Production of lipase by extractive fermentation with ionic liquids

Novos processos fermentativos, designados por processos de Fermentação Extractiva, são caracterizados por apresentarem etapas de produção e extracção em simultâneo. A extracção líquido-líquido como técnica de separação é amplamente usado na indústria química pela sua simplicidade, baixo custo e facilidade de extrapolação de escala. No entanto o uso de solventes orgânicos nestes processos potencia os riscos ocupacionais e ambientais. Neste contexto, o uso de sistemas de duas fases aquosas baseados em líquidos iónicos, apresenta-se como uma técnica eficaz para a separação e purificação de produtos biológicos. Este trabalho apresenta um estudo integrado sobre o uso de líquidos iónicos não aromáticos foram determinados. A capacidade para a formação de sistemas de duas fases foi estudada para uma vasta gama de líquidos iónicos hidrofílicos com diferentes aniões, catiões e cadeias alqúilicas. A capacidade de separação e purificação de um largo conjunto de líquidos iónicos foi posteriormente investigada, recorrendo-se ao uso de várias biomoléculas modelo de diferentes graus de complexidade, um amino-acido (L-triptofano) e duas enzimas lipolíticas (enzima produzida pela bactéria Bacillus sp. e Candida antarctica lipase B – CaLB). Esta última foi ainda usada para um estudo de biocompatibilidade, tendo sido determinado o efeito de diferentes LIs hidrofílicos na sua actividade enzimática. Este trabalho mostra um estudo ecotoxicológico duma vasta gama de líquidos iónicos e espécies aquáticas, inseridas em diversos níveis tróficos. A bioacumulação foi investigada através do estudo dos coeficientes de distribuição 1-octanol-água (Dow).

Economical and operational optimization of CALB immobilization on MP1000 for amidation of olein fatty acids with ethanolamine

Dissertação de Mestrado , Engenharia Biológica, Faculdade de Engenharia de Recursos Naturais, Universidade do Algarve, 2008

Evolutionary Origin and Maintenance of Sociality in the Small Carpenter Bees

Many arthropods exhibit behaviours precursory to social life, including adult longevity, parental care, nest loyalty and mutual tolerance, yet there are few examples of social behaviour in this phylum. The small carpenter bees, genus Ceratina, provide important insights into the early stages of sociality. I described the biology and social behaviour of five facultatively social species which exhibit all of the preadaptations for successful group living, yet present ecological and behavioural characteristics that seemingly disfavour frequent colony formation. These species are socially polymorphic with both / solitary and social nests collected in sympatry. Social colonies consist of two adult females, one contributing both foraging and reproductive effort and the second which remains at the nest as a passive guard. Cooperative nesting provides no overt reproductive benefits over solitary nesting, although brood survival tends to be greater in social colonies. Three main theories explain cooperation among conspecifics: mutual benefit, kin selection and manipulation. Lifetime reproductive success calculations revealed that mutual benefit does not explain social behaviour in this group as social colonies have lower per capita life time reproductive success than solitary nests. Genetic pedigrees constructed from allozyme data indicate that kin selection might contribute to the maintenance of social nesting -, as social colonies consist of full sisters and thus some indirect fitness benefits are inherently bestowed on subordinate females as a result of remaining to help their dominant sister. These data suggest that the origin of sociality in ceratinines has principal costs and the great ecological success of highly eusociallineages occurred well after social origins. Ecological constraints such as resource limitation, unfavourable weather conditions and parasite pressure have long been considered some of the most important selective pressures for the evolution of sociality. I assessed the fitness consequences of these three ecological factors for reproductive success of solitary and social colonies and found that nest sites were not limiting, and the frequency of social nesting was consistent across brood rearing seasons. Local weather varied between seasons but was not correlated with reproductive success. Severe parasitism resulted in low reproductive success and total nest failure in solitary nests. Social colonies had higher reproductive success and were never extirpated by parasites. I suggest that social nesting represents a form of bet-hedging. The high frequency of solitary nests suggests that this is the optimal strategy when parasite pressure is low. However, social colonies have a selective advantage over solitary nesting females during periods of extreme parasite pressure. Finally, the small carpenter bees are recorded from all continents except Antarctica. I constructed the first molecular phylogeny of ceratinine bees based on four gene regions of selected species covering representatives from all continents and ecological regions. Maximum parsimony and Bayesian Inference tree topology and fossil dating support an African origin followed by an Old World invasion and New World radiation. All known Old World ceratinines form social colonies while New World species are largely solitary; thus geography and phylogenetic inertia are likely predictors of social evolution in this genus. This integrative approach not only describes the behaviour of several previously unknown or little-known Ceratina species, bu~ highlights the fact that this is an important, though previously unrecognized, model for studying evolutionary transitions from solitary to social behaviour.

