Produção de biossurfactantes: manoproteínas intracelulares e soforolipídios extracelulares por Saccharomyces cerevisiae / Biosurfactant production: intracellular manoproteins and extracellular sophorolipids by Saccharomyces cerevisiae

RESUMO Os biossurfactantes apresentam inúmeras aplicações ambientais e são produzidos por diversos microrganismos. Os provenientes da levedura Saccharomyces cerevisiae são pouco estudados para fins ambientais, sendo atóxicos. Objetivou-se o estudo da produção de biossurfactantes intra e extracelular por essa levedura, desenvolvida em meio de cultivo contendo 0,5% de extrato de levedura e 1% de peptona, além de concentrações variadas de sacarose e indutores oleosos - glicerol e óleos de soja e diesel. Os experimentos foram realizados durante 96 horas, e a produção de biossurfactantes foi avaliada diariamente, por meio da redução da tensão superficial e de estabilização de emulsões. O biossurfactante extracelular foi extraído da biomassa obtida, com posterior precipitação e caracterização química por intermédio de espectrometria de massa. As maiores produtividades de emulsificantes extracelulares foram obtidas com glicerol (0,20 UE.h-1) e óleo de soja (0,21 UE.h-1), em 48 horas de cultivo. Em ensaios posteriores, realizados com aumento da concentração de indutor, foi verificado um aumento das produtividades extracelulares para 0,45 UE.h-1 para o glicerol e 0,30 UE.h-1 para o óleo de soja. A maior redução da tensão superficial foi de 9,89%, em 72 horas, para o indutor óleo diesel. A diminuição dessa tensão, aliada ao aumento das atividades emulsificantes, é um importante indicativo da utilização do substrato hidrofóbico pelo microrganismo. O estudo comprova aumento na produção de biossurfactantes extracelulares quando realizada otimização de cultivo. Para a produção dos intracelulares, a necessidade de processo de rompimento celular aumenta os custos do bioprocesso.

ABSTRACT Biosurfactants implicate many environmental applications, being produced by a wide range of microorganisms. Those from the Saccharomyces cerevisiae yeast are still poorly studied for environmental purposes and are non-toxic. The aim of the study was the production of intra- and extracellular biosurfactants by the Saccharomyces cerevisiae yeast. The yeast was grown in cultured medium containing 0.5% yeast extract, 1% peptone and variable concentrations of sucrose and oily inducers. Inducers used were glycerol, soybean oil and diesel oil. Experiments were conducted for 96 h, and the daily production of biosurfactants was evaluated by reducing surface tension and stabilizing emulsions. Extracellular biosurfactant was extracted from the obtained biomass, with subsequent precipitation and chemical characterization by mass spectrometry. The highest extracellular emulsifier yields were achieved with glycerol inductor (0.20 UE h-1) and soybean oil (0.21 UE h-1) in 48h of cultivation. In later tests performed with increasing concentration of inducer, an increase in extracellular yields was noticed in these experiments (0.45 UE h-1 for glycerol and 0.30 UE h-1 for the soybean oil). The greatest reduction in surface tension was 9.89% in 72 h for diesel oil inducer. The reduction of surface tension combines with the increase of emulsifying activities in an important indicator of the use of hydrophobic substrate by the microorganism. The study confirms an increase in the production of extracellular biosurfactants when optimizing cultivation. The production of intracellular biosurfactants has also been verified, however the process of cellular disruption increases the cost of the bioprocess.

Pilarização de argilas e perspectivas de aplicação e de pesquisa agronômica e ambiental / Pillaring of clays and perspective of agronomic and environmental application and research

A exploração das propriedades químicas e estruturais de argilas expansivas, por meio da intercalação de polímeros inorgânicos no espaço interlamelar, permite o desenvolvimento de materiais reativos e com elevado potencial de aplicação tecnológica, agronômica e ambiental. Esta técnica, conhecida como pilarização, já é bastante difundida na área das ciências química e dos materiais, havendo esforços crescentes nos últimos anos para aplicação desses materiais com fins ambientais. Contudo, apenas uma fração do conhecimento acumulado é atualmente utilizada diretamente por cientistas do solo, abrindo oportunidades para novas pesquisas por meio do intercambio de conhecimento entre as diferentes áreas de pesquisa, como físico-química e microbiologia. Portanto, esta revisão apresenta e discute o que são a pilarização e as argilas pilarizadas, métodos de síntese e algumas aplicações nas ciências do solo e ambiental, buscando encorajar mais grupos brasileiros de pesquisa a explorá-las.

The use of chemical and structural properties of swelling clays intercalated with inorganic polymers allows the development of materials with high reactivity and potential for technological and environmental applications. This technique is known as pillaring and has been studied widely in the field of chemical and material science, but the use of pillared clays for environmental application has increased only in the last few years. Nevertheless, only a fraction of this knowledge is applied by soil scientists being, therefore, an opportunity to develop new studies including a wide range of disciplines, such as physico-chemistry and microbiology. Therefore, this review presents and discusses what are pillaring and pillared clays, methods of synthesis and some applications in soil and environmental science, aiming to encourage more research groups in Brazil to explore it.

Production of heterologous cutinases by E. coli and improved enzyme formulation for application on plastic degradation

Background: The hydrolytic action of cutinases has been applied to the degradation of plastics. Polyethylene terephthalate (PET) have long half-life which constitutes a major problem for their treatment as urban solid residues. The aim of this work was to characterize and to improve stable the enzyme to optimize the process of degradation using enzymatic hydrolysis of PET by recombinant cutinases. Results: The wild type form of cutinase from Fusarium solani pisi and its C-terminal fusion to cellulose binding domain N1 from Cellulomonas fimi were produced by genetically modified Escherichia coli. The maximum activity of cutinases produced in Lactose Broth in the presence of ampicillin and isopropyl β-D-1-thiogalactopyranoside (IPTG) was 1.4 IU/mL. Both cutinases had an optimum pH around 7.0 and they were stable between 30 and 50ºC during 90 min. The addition of glycerol, PEG-200 and (NH4)2SO4 to the metabolic liquid, concentrated by ultra filtration, stabilized the activity during 60 days at 28ºC. The treatment of PET with cutinases during 48 hrs led to maxima weight loss of 0.90%. Conclusions: Recombinant microbial cutinases may present advantages in the treatment of poly(ethylene terephthalate) PET through enzymatic treatments.