Sistemas aquosos bifásicos uma ferramenta sustentável para a extração de ácido clavulânico a partir de diferentes fontes

O ácido clavulânico (AC) é um inibidor de β-lactamases que tem vindo a ser largamente utilizado na área médica. Embora seja de extrema importância, o desenvolvimento de processos alternativos de produção e purificação é ainda insignificante, sendo fundamental o estudo de técnicas de extração mais biocompatíveis, como os Sistemas Aquosos Bifásicos (SABs). Assim, este trabalho objetivou o estudo de Sistemas Aquosos Bifásicos baseados em polímeros como uma ferramenta alternativa para a extração de AC. Foram testados dois SPAB compostos por Polietileno Glicol (PEG) com massa molecular (M) de 4000 g/mol e Poliacrilato de Sódio de 8000 g/mol, nos quais foi alterado o eletrólito indutor da formação de fases, em particular, sulfato de sódio (Na2SO4,) e cloreto de sódio (NaCl). Ademais, este trabalho visou também avaliar a eficiência de extração do AC, bem como compreender o efeito dos contaminantes no processo de migração. Para tal, foi avaliada a extração do AC a partir de três fontes distintas: solução pura (99,9%); solução comercial (60%); diretamente a partir do sobrenadante de um meio fermentando de Streptomyces clavuligerus. Os resultados obtidos demonstraram que independentemente da fonte inicial do AC, ambos os SABs poliméricos promoveram uma partição preferencial do AC para a fase rica em PEG, sendo o coeficiente de partição maior nos sistemas com Na2SO4 do que com NaCl. Após identificar a grande capacidade de partição de AC, o SAB com PEG/NaPA/Na2SO4 foi também utilizado para avaliar a partição de proteínas presente no meio fermentado, sendo também obtida uma preferencial partição destas para a fase rica em PEG. Assim, apesar da baixa capacidade de purificação de AC frente a proteínas contaminantes, os SABs estudados demonstraram que podem ser uma técnica alternativa sustentável e bastante econômica para uma etapa inicial de clarificação/concentração de bioprodutos a partir de caldos fermentados

Temperature and solvent effects on photoluminescence of tryptophan solutions

The fluorescence of tryptophan solutions in glycerol, water and ethanol was measured as a function of temperature using steady state and time-resolved techniques. The non-radiative rate constants responsible for the variation of fluorescence quantum yields with temperature are evaluated. Spectral shifts occurring on the nanosecond scale are interpreted in terms of an inversion of 1 L b and 1 L a states produced by solvent perturbation.info:eu-repo/semantics/publishedVersio

A stable liquid–liquid extraction system for clavulanic acid using polymer-based aqueous two-phase systems

The partitioning of Clavulanic Acid (CA) in a novel inexpensive and stable aqueous two-phase system (ATPS) composed by poly(ethylene glycol) (PEG) and sodium polyacrylate (NaPA) has been studied. The aqueous two-phase systems are formed by mixing both polymers with a salt (NaCl or Na2SO4) and an aqueous solution of CA. The stability of CA on the presence of both polymers was investigated and it was observed that these polymers do not degrade the biomolecule. The effect of PEG-molecular size, polymer concentrations on the commercial CA partitioning has been studied, at 25 C. The data showed that commercial CA was preferentially partitioned for the PEG-rich phase with a partition coefficient (KCA) between 1 and 12 in the PEG/NaPA aqueous two phase systems supplemented with NaCl and Na2SO4. The partition to the PEG phase was increased in the systems with high polymer concentrations. Furthermore, Na2SO4 caused higher CA preference for the PEG-phase than NaCl. The systems having a composition with 10 wt.% of PEG4000, 20 wt.% of NaPA8000 and 6 wt.% of Na2SO4 were selected as the optimal ones in terms of recovery of CA from fermented broth of Streptomyces clavuligerus. The partitioning results (KCA = 9.15 ± 1.06) are competitive with commercial extraction methods of CA (KCA = 11.91 ± 2.08) which emphasizes that the system PEG/NaPA/Na2SO4 can be used as a new process to CA purification/concentrationFinancial support is gratefully acknowledged from FAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo, Sao Paulo, Brazil) and from CNPq (National Council for Scientific and Technological Development, Brasilia, Brazil) and CAPES (Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior)