ORGANOSILICON BIOTECHNOLOGY: A BIO-INSPIRED APPROACH TO THE HYDROLYSIS OF ALKOXYSILANES and THE LIPASE-CATALYZED SYNTHESIS OF SILOXANE-CONTAINING POLYESTERS AND POLYAMIDES

The first part of this thesis studied the capacity of amino acids and enzymes to catalyze the hydrolysis and condensation of tetraethoxysilane and phenyltrimethoxysilane. Selected amino acids were shown to accelerate the hydrolysis and condensation of tetraethoxysilane under ambient temperature, pressure and at neutral pH (pH 7±0.02). The nature of the side chain of the amino acid was important in promoting hydrolysis and condensation. Several proteases were shown to have a capacity to hydrolyze tri- and tet-ra- alkoxysilanes under the same mild reaction conditions. The second part of this thesis employed an immobilized Candida antarctica lipase B (Novozym-435, N435) to produce siloxane-containing polyesters, polyamides, and polyester amides under solvent-free conditions. Enzymatic activity was shown to be temperature dependent, increasing until enzyme denaturation became the dominant pro-cess, which typically occurred between 120-130ᵒC. The residual activity of N435 was, on average, greater than 90%, when used in the synthesis of disiloxane-containing polyesters, regardless of the polymerization temperature except at the very highest temperatures, 140-150ᵒC. A study of the thermal tolerance of N435 determined that, over ten reaction cycles, there was a decrease in the initial rate of polymerization with each consecutive use of the catalyst. No change in the degree of monomer conversion after a 24 hour reaction cycle was found.

Lipase-Mediated Enzymatic Catalysis For The Synthesis of New Chiral Polymers

Immobilized lipase B from Candida antarctica (N435) was investigated as a potential biocatalyst to generate silicone-based chiral polymers from monomers derived from the enzymatic dihydroxylation of bromobenzene. Several conditions and parameters have been investigated for this purpose and lipase transesterification preference to each of the free secondary alcohols in the chiral monomers was documented. The N435 was challenged with a series of substrates where the free alcohol moieties were systematically protected in order to study the substrate preference(s) for the transesterification reactions.

Cyclotetrasiloxane Frameworks for the Chemoenzymatic Synthesis of Oligoesters

Immobilized lipase B from Candida antarctica (Novozym® 435, N435) was utilized as part of a chemoenzymatic strategy for the synthesis of branched polyesters based on a cyclotetrasiloxane core in the absence of solvent. Nuclear magnetic resonance spectroscopy and matrix-assisted laser desorption ionization time-of-flight mass spectrometry were utilized to monitor the reactions between tetraester cyclotetrasiloxanes and aliphatic diols. The enzyme-mediated esterification reactions can achieve 65– 80% consumption of starting materials in 24–48 h. Longer reaction times, 72–96 h, resulted in the formation of cross-linked gel-like networks. Gel permeation chromatography of the polymers indicated that the masses were Mw ¼ 11 400, 13 100, and 19 400 g mol 1 for the substrate pairs of C7D4 ester/ octane-1,8-diol, C10D4 ester/pentane-1,5-diol and C10D4 ester/octane-1,8-diol respectively, after 48 h. Extending the polymerization for an additional 24 h with the C10D4 ester/octane-1,8-diol pair gave Mw ¼ 86 800 g mol 1. To the best of our knowledge this represents the first report using lipase catalysis to produce branched polymers that are built from a cyclotetrasiloxane core.

