Estudo do mecanismo de produção de oligossacarídeos com atividades nutracêuticas a partir da quitosana por hidrólise enzimática com processo fermentativo simultâneo

The obtaining of the oligosaccharides from chitosanase, has showed interest of the pharmaceutical area in the last years due their countless functional properties. Although, the great challenge founded out is how to keep a constant and efficient production. The alternative proposed by this present work was to study the viability to develop an integrated technology, with reduced costs. The strategy used was the obtaining of the oligomers through enzymatic hydrolysis using chitosanolitic enzymes obtained straight from the fermented broth, eliminating this way the phases involved in the enzymes purification. The two chitosanases producing strains chosen for the work, Paenibacillus chitinolyticus and Paenibacillus ehimensis, were evaluated according to the behavior in the culture medium with simple sugar and in relation to the pH medium variations. The culture medium for the chitosanases induction and production was developed through addition of soluble chitosan as carbon source. The soluble chitosan was obtained using hydrochloric acid solution 0.1 M and afterwards neutralization with NaOH 10 M. The enzymatic complexes were obtained from induction process in culture medium with 0.2% of soluble chitosan. The enzymes production was verified soon after the consumption of the simple sugars by the microorganisms and the maximum chitosanolitic activity obtained in the fermented broth by Paenibacillus chitinolyticus was 249 U.L-1 and by Paenibacillus ehimensis was