Tuning the ionic character of sodium dodecyl sulphate via counter-ion binding: an experimental and computational study

Solutions of surfactants exhibit remarkable features, such as a tunable amphiphilic character, which can further be varied for ionic surfactants through variations in their Coulombic interactions. These properties are very useful in many industrial applications such as in extraction, purification, and formulation processes, as detergents, wetting agents, or emulsifiers. Rather unexpectedly, the addition of tetrabutylammonium chloride ([N4,4,4,4]Cl) to solutions of the ionic surfactant of sodium dodecyl sulphate (SDS) results in the appearance of a phase transition above the lower critical solution temperature (LCST), a property usually associated with non-ionic surfactants. The aim of this study is to provide a detailed nanoscopic scenario on the interaction between SDS micelles and [N4,4,4,4]Cl moieties to better understand the nature of the LCST cloud point and how to confer it to a given ionic surfactant system. A coarse-grained molecular dynamics (CG-MD) computational framework, under the latest MARTINI 3.0 force field, was developed and validated using available literature data. The impact of [N4,4,4,4]Cl concentration in the phase of SDS micellar aqueous solutions was then characterized and compared using experimental results. Specifically, dynamic light scattering (DLS) measurements and small-angle X-ray scattering (SAXS) profiles were obtained at different [N4,4,4,4]+/[DS]- molar ratios (from 0.0 to 1.0) and compared with the CG-MD results. A good agreement between computer simulations and experimental findings was obtained, reinforcing the suitability of GC-MD to simulate complex phase behaviors. When the [N4,4,4,4]+/[DS]- molar ratio is 0.5, the system yielded clusters of enclosed small [DS]- aggregates. Thus, the CG-MD simulations showed the formation of mixed [DS]- and [N4,4,4,4]+ aggregates with [N4,4,4,4]+ cations acting as a bridge between small [DS]- micelles. The CG-MD simulation framework developed in this work captured the role of [N4,4,4,4]+ in the micellar phase transition whilst improving the results obtained with preceding computer models for which the limitations on capturing SDS and [N4,4,4,4]Cl mixtures in aqueous solutions are also shown in detail.publishe

Enhanced oil recovery under laboratory conditions using biosurfactant-producing microorganisms

Oil recovery comprises a primary phase, which produces oil using the natural pressure drive of the reservoir, and a secondary phase, which includes the injection of water to improve the flow of oil to the wellhead [1,2]. While primary recovery produces 5-10% of the original oil in place, recovery efficiencies in the secondary stage vary from 10% to 40% [1]. Most of the unrecovered oil (up to two-thirds of the total oil reserves) is trapped in the reservoir pores by high capillary forces [2]. Microbial Enhanced Oil Recovery (MEOR) is a tertiary oil recovery process where microorganisms and their metabolites are used to retrieve unrecoverable oil from mature reservoirs. Stimulation of biosurfactant production by indigenous or injected microorganisms can reduce the capillary forces that retain the oil into the reservoir. In this work, a sand pack column model was designed to simulate the oil recovery operations in oil reservoirs and evaluate the mobilization of residual oil by microorganisms. Three Bacillus subtilis strains (309, 311 and 573), previously isolated from crude oil samples, were used in this study. They grow and produce extracellular biosurfactants at 40ºC under anaerobic conditions in medium supplemented with hydrocarbons. Biosurfactants produced by those isolates reduce the surface tension of water from 72 to 30 mN/m, exhibit emulsifying activity and are not affected by exposure to high temperatures (121ºC) which makes them good candidates for application in biosurfactant mediated MEOR. Acrylic columns (250 ml) packed with acid washed sand were first flooded with water, after that saturated with paraffin, and then washed with water to remove the excess of paraffin. Afterwards, the isolates were injected into the columns with the optimized medium and incubated at 40ºC. After 14 days, the columns were flooded with water and the additional oil recovery (AOR) was calculated as the percentage of paraffin recovered. AOR using B. subtilis 309, 311 and 573 was 35.0 ± 1.0 %, 23.5 ± 1.2 % and 19.8 ± 1.9 %, respectively. The results obtained suggest that stimulation of biosurfactant production by these strains in the oil reservoir can contribute to mobilize entrapped oi

