted to SSF and SSSF. Pichia stipitis allowed for the highest ethanol yield 90.18% in SSSF, 91.17% and 91.03% were obtained with Saccharomyces cerevisiae and Zymomonas mobilis, respectively. It may be concluded that the selection of the most efficient microorganism for the obtention of high bioethanol production yields from cellulose pretreated by HPCSH depends on the operational strategy used and this pretreatment is an interesting alternative for add value of coconut fibre mature compounds (lignin, phenolics) being in accordance with the biorefinery concept.

Improvement of in silico strain engineering methods in Saccharomyces cerevisiae

PhD thesis in Bioengineering

In silico constraint-based strain optimization methods: the quest for optimal cell factories

Shifting from chemical to biotechnological processes is one of the cornerstones of 21st century industry. The production of a great range of chemicals via biotechnological means is a key challenge on the way toward a bio-based economy. However, this shift is occurring at a pace slower than initially expected. The development of efficient cell factories that allow for competitive production yields is of paramount importance for this leap to happen. Constraint-based models of metabolism, together with in silico strain design algorithms, promise to reveal insights into the best genetic design strategies, a step further toward achieving that goal. In this work, a thorough analysis of the main in silico constraint-based strain design strategies and algorithms is presented, their application in real-world case studies is analyzed, and a path for the future is discussed.

Clivagem e caracterização de frutalina recombinante produzida em Escherichia coli

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Engenharia Clínica)

Centrality and rapidity dependence of inclusive jet production in sNN=5.02 TeV proton-lead collisions with the ATLAS detector

Measurements of the centrality and rapidity dependence of inclusive jet production in sNN−−−√=5.02 TeV proton--lead (p+Pb) collisions and the jet cross-section in s√=2.76 TeV proton--proton collisions are presented. These quantities are measured in datasets corresponding to an integrated luminosity of 27.8 nb−1 and 4.0 pb−1, respectively, recorded with the ATLAS detector at the Large Hadron Collider in 2013. The p+Pb collision centrality was characterised using the total transverse energy measured in the pseudorapidity interval −4.9<η<−3.2 in the direction of the lead beam. Results are presented for the double-differential per-collision yields as a function of jet rapidity and transverse momentum (pT) for minimum-bias and centrality-selected p+Pb collisions, and are compared to the jet rate from the geometric expectation. The total jet yield in minimum-bias events is slightly enhanced above the expectation in a pT-dependent manner but is consistent with the expectation within uncertainties. The ratios of jet spectra from different centrality selections show a strong modification of jet production at all pT at forward rapidities and for large pT at mid-rapidity, which manifests as a suppression of the jet yield in central events and an enhancement in peripheral events. These effects imply that the factorisation between hard and soft processes is violated at an unexpected level in proton--nucleus collisions. Furthermore, the modifications at forward rapidities are found to be a function of the total jet energy only, implying that the violations may have a simple dependence on the hard parton--parton kinematics.

Measurement of the forward-backward asymmetry of electron and muon pair-production in pp collisions at s√ = 7 TeV with the ATLAS detector

This paper presents measurements from the ATLAS experiment of the forward-backward asymmetry in the reaction pp→Z/γ∗→l+l−, with l being electrons or muons, and the extraction of the effective weak mixing angle. The results are based on the full set of data collected in 2011 in pp collisions at the LHC at s√ = 7 TeV, corresponding to an integrated luminosity of 4.8 fb−1. The measured asymmetry values are found to be in agreement with the corresponding Standard Model predictions. The combination of the muon and electron channels yields a value of the effective weak mixing angle of 0.2308±0.0005(stat.)±0.0006(syst.)±0.0009(PDF), where the first uncertainty corresponds to data statistics, the second to systematic effects and the third to knowledge of the parton density functions. This result agrees with the current world average from the Particle Data Group fit.

Measurements of the Total and Differential Higgs Boson Production Cross Sections Combining the H→γγ and H→ZZ∗→4ℓ Decay Channels at s√=8 TeV with the ATLAS Detector

Measurements of the total and differential cross sections of Higgs boson production are performed using 20.3 fb−1 of pp collisions produced by the Large Hadron Collider at a center-of-mass energy of s√=8 TeV and recorded by the ATLAS detector. Cross sections are obtained from measured H→γγ and H→ZZ∗→4ℓ event yields, which are combined accounting for detector efficiencies, fiducial acceptances and branching fractions. Differential cross sections are reported as a function of Higgs boson transverse momentum, Higgs boson rapidity, number of jets in the event, and transverse momentum of the leading jet. The total production cross section is determined to be σpp→H=33.0±5.3(stat)±1.6(sys)pb. The measurements are compared to state-of-the-art predictions.