Le modèle antarctique

Il existe actuellement des différends frontaliers entre les États circumpolaires-Nord qui entravent la bonne gouvernance de la région. Afin d'identifier des pistes de solution pour améliorer la gouvernance en Arctique et faciliter la coopération, nous analysons le processus qui a mené, à l'autre pôle, à la création du Traité sur l’Antarctique de 1959. Nous utilisons une approche néolibérale institutionnelle comme cadre théorique ainsi qu’une méthodologie basée sur le traçage de processus. Nous avons identifié quatre variables (les intérêts des États, le contexte politique, le leadership, et l’Année géophysique internationale [AGI]) que nous avons étudiés au cours de trois périodes historiques et dont l’évolution a contribué à la formation du régime antarctique. Les étapes menant à l'élaboration d'un régime identifié dans la théorie institutionnaliste dynamique de Frischmann s'appliquent aux différentes périodes que nous avons déterminées. La première période historique correspond à l'identification du problème d'interdépendance. Les États ont d’abord tenté d’assouvir leurs intérêts particuliers envers le continent antarctique par l’intermédiaire de stratégies individuelles. Le statut imprécis du continent antarctique caractérisé par des revendications territoriales conflictuelles, la volonté des superpuissances d’effectuer d’éventuelles revendications territoriales et le leadership exercé par les États-Unis ont toutefois permis de faire évoluer les États vers des stratégies coopératives. La deuxième période historique correspond à l'étape de la négociation de l’entente. Le leadership d’un groupe de scientifiques et des États-Unis ainsi que le contexte politique de la Guerre froide ont favorisé la cohésion des États intéressés par les affaires antarctiques et leur investissement dans une coopération multilatérale souple dans le cadre d’un événement ponctuel, l’AGI. La troisième période correspond à la mise en oeuvre de l’entente, soit l’AGI. Lors de cet événement, une entente tacite, un engagement d’honneur, qui suspendait les considérations territoriales en Antarctique, a été respectée par les parties. La coopération dans le cadre de l’AGI s’est avérée un succès. Les États se sont montrés satisfaits des gains que comportait la coopération scientifique dans un environnement pacifique. L’AGI a permis aux États d’expérimenter la coopération multilatérale et d’observer le comportement des autres participants au sein de l’institution. En raison de la satisfaction des États vis-à-vis la coopération dans le cadre de l'entente souple, l'élaboration du traité de 1959 a été facilitée. Notre étude nous a permis d’identifier des éléments clefs qui ont favorisé la formation du régime antarctique et qui pourraient servir d’inspiration pour l'élaboration de futurs régimes. La présence d’un leadership tout au long du processus a permis de promouvoir les avantages de la coopération sous différentes formes et de renforcer les relations entre les États présents dans les affaires antarctiques. La réalisation d'une entente souple dans le cadre de l'AGI, centrée autour des intérêts communs aux parties, comprenant un engagement d’honneur, permettant de mettre de côté les points de discorde, ont aussi été des points centraux dans la réussite et la poursuite de la coopération au-delà de l’AGI. Cette approche graduelle allant du droit souple vers le droit contraignant est sans aucun doute l’élément le plus important ayant facilité le rapprochement des différentes parties et éventuellement la création du Traité sur l’Antarctique.

Geochemistry of core sediment from antarctic region

Southern Ocean (SO) is the fourth largest Ocean comprising the southern portions of the Atlantic Ocean, Indian Ocean and Pacific Ocean. Sediment core sample (660 34’S and 580 40’E)was collected onboard O.R.V Sagar Nidhi from January to March 2010 in the Fourth Southern Ocean expedition cruise launched by the National Centre for Antarctic and Ocean Research, Goa . Sedimentary records from this area reveal the sensitivity and climatic variability’s of the region over a large time scale. Organic matter (OM) and textural behaviour of the samples were analyzed and processed concurrently. Distribution of OM, Total Organic Carbon (TOC), Protein, Lipid and Carbohydrate along with the trace metal was highlighted. Textural variation was in the array of Sand >Clay >Silt. Sand content ranges from 30.29% to 80.11%. The order of relative distribution of OM was Lipid >Protein > TOC > Carbohydrate. The average concentrations of TOC, Protein, Lipid and Carbohydrate were 2.2 mg/g, 1.2 mg/g, 3.3 mg/g and 1.1mg/g respectively. Protein to carbohydrate ratio and lipid to carbohydrate ratio were also encountered to understand the respective freshness and nutritional quality of the sediments. Trace metal distribution showed the average concentration was maximum for Mn and minimum for Co.