Isolation and study of microorganisms from oil samples for application in Microbial Enhanced Oil Recovery

"Author's personal copy"Microbial Enhanced Oil Recovery (MEOR) is potentially useful to increment oil recovery from a reservoir beyond primary and secondary recovery operations using microorganisms and their metabolites. Stimulation of bacterial growth for biosurfactant production and degradation of heavy oil fractions by indigenous microorganisms can enhance the fluidity and reduce the capillary forces that retain the oil into the reservoir. MEOR offers major advantages over conventional EOR, namely low energy consumption and independence of the price of crude oil. In this work, the isolation and identification of microorganisms capable of producing biosurfactants and promote degradation of long-chain n-alkanes under conditions existent in oil reservoirs were addressed. Among the isolated microorganisms, five Bacillus strains were able to produce extracellular biosurfactants at 40 C under anaerobic conditions in medium supplemented with hydrocarbons. Three isolates were selected as the higher biosurfactant producers. The obtained biosurfactants reduced the surface tension of water from 72 to 30 mN/m, exhibited emulsifying activity and were not affected by exposure to high temperatures (121 C). These characteristics make them good candidates for use at conditions usually existing in oil reservoirs. Furthermore, it was here shown for the first time that Bacillus strains were able to degrade large alkyl chains and reduce the viscosity of hydrocarbon mixtures under anaerobic conditions. The results obtained show that the isolated microorganisms are promising candidates for the development of enhanced oil recovery processes.This work was supported by PARTEX OIL AND GAS. Jorge F. B. Pereira acknowledges the financial support from Fundacao para a Ciencia e a Tecnologia through doctoral research grant SFRH/BD/60228/2009

Using coarse-grained molecular dynamics to understand the effect of ionic liquids on the aggregation of Pluronic copolymer solutions

This study is aimed to enhance the understanding of the interaction between ionic liquids (ILs) and non-ionic Pluronic triblock copolymers in aqueous two-phase micellar systems (ATPMS) used for the selective separation/purification of hydrophobic biomolecules. The ILs allow a precise control of the cloud point phase separation temperature (CPT), particularly important when the stability of the molecule is highly dependent on temperature. The effect of choline-based ILs, with two different counter-anions, chloride and hexanoate, was evaluated using molecular dynamics simulations (MD) for F-68 and L-35 Pluronic aqueous solutions. The simulations revealed the role played by the anions during the Pluronic self-assembly, with choline chloride hindering Pluronic aggregation and the choline hexanoate favouring micelle formation and coalescence, in agreement with the experimental data. A detailed study of the accessible surface area of Pluronic showed a progressive dehydration of the Pluronic hydrophilic micelle corona in choline hexanoate mixtures promoting inter-micelle interactions and, consequently, micelle coalescence. With the addition of choline hexanoate, it was observed that the hydrophilic segments, which form the micelle corona, twisted towards the Pluronic micelle core. The electrostatic interaction is also shown to play a key role in this IL–Pluronic aqueous solution, as the hexanoate anions are accommodated in the Pluronic micelle core, while the choline cations are hosted by the Pluronic micelle corona, with the ions interacting with each other during the self-assembly process. In addition, a comparison study of F-68 and L-35 aqueous solutions shows that the IL impact depends on the length of the Pluronic hydrophilic segment. This work provides a realistic microscopic scenario of the complex interactions between Pluronic copolymers and ILs.This work was developed within the scope of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020 & UIDP/50011/2020, financed by national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement. The authors acknowledge the research contract under the project CENTRO-01-0145- FEDER-000005: SusPhotoSolutions: Soluções Fotovoltaicas Sustentáveis. G. Pérez-Sánchez and N. Schaeffer acknowledge the national funds (OE), through FCT – Fundação para a Ciência e a Tecnologia, I. P., in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29th, changed by Law 57/2017, of July 19th. A. M. Lopes acknowledges the support from the State of São Paulo Research Foundation (FAPESP/Brazil, processes #2017/10789-1 and #2018/10799-0). J. F. B. Pereira also acknowledges FAPESP through the project 2014/16424-7.publishe