Biosynthetic production of curcuminoids

Curcuminoids are natural phenylpropanoids from plants that have been reported as potential cancer-fighting drugs. Nevertheless, these compounds present a poor bioavailability. Cellular uptake is low and curcuminoids are quickly metabolized once inside the cell, requiring repetitive oral doses to achieve an effective concentration for therapeutic activity [1]. Herein, we report an engineered artificial pathway for the production of curcuminoids in Escherichia coli. Arabidopsis thaliana 4-coumaroyl-CoA ligase and Curcuma longa diketide-CoA synthase (DCS) and curcumin synthase (CURS1) were used and 188 µM (70 mg/L) of curcumin was obtained from ferulic acid [2]. Bisdemethoxycurcumin and demethoxycurcumin were also produced, but in lower concentrations, by feeding p-coumaric acid or a mixture of p-coumaric acid and ferulic acid, respectively. Additionally, curcuminoids were produced from tyrosine through the caffeic acid pathway. To produce caffeic acid, tyrosine ammonia lyase from Rhodotorula glutinis and 4-coumarate 3-hydroxylase from Saccharothrix espanaensis were used [3]. Caffeoyl-CoA 3-O-methyl-transferase from Medicago sativa was used to convert caffeoyl-CoA to feruloyl-CoA. Using caffeic acid, p-coumaric acid or tyrosine as a substrate, 3.9, 0.3, and 0.2 µM of curcumin were produced, respectively. This is the first report on the use of DCS and CURS1 in vivo to produce curcuminoids. In addition, curcumin, the most studied curcuminoid for therapeutic purposes and considered in many studies as the most potent and active, was produced by feeding tyrosine using a pathway involving caffeic acid. We anticipate that by using a tyrosine overproducing strain, curcumin can be produced in E. coli without the need of adding expensive precursors to the medium, thus decreasing the production cost. Therefore, this alternative pathway represents a step forward in the heterologous production of curcumin using E. coli. Aiming at greater production titers and yields, the construction of this pathway in another model organism such as Saccharomyces cerevisiae is being considered.

Cheese whey: a cost-effective alternative for hyaluronic acid production by Streptococcus zooepidemicus

This study focuses on the optimization of cheese whey formulated media for the production of hyaluronic acid HA by Streptococcus zooepidemicus. Culture media containing whey (W; 2.1 g/L) or whey hydrolysate (WH; 2.4 g/L) gave the highest HA productions. Both W and WH produced high yields on protein consumed, suggesting cheese whey is a good nitrogen source for S. zooepidemicus production of HA. Polysaccharide concentrations of 4.0 g/L and 3.2 g/L were produced in W and WH in a further scale-up to 5 L bioreactors, confirming the suitability of the low-cost nitrogen source. Cheese whey culture media provided high molecular weight (> 3000 kDa) HA products. This study revealed replacing the commercial peptone by the low-cost alternative could reduce HA production costs by up to a 70%compared to synthetic media.

Measurement of the t(t)over-barW and t(t)over-barZ production cross sections in pp collisions at root s=8 TeV with the ATLAS detector

The production cross sections of top-quark pairs in association with massive vector bosons have been measured using data from pp collisions at s√=8 TeV. The dataset corresponds to an integrated luminosity of 20.3 fb−1 collected by the ATLAS detector in 2012 at the LHC. Final states with two, three or four leptons are considered. A fit to the data considering the tt¯W and tt¯Z processes simultaneously yields a significance of 5.0σ (4.2σ) over the background-only hypothesis for tt¯W (tt¯Z) production. The measured cross sections are σtt¯W=369+100−91 fb and σtt¯Z=176+58−52 fb. The background-only hypothesis with neither tt¯W nor tt¯Z production is excluded at 7.1σ. All measurements are consistent with next-to-leading-order calculations for the tt¯W and tt¯Z processes.

Biotechnological production and application of fructooligosaccharides

Currently, prebiotics are all carbohydrates of relatively short chain length. An important group is the fructooligosaccharides, which are a special kind of prebiotics associated to their selective stimulation of the activity of certain groups of colonic bacteria that have a positive and beneficial effect on intestinal microbiota, reducing incidence of gastrointestinal infections, respiratory and also possessing a recognized bifidogenic effect. Traditionally, these prebiotic compounds have been obtained through extraction processes from some plants, as well as through enzymatic hydrolysis of sucrose. However, different fermentative methods have also been proposed for the production of fructooligosaccharides, such as solid-state fermentation utilizing various agroindustrial by-products. By optimizing the culture parameters, fructooligosaccharides yields and productivity can be improved. The use of immobilized enzymes and cells has also been proposed as being an effective and economic method for large-scale production of fructooligosaccharides. This paper is an overview on the results of recent studies on fructooligosacharides biosynthesis, physicochemical properties, sources, biotechnological production and applications.

