Síntese quimioenzimática do levetiracetam e análogos

Pós-graduação em Química - IQ

Effect of temperature on salt-salt aqueous biphasic systems: manifestations of upper critical solution temperature

Water-miscible ionic liquids (ILs) may be salted out using kosmotropic salts such as potassium phosphate (K3PO4) to form salt-salt aqueous biphasic systems (ABS). The effect of temperature on these systems has been studied using phase diagrams and it is observed that the degree of binodal shift decreases (requiring lower IL and kosmotropic salt concentrations) with the increase of temperature following the trend [C(4)mim]Cl > [C(4)py]Cl > [C(4)mmim] Cl > [N-4444]Cl. This trend can be correlated with the decreasing hydrogen bonding abilities of each salt. The phase behavior was also interpreted on the basis of critical solution temperature behavior of pure aqueous ionic liquid solutions. Additionally, the distribution of alcohols in these systems was studied as a function of temperature and it was found that the distribution ratios did not change with changes in temperature. The Gibbs energy of transfer of a methylene group in these systems and correlation to tie-line length was also determined. It was concluded that while the miscibility of alcohols increases in the ILs with increasing temperature, phase divergence in the aqueous biphasic system decreases, and thus these competing forces tend to cancel each other out for small polar molecules. A comparison is provided for the response to temperature in the currently studied salt-salt systems and analogous ABS formed by the addition of hydrophilic polymers to kosmotropic salts (polymer-salt) or other polymers (polymer-polymer).

Biomolecules extracted by ATPs: practical examples

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of the use of Aliquat 336 in the immobilization procedure in sol-gel of lipase from Bacillus sp ITP-001

Aliquat 336, a liquid hydrophobic material, was used at different concentrations (0.5-3.0%, w/v) as an additive in the preparation of encapsulated lipase from Bacillus sp. ITP-001 on sol-gel silica matrices using tetraethoxysilane (TEOS) as the precursor. The resulting hydrophobic matrices and immobilized lipases were characterized with regard to specific surface area (BET method), adsorption-desorption isotherms, pore volume (Vp) and size (dp) by nitrogen adsorption (BJH method) and scanning electron microscopy (SEM). The catalytic activities and the corresponding coupling yields were assayed in the hydrolysis of olive oil. In comparison with pure silica matrices, the immobilization process in the presence of Aliquat 336 decreased the values for specific surface area and increased the values for pore specific volume (Vp) and mean pore diameter (dp). This behavior may be related to the partial adsorption of the enzyme on the external surface of the hydrophobic matrix as indicated by scanning electron microscopy. Aliquat 336 concentrations in the range from 0.5 to 1.5% (w/v) provided immobilized derivatives with higher coupling yields and better substrate affinity. The highest coupling yield (Y-A = 71%) was obtained for the immobilized enzyme prepared in the presence of 1.5% Aliquat which gave the following morphological properties: specific surface area = 183 m(2)/g, pore specific volume (Vp) = 0.36 cc/g and mean pore diameter (dp)= 91 angstrom. (c) 2012 Elsevier B.V. All rights reserved.

First report on the kinetics of the uncatalyzed esterification of cellulose under homogeneous reaction conditions: a rationale for the effect of carboxylic acid anhydride chain-length on the degree of biopolymer substitution

The kinetics of the homogeneous acylation of microcrystalline cellulose, MCC, with carboxylic acid anhydrides with different acyl chain-length (Nc; ethanoic to hexanoic) in LiCl/N,N-dimethylacetamide have been studied by conductivity measurements from 65 to 85 A degrees C. We have employed cyclohexylmethanol, CHM, and trans-1,2-cyclohexanediol, CHD, as model compounds for the hydroxyl groups of the anhydroglucose unit of cellulose. The ratios of rate constants of acylation of primary (CHM; Prim-OH) and secondary (CHD; Sec-OH) groups have been employed, after correction, in order to split the overall rate constants of the reaction of MCC into contributions from the discrete OH groups. For the model compounds, we have found that k((Prim-OH))/k((Sec-OH)) > 1, akin to reactions of cellulose under heterogeneous conditions; this ratio increases as a function of increasing Nc. The overall, and partial rate constants of the acylation of MCC decrease from ethanoic- to butanoic-anhydride and then increase for pentanoic- and hexanoic anhydride, due to subtle changes in- and compensations of the enthalpy and entropy of activation.