Microbial degradation of hydrocarbons and its applications to enhanced oil recovery at lab scale

The renewed interest in Enhanced Oil Recovery (EOR) techniques as a consequence of the current oil prices is boosting the development of the Microbial Enhanced Oil Recovery (MEOR). This technique is useful to recover incremental oil from a reservoir beyond primary and secondary recovery operations and can be carried by the injection of exogenous or stimulation of indigenous microorganisms. This last approach is here investigated. In this work we address the isolation and identification of microorganisms from Brazilian heavy oil samples capable of producing biosurfactants and to promote the degradation of heavy oil fractions, in particular long-chain hydrocarbons.Different crude oil samples obtained from an oil reservoir were used to isolate microorganisms for application in MEOR. Most of isolates were Pseudomonas and Bacillus strains. The growth of different microbial isolates was studied under both aerobic and anaerobic conditions at 40ºC. These isolates were found to produce extracellular biosurfactants, reduce surface tension and showed a high emulsifying activity. In addition to these capabilities, we studied the ability of these microorganisms to degrade the heavy oil fraction, in particular long-chain hydrocarbons. Several parameters were studied, such as nutritional conditions, incubation time and paraffinic composition of the mixture. Our results show that some of the isolates displayed a capacity to degrade, under aerobic and anaerobic conditions, the large alkyl chains (18+ carbons in alkyl chains) and to reduce the viscosity of hydrocarbon mixtures. Our results, show the ability of the Bacillus subtilis strains to enhance the recovery of paraffinic oil on sand pack columns. These results suggest that the microorganisms’ here isolated have interesting characteristics to be applied for MEOR

Estrutura pré- e pós-corte de capim Marandu em função da face de exposição ao sol e distância do renque em sistema silvipastoril

A avaliação estrutural do dossel é uma maneira de garantir o entendimento dos mecanismos que governam o acúmulo de forragem em sistemas de produção. O sombreamento total ou parcial de forrageiras produzidas em sistemas Silvipastoril está atrelado a uma discussão fun damental para o entendimento das respostas estruturais de gramíneas tropicais. O objetivo foi avaliar o comportamento e as características morfológicas e estruturais do capim Marandu (Brachiaria brizantha cv. Marandu), em função da face de exposição (norte e sul) e distância (3, 6, 10 e 15 metros) das arvores no sistema Silvipastoril. O delineamento foi inteiramente casualizado, em parcela subdividida (64 m²), com quatro repetições. Dosséis manejados no sistema Silvipastoril, tanto na linha quanto na face, possuem valores médios de índice de interceptação luminosa (IL), índice de área foliar (IAF) e alturas semelhantes, no pré- e no pós-corte. A altura em pós-corte foi de 12,75 cm, muito próxima da meta de resíduo (15 cm), e correspondeu a um IAF médio de 2,75 e ângulo da folhagem (ANG) de 49,5º e interceptação luminosa (IL) de 75,94%. No pré-corte os ângulos foliares corresponderam a um valor médio de 40,2º. A altura foi em média, 28 cm com IL de 95,4%. As características estruturais do pasto de Brachiaria brizantha cv. Marandu em sistema silvipastoril não foi influenciado pela face de exposição e distância do renque, enquanto as árvores têm até 9 metros de altura