Production of mexican brown macroalgae fucoidan and fucosidases under an integral green technology bioproceses by the biorefinery concept

Marine ecosystem can be considered a rather exploited source of natural substances with enormous bioactive potential. In Mexico macro-algae study remain forgotten for research and economic purposes besides the high amount of this resource along the west and east coast. For that reason the Bioferinery Group of the Autonomous University of Coahuila, have been studying the biorefinery concept in order to recover high value byproducts of Mexican brown macro-algae including polysaccharides and enzymes to be applied in food, pharmaceutical and energy industry. Brown macroalgae are an important source of fucoidan, alginate and laminarin which comprise a complex group of macromolecules with a wide range of important biological properties such as anticoagulant, antioxidant, antitumoral and antiviral and also as rich source of fermentable sugars for enzymes production. Additionally, specific enzymes able to degrade algae matrix (fucosidases, sulfatases, aliginases, etc) are important tools to establish structural characteristics and biological functions of these polysaccharides. The aims of the present work were the integral study of bioprocess for macroalgae biomass exploitation by the use of green technologies as hydrothermal extraction and solid state fermentation in order to produce polysaccharides and enzymes (fucoidan and fucoidan hydrolytic enzymes). This work comprises the use of the different bioprocess phases in order to produce high value products with lower time and wastes.

Exploring the use of HPFRC and GFRP grids for the production of manhole covers

Corrosion of the steel reinforced concrete elements is one of the common pathologies that limits the long-term performance of urban infrastructures. This problem causes the loss of structural serviceability by decreasing the concrete-steel bond strength and reducing the cross section of the reinforcements. The present study introduces a new system for developing free-corrosion resistance prefabricated manhole covers for applications in the aggressive environments, i.e. wastewater collector systems, sewer systems, stormwater systems, etc. Fibre reinforced cement composites were applied in this system in order to suppress the corrodible steel mesh and maintain the structural ductility as well. Application of fibre reinforced polymer (FRP) system is adopted as the additional solution for increasing the load carrying capacity of these elements without concerns about corrosion. The effectiveness of the applied strategy in developing the manhole covers in terms of load carrying capacity and failure mode is evaluated in this research. Furthermore, this paper discusses a FEM-based simulation, aiming to address the possibility of calibrating the constitutive model parameters related to fracture modes I and II.

Workplace ergonomics in lean production environments: A literature review

BACKGROUND: Lean Production Systems (LPS) have become very popular among manufacturing industries, services and large commercial areas. A LPS must develop and consider a set of work features to bring compatibility with workplace ergonomics, namely at a muscular, cognitive and emotional demands level. OBJECTIVE: Identify the most relevant impacts of the adoption of LPS from the ergonomics point of view and summarizes some possible drawbacks for workplace ergonomics due to a flawed application of the LPS. The impacts identified are focused in four dimensions: work pace, intensity and load; worker motivation, satisfaction and stress; autonomy and participation; and health outcome. This paper also discusses the influence that the work organization model has on workplace ergonomics and on the waste elimination previewed by LPS. METHODS: Literature review focused LPS and its impact on occupational ergonomics conditions, as well as on the Health and Safety of workers. The main focus of this research is on LPS implementations in industrial environments and mainly in manufacturing industry workplaces. This is followed by a discussion including the authors’ experience (and previous research). RESULTS: From the reviewed literature it seems that there is no consensus on how Lean principles affect the workplace ergonomics since most authors found positive (advantages) and negative (disadvantages) impacts. CONCLUSIONS: The negative impacts or disadvantages of LPS implementations reviewed may result from the misunderstanding of the Lean principles. Possibly, they also happen due to partial Lean implementations (when only one or two tools were implemented) that may be effective in a specific work context but not suitable to all possible situations as the principles of LPS should not lead, by definition, to any of the reported drawbacks in terms of workplace ergonomics.

Implementation of a lean six sigma project in a production line

Companies from the motorcycles components branch are dealing with a dynamic environment, resulting from the introduction of new products and the increase of market demand. This dynamic environment requires frequent changes in production lines and requires flexibility in the processes, which can cause reductions in the level of quality and productivity. This paper presents a Lean Six Sigma improvement project performed in a production line of the company's machining sector, in order to eliminate losses that cause low productivity, affecting the fulfillment of the production plan and customer satisfaction. The use of Lean methodology following the DMAIC stages allowed analyzing the factors that influence the line productivity loss. The major problems and causes that contribute to a reduction on productivity and that were identified in this study are the lack of standardization in the setup activities and the excessive stoppages for adjustment of the processes that caused an increase of defects. Control charts, Pareto analysis and cause-and-effect diagrams were used to analyze the problem. On the improvement stage, the changes were based on the reconfiguration of the line layout as well as the modernization of the process. Overall, the project justified an investment in new equipment, the defective product units were reduced by 84% and an increase of 29% of line capacity was noticed.