¿Antártida Argentina?: incidencia del discurso formal del Instituto Antártico Argentino en la implementación del discurso práctico peronista durante el periodo comprendido entre 1945 y 1960

Entre 1945 y 1960 la Argentina de Juan Domingo Perón, condicionada para el nacionalismo, participa de la internacionalización de la Antártida y con ello, inicia una contienda simbólica con Gran Bretaña por ‘representar’ un espacio que considera parte de su ‘Nueva Argentina’. Evocando el ‘razonamiento geográfico’ y el patriotismo más antiguo de los argentinos, Perón instrumentaliza las bases científicas y cartografías australes, para implementar su discurso. Irónicamente, el control interno y discursivo que le permitiría su vínculo con la Armada, también sería la causa de su caída.

Las pretensiones geoestratégicas de Colombia luego de su adhesión al Tratado Antártico en 1989

El interés de esta investigación es analizar los antecedentes y desarrollo del interés colombiano en la Antártida, así como la relación de cooperación colombo-chilena y la importancia geoestratégica del continente blanco, a partir de diferentes conceptos: geoestrategia, política exterior colombiana y cooperación internacional. Teniendo en cuenta que la Antártida refleja los intereses geoestratégicos de muchos países desde comienzos del siglo XX, Colombia no puede dejar de lado sus aspiraciones en el Sistema Internacional y por esta razón decide ingresar al Tratado Antártico en 1989, sin embargo, es necesario definir cuáles son las pretensiones geoestratégicas del país suramericano en este territorio y precisar la probabilidad de su cambio de estatus en el mismo que le permita obtener provechos materiales de forma unilateral en materia de recursos, actividad diplomática, política exterior y posición geográfica.

A new perspective on Southern hemisphere storm-tracks.

A detailed view of Southern Hemisphere storm tracks is obtained based on the application of filtered variance and modern feature-tracking techniques to a wide range of 45-yr European Centre for Medium-Range Weather Forecasts (ECMWF) Re-Analysis (ERA-40) data. It has been checked that the conclusions drawn in this study are valid even if data from only the satellite era are used. The emphasis of the paper is on the winter season, but results for the four seasons are also discussed. Both upper- and lower-tropospheric fields are used. The tracking analysis focuses on systems that last longer than 2 days and are mobile (move more than 1000 km). Many of the results support previous ideas about the storm tracks, but some new insights are also obtained. In the summer there is a rather circular, strong, deep high-latitude storm track. In winter the high-latitude storm track is more asymmetric with a spiral from the Atlantic and Indian Oceans in toward Antarctica and a subtropical jet–related lower-latitude storm track over the Pacific, again tending to spiral poleward. At all times of the year, maximum storm activity in the higher-latitude storm track is in the Atlantic and Indian Ocean regions. In the winter upper troposphere, the relative importance of, and interplay between, the subtropical and subpolar storm tracks is discussed. The genesis, lysis, and growth rate of lower-tropospheric winter cyclones together lead to a vivid picture of their behavior that is summarized as a set of overlapping plates, each composed of cyclone life cycles. Systems in each plate appear to feed the genesis in the next plate through downstream development in the upper-troposphere spiral storm track. In the lee of the Andes in South America, there is cyclogenesis associated with the subtropical jet and also, poleward of this, cyclogenesis largely associated with system decay on the upslope and regeneration on the downslope. The genesis and lysis of cyclones and anticyclones have a definite spatial relationship with each other and with the Andes. At 500 hPa, their relative longitudinal positions are consistent with vortex-stretching ideas for simple flow over a large-scale mountain. Cyclonic systems near Antarctica have generally spiraled in from lower latitudes. However, cyclogenesis associated with mobile cyclones occurs around the Antarctic coast with an interesting genesis maximum over the sea ice near 150°E. The South Pacific storm track emerges clearly from the tracking as a coherent deep feature spiraling from Australia to southern South America. A feature of the summer season is the genesis of eastward-moving cyclonic systems near the tropic of Capricorn off Brazil, in the central Pacific and, to a lesser extent, off Madagascar, followed by movement along the southwest flanks of the subtropical anticyclones and contribution to the “convergence zone” cloud bands seen in these regions.