Characterization of flexible DNA films

Polymer electrolytes (PEs) are currently the focus of much attention as potential electrolytes in electrochemical devices such as batteries, display devices and sensors. Deoxyribonucleic acid (DNA) as an important biological macromolecule has electric conducting electrochemical properties and unique three dimensional structures. With the goal of developing a new family of environmentally friendly multifunctional biohybrid materials displaying simultaneously high ionic conductivity we have produced in the present work, flexible films based on DNA, incorporating ionic liquids (ILs). Over the last decade ILs have been employed as a new media in electrochemistry and electroanalysis. The materials studied here have been characterized by means of Differential Scanning Calorimetry, Complex Impedance Spectroscopy and Cyclic Voltammetry. (C) 2012 Elsevier B.V. All rights reserved.

Enzyme Microheterogeneous Hydration and Stabilization in Supercritical Carbon Dioxide

Supercritical carbon dioxide is a promising green-chemistry solvent for many enzyme-catalyzed chemical reactions, yet the striking stability of some enzymes in such unconventional environments is not well understood. Here, we investigate the stabilization of the Candida antarctica Lipase B (CALB) in supercritical carbon dioxide-water biphasic systems using molecular dynamics simulations. The preservation of the enzyme structure and optimal activity depend on the presence of small amounts of water in the supercritical dispersing medium. When the protein is at least partially hydrated, water molecules bind to specific sites on the enzyme surface and prevent carbon dioxide from penetrating its catalytic core. Strikingly, water and supercritical carbon dioxide cover the protein surface quite heterogeneously. In the first solvation layer, the hydrophilic residues at the surface of the protein are able to pin down patches of water, whereas carbon dioxide solvates preferentially hydrophobic surface residues. In the outer solvation shells, water molecules tend to cluster predominantly on top of the larger water patches of the first solvation layer instead of spreading evenly around the remainder of the protein surface. For CALB, this exposes the substrate-binding region of the enzyme to carbon dioxide, possibly facilitating diffusion of nonpolar substrates into the catalytic funnel. Therefore, by means of microheterogeneous solvation, enhanced accessibility of hydrophobic substrates to the active site can be achieved, while preserving the functional structure of the enzyme. Our results provide a molecular picture on the nature of the stability of proteins in nonaqueous media.

Stereoselective synthesis of novel diels-alder adducts of p-substituted Methyl Thiocinnamates and Cyclopentadiene

This article describes the Diels-Alder reaction between methyl thiocinnamates, substituted at the para position by electron-donating and electron-withdrawing groups, with cyclopentadiene in the presence of catechol boron bromide (CBB) as a Lewis acid catalyst. The adduct configuration was confirmed by H-1 NMR coupling constants and single-crystal x-ray diffraction. Total endo stereoselectivity was observed in all reactions and was attributed to the effective secondary interaction between the boron atom and the incipient double bond in the norbonene resulting from the planar geometry of the catalyst. C-13 NMR chemical shifts of the coordinated dienophile carbonyl carbons with CBB compared to those of the non coordinated thiocinammates suggest a strong complexation with the catalyst.

Advanced lithium and lithium-ion rechargeable batteries for automotive applications

The worldwide demand for a clean and low-fuel-consuming transport promotes the development of safe, high energy and power electrochemical storage and conversion systems. Lithium-ion batteries (LIBs) are considered today the best technology for this application as demonstrated by the recent interest of automotive industry in hybrid (HEV) and electric vehicles (EV) based on LIBs. This thesis work, starting from the synthesis and characterization of electrode materials and the use of non-conventional electrolytes, demonstrates that LIBs with novel and safe electrolytes and electrode materials meet the targets of specific energy and power established by U.S.A. Department of Energy (DOE) for automotive application in HEV and EV. In chapter 2 is reported the origin of all chemicals used, the description of the instruments used for synthesis and chemical-physical characterizations, the electrodes preparation, the batteries configuration and the electrochemical characterization procedure of electrodes and batteries. Since the electrolyte is the main critical point of a battery, in particular in large- format modules, in chapter 3 we focused on the characterization of innovative and safe electrolytes based on ionic liquids (characterized by high boiling/decomposition points, thermal and electrochemical stability and appreciable conductivity) and mixtures of ionic liquid with conventional electrolyte. In chapter 4 is discussed the microwave accelerated sol–gel synthesis of the carbon- coated lithium iron phosphate (LiFePO 4 -C), an excellent cathode material for LIBs thanks to its intrinsic safety and tolerance to abusive conditions, which showed excellent electrochemical performance in terms of specific capacity and stability. In chapter 5 are presented the chemical-physical and electrochemical characterizations of graphite and titanium-based anode materials in different electrolytes. We also characterized a new anodic material, amorphous SnCo alloy, synthetized with a nanowire morphology that showed to strongly enhance the electrochemical stability of the material during galvanostatic full charge/discharge cycling. Finally, in chapter 6, are reported different types of batteries, assembled using the LiFePO 4 -C cathode material, different anode materials and electrolytes, characterized by deep galvanostatic charge/discharge cycles at different C-rates and by test procedures of the DOE protocol for evaluating pulse power capability and available energy. First, we tested a battery with the innovative cathode material LiFePO 4 -C and conventional graphite anode and carbonate-based electrolyte (EC DMC LiPF 6 1M) that demonstrated to surpass easily the target for power-assist HEV application. Given that the big concern of conventional lithium-ion batteries is the flammability of highly volatile organic carbonate- based electrolytes, we made safe batteries with electrolytes based on ionic liquid (IL). In order to use graphite anode in IL electrolyte we added to the IL 10% w/w of vinylene carbonate (VC) that produces a stable SEI (solid electrolyte interphase) and prevents the graphite exfoliation phenomenon. Then we assembled batteries with LiFePO 4 -C cathode, graphite anode and PYR 14 TFSI 0.4m LiTFSI with 10% w/w of VC that overcame the DOE targets for HEV application and were stable for over 275 cycles. We also assembled and characterized ―high safety‖ batteries with electrolytes based on pure IL, PYR 14 TFSI with 0.4m LiTFSI as lithium salt, and on mixture of this IL and standard electrolyte (PYR 14 TFSI 50% w/w and EC DMC LiPF 6 50% w/w), using titanium-based anodes (TiO 2 and Li 4 Ti 5 O 12 ) that are commonly considered safer than graphite in abusive conditions. The batteries bearing the pure ionic liquid did not satisfy the targets for HEV application, but the batteries with Li 4 Ti 5 O 12 anode and 50-50 mixture electrolyte were able to surpass the targets. We also assembled and characterized a lithium battery (with lithium metal anode) with a polymeric electrolyte based on poly-ethilenoxide (PEO 20 – LiCF 3 SO 3 +10%ZrO 2 ), which satisfied the targets for EV application and showed a very impressive cycling stability. In conclusion, we developed three lithium-ion batteries of different chemistries that demonstrated to be suitable for application in power-assist hybrid vehicles: graphite/EC DMC LiPF 6 /LiFePO 4 -C, graphite/PYR 14 TFSI 0.4m LiTFSI with 10% VC/LiFePO 4 -C and Li 4 T i5 O 12 /PYR 14 TFSI 50%-EC DMC LiPF 6 50%/LiFePO 4 -C. We also demonstrated that an all solid-state polymer lithium battery as Li/PEO 20 –LiCF 3 SO 3 +10%ZrO 2 /LiFePO 4 -C is suitable for application on electric vehicles. Furthermore we developed a promising anodic material alternative to the graphite, based on SnCo amorphous alloy.

Sviluppo dei materiali innovativi e studio della loro interazione con sistemi biologici e biomimetici

Nell’ambito della Chimica Sostenibile e dell’applicazione dei suoi principi per la salvaguardia dell’ambiente, il progetto di dottorato ha riguardato lo sviluppo di materiali innovativi e lo studio della loro interazione con sistemi biologici e biomimetici. In particolare l’attività si è focalizzata sulla sintesi di liquidi ionici ed indagini delle interazioni con membrane cellulari e sull’utilizzo ed isolamento di molecole da fonti rinnovabili. I liquidi ionici sono sali organici liquidi a temperature inferiori ai 100 °C; sono considerati promettenti solventi a ridotta tossicità, ma vanno chiarite a pieno le modalità di interazione con i sistemi biologici ed i meccanismi di tossicità. A questo scopo è stata impiegata una batteria di test bio-chimici, con saggi di fluorescenza e colorimetrici, che hanno permesso di discriminare le diverse tipologie di interazioni con varie strutture di membrana. Le informazioni raccolte sono servite per progettare sostanze meno dannose per le strutture cellulari, al fine di scegliere le funzionalità molecolari che consentano ai liquidi ionici di mantenere la loro attività ma di essere meno dannosi per l’ambiente. Per quanto riguarda l’utilizzo ed isolamento di molecole da fonte rinnovabili, si è utilizzata la tecnica della pirolisi per l’ottenimento di starting materials ed il loro impiego nella sintesi di chemicals in alternativa a composti derivanti da fonti fossili. La pirolisi tradizionale della cellulosa fornisce una molecola interessante, per semplicità denominata LAC, in quantità insufficienti ad un uso applicativo. Nell’ambito delle ricerche svolte è stato scoperto che la pirolisi condotta in presenza di catalizzatori meso-strutturati (MCM-41) drogati con metalli di transizione, fornisce buone quantità di LAC. LAC si è dimostrato promettente sia per la produzione di nuove molecole con possibili applicazioni nella chimica fine e farmaceutica, che come monomero per nuovi polimeri (copolimero ed omopolimero).