An overview of Earth's global electric circuit and atmospheric conductivity

The Earth’s global atmospheric electric circuit depends on the upper and lower atmospheric boundaries formed by the ionosphere and the planetary surface. Thunderstorms and electrified rain clouds drive a DC current (∼1 kA) around the circuit, with the current carried by molecular cluster ions; lightning phenomena drive the AC global circuit. The Earth’s near-surface conductivity ranges from 10−7 S m−1 (for poorly conducting rocks) to 10−2 S m−1 (for clay or wet limestone), with a mean value of 3.2 S m−1 for the ocean. Air conductivity inside a thundercloud, and in fair weather regions, depends on location (especially geomagnetic latitude), aerosol pollution and height, and varies from ∼10−14 S m−1 just above the surface to 10−7 S m−1 in the ionosphere at ∼80 km altitude. Ionospheric conductivity is a tensor quantity due to the geomagnetic field, and is determined by parameters such as electron density and electron–neutral particle collision frequency. In the current source regions, point discharge (coronal) currents play an important role below electrified clouds; the solar wind-magnetosphere dynamo and the unipolar dynamo due to the terrestrial rotating dipole moment also apply atmospheric potential differences. Detailed measurements made near the Earth’s surface show that Ohm’s law relates the vertical electric field and current density to air conductivity. Stratospheric balloon measurements launched from Antarctica confirm that the downward current density is ∼1 pA m−2 under fair weather conditions. Fortuitously, a Solar Energetic Particle (SEP) event arrived at Earth during one such balloon flight, changing the observed atmospheric conductivity and electric fields markedly. Recent modelling considers lightning discharge effects on the ionosphere’s electric potential (∼+250 kV with respect to the Earth’s surface) and hence on the fair weather potential gradient (typically ∼130 V m−1 close to the Earth’s surface. We conclude that cloud-to-ground (CG) lightning discharges make only a small contribution to the ionospheric potential, and that sprites (namely, upward lightning above energetic thunderstorms) only affect the global circuit in a miniscule way. We also investigate the effects of mesoscale convective systems on the global circuit.

Improvements to the accuracy of measurements of NO2 by zenith-sky visible spectrometers II: errors in zero using a more complete chemical model

Using a flexible chemical box model with full heterogeneous chemistry, intercepts of chemically modified Langley plots have been computed for the 5 years of zenith-sky NO2 data from Faraday in Antarctica (65°S). By using these intercepts as the effective amount in the reference spectrum, drifts in zero of total vertical NO2 were much reduced. The error in zero of total NO2 is ±0.03×1015 moleccm−2 from one year to another. This error is small enough to determine trends in midsummer and any variability in denoxification between midwinters. The technique also suggests a more sensitive method for determining N2O5 from zenith-sky NO2 data.

Geomorphological Map of Ribbed Moraines on the Dubawnt Lake Palaeo-Ice Stream Bed: A Signature of Ice Stream Shut-down?

The beds of active ice streams in Greenland and Antarctica are largely inaccessible, hindering a full understanding of the processes that initiate, sustain and inhibit fast ice flow in ice sheets. Detailed mapping of the glacial geomorphology of palaeo-ice stream tracks is, therefore, a valuable tool for exploring the basal processes that control their behaviour. In this paper we present a map that shows detailed glacial geomorphology from a part of the Dubawnt Lake Palaeo-Ice Stream bed on the north-western Canadian Shield (Northwest Territories), which operated at the end of the last glacial cycle. The map (centred on 63 degrees 55 '' 42'N, 102 degrees 29 '' 11'W, approximate scale 1:90,000) was compiled from digital Landsat Enhanced Thematic Mapper Plus satellite imagery and digital and hard-copy stereo-aerial photographs. The ice stream bed is dominated by parallel mega-scale glacial lineations (MGSL), whose lengths exceed several kilometres but the map also reveals that they have, in places, been superimposed with transverse ridges known as ribbed moraines. The ribbed moraines lie on top of the MSGL and appear to have segmented the individual lineaments. This indicates that formation of the ribbed moraines post-date the formation of the MSGL. The presence of ribbed moraine in the onset zone of another palaeo-ice stream has been linked to oscillations between cold and warm-based ice and/or a patchwork of cold-based areas which led to acceleration and deceleration of ice velocity. Our hypothesis is that the ribbed moraines on the Dubawnt Lake Ice Stream bed are a manifestation of the process that led to ice stream shut-down and may be associated with the process of basal freeze-on. The precise formation of ribbed moraines, however, remains open to debate and field observation of their structure will provide valuable data for formal testing of models of their formation.