Tecniche elettroanalitiche-chemiometriche applicate alla tipizzazione dli oli monovarietali.

Il presente lavoro di tesi si propone di verificare se le tecniche elettrochimiche consentano di effettuare una classificazione del cultivar dell’olio. In particolare si vuole proporre un nuovo metodo per eseguire misure voltammetriche con un microelettrodo di platino direttamente in campioni di olio extravergine di oliva a cui sono aggiunti RTILs (Room Temperature Ionic Liquids) per rendere le matrici adeguatamente conducibili e quindi adatte per le analisi. I voltammogrammi ciclici dovranno poi essere elaborati attraverso diverse tecniche chemiometriche con l’obiettivo di discriminare i campioni sulla base della loro origine. Si studieranno quindi delle metodologie analitiche che potrebbero consentire di riconoscere l'origine delle olive usate per produrre un olio e di verificarne l'aderenza alle normative o ai disciplinari delle DOP e l'eventuale presenza di materiale estraneo. Le ricerche hanno l’obiettivo di individuare nell'olio il “fingerprint” da cui sarà possibile risalire alle varie cultivar di provenienza, identificando la composizione e l'origine dell'olio, ciò consentirà inoltre di difendere i consumatori da eventuali frodi commerciali.

Studio del metodo dell'assorbimento diretto della CO2 per l'analisi del radiocarbonio

L’attuale rilevanza rappresentata dalla stretta relazione tra cambiamenti climatici e influenza antropogenica ha da tempo posto l’attenzione sull’effetto serra e sul surriscaldamento planetario così come sull’aumento delle concentrazioni atmosferiche dei gas climaticamente attivi, in primo luogo la CO2. Il radiocarbonio è attualmente il tracciante ambientale per eccellenza in grado di fornire mediante un approccio “top-down” un valido strumento di controllo per discriminare e quantificare il diossido di carbonio presente in atmosfera di provenienza fossile o biogenica. Ecco allora che ai settori applicativi tradizionali del 14C, quali le datazioni archeometriche, si affiancano nuovi ambiti legati da un lato al settore energetico per quanto riguarda le problematiche associate alle emissioni di impianti, ai combustibili, allo stoccaggio geologico della CO2, dall’altro al mercato in forte crescita dei cosiddetti prodotti biobased costituiti da materie prime rinnovabili. Nell’ambito del presente lavoro di tesi è stato quindi esplorato il mondo del radiocarbonio sia dal punto di vista strettamente tecnico e metodologico che dal punto di vista applicativo relativamente ai molteplici e diversificati campi d’indagine. E’ stato realizzato e validato un impianto di analisi basato sul metodo radiometrico mediante assorbimento diretto della CO2 ed analisi in scintillazione liquida apportando miglioramenti tecnologici ed accorgimenti procedurali volti a migliorare le performance del metodo in termini di semplicità, sensibilità e riproducibilità. Il metodo, pur rappresentando generalmente un buon compromesso rispetto alle metodologie tradizionalmente usate per l’analisi del 14C, risulta allo stato attuale ancora inadeguato a quei settori applicativi laddove è richiesta una precisione molto puntuale, ma competitivo per l’analisi di campioni moderni ad elevata concentrazione di 14C. La sperimentazione condotta su alcuni liquidi ionici, seppur preliminare e non conclusiva, apre infine nuove linee di ricerca sulla possibilità di utilizzare questa nuova classe di composti come mezzi per la cattura della CO2 e l’analisi del 14C in LSC.

Kontinuierliche Kreuzkupplungsreaktionen in mikrostrukturierten Systemen

Im Rahmen dieser Arbeit wurden unterschiedliche Palladium-katalysierte Kreuzkupplungsreaktionen untersucht. Ein besonderes Augenmerk wurde dabei auf die Suzuki-Miyaura-Reaktion gelegt. Unter anderem aufgrund der langen Reaktionszeiten und der zweiphasigen Bedingungen ist diese Reaktionsklasse nur sehr schwer als kontinuierlicher Prozess zu etablieren. Vielen dieser Ansätze ist jedoch zu eigen, dass der große Vorteil der Mikroprozesstechnik, eine überlegene Kontrolle von Temperatur und Stofftransport, kaum ausgeschöpft wird. An diesem Punkt setzt diese Arbeit von technischer aus Seite an. Der zweite Schwerpunkt der Arbeit sind die prinzipiellen Untersuchungen an kontinuierlichen Flüssig-Flüssig-Zwei-Phasen-Reaktionen. Im Zuge des DBU-finanzierten Transkat-Projektes wurden hierbei anhand einfacher Veresterungsreaktionen grundlegende Kenntnisse zu Stofftransport, Grenzflächen und Phasentrennung innerhalb mikrostrukturierter Systeme gesammelt. Dank speziell angefertigter Glasmikroreaktoren von der Firma mikroglas chemtech GmbH war eine genaue optische und digitale Charakterisierung der Phasengrenzflächen möglich. Ein wichtiges Ergebnis war darüber hinaus, dass ionische Flüssigkeiten, als eigenständige Phasen verwendet, enorm zum Massentransfer und somit zur Reaktionsgeschwindigkeit beitragen können.

Screening, optimization and extraction of polyhydroxyalkanoates and peptidoglycan from Bacillus megaterium

Bioplastics are polymers (such as polyesters) produced from bacterial fermentations that are biodegradable and nonhazardous. They are produced by a wide variety of bacteria and are made only when stress conditions allow, such as when nutrient levels are low, more specifically levels of nitrogen and oxygen. These stress conditions cause certain bacteria to build up excess carbon deposits as energy reserves in the form of polyhydroxyalkanoates (PHAs). PHAs can be extracted and formed into actual plastic with the same strength of conventional, synthetic-based plastics without the need to rely on foreign petroleum. The overall goal of this project was to select for a bacteria that could grow on sugars found in the lignocellulosic biomass, and get the bacteria to produce PHAs and peptidoglycan. Once this was accomplished the goal was to extract PHAs and peptidoglycan in order to make a stronger more rigid plastic, by combing them into a co-polymer. The individual goals of this project were to: (1) Select and screen bacteria that are capable of producing PHAs by utilizing the carbon/energy sources found in lignocellulosic biomass; (2) Maximize the utilization of those sugars present in woody biomass in order to produce optimal levels of PHAs. (3) Use room temperature ionic liquids (RTILs) in order to separate the cell membrane and peptidoglycan, allowing for better extraction of PHAs and more intact peptidoglycan. B. megaterium a Gram-positive PHA-producing bacterium was selected for study in this project. It was grown on a variety of different substrates in order to maximize both its growth and production of PHAs. The optimal conditions were found to be 30°C, pH 6.0 and sugar concentration of either 30g/L glucose or xylose. After optimal growth was obtained, both RTILs and enzymatic treatments were used to break the cell wall, in order to extract the PHAs, and peptidoglycan. PHAs and peptidoglycan were successfully extracted from the cell, and will be used in the future to create a new stronger co-polymer. Peptidoglycan recovery yield was 16% of the cells’ dry weight.

The old problems of glass and the glass transition, and the many new twists.

In this paper I review the ways in which the glassy state is obtained both in nature and in materials science and highlight a "new twist"--the recent recognition of polymorphism within the glassy state. The formation of glass by continuous cooling (viscous slowdown) is then examined, the strong/fragile liquids classification is reviewed, and a new twist-the possibility that the slowdown is a result of an avoided critical point-is noted. The three canonical characteristics of relaxing liquids are correlated through the fragility. As a further new twist, the conversion of strong liquids to fragile liquids by pressure-induced coordination number increases is demonstrated. It is then shown that, for comparable systems, it is possible to have the same conversion accomplished via a first-order transition within the liquid state during quenching. This occurs in the systems in which "polyamorphism" (polymorphism in the glassy state) is observed, and the whole phenomenology is accounted for by Poole's bond-modified van der Waals model. The sudden loss of some liquid degrees of freedom through such weak first-order transitions is then related to the polyamorphic transition between native and denatured hydrated proteins, since the latter are also glass-forming systems--water-plasticized, hydrogen bond-cross-linked chain polymers (and single molecule glass formers). The circle is closed with a final new twist by noting that a short time scale phenomenon much studied by protein physicists-namely, the onset of a sharp change in d/dT ( is the Debye-Waller factor)--is general for glass-forming liquids, including computer-simulated strong and fragile ionic liquids, and is closely correlated with the experimental glass transition temperature. The latter thus originates in strong anharmonicity in certain components of the vibrational density of states, which permits the system to access the multiple minima of its configuration space. The connection between the anharmonicity in these modes, vibrational localization, the Kauzmann temperature, and the fragility of the liquid is proposed as the key problem in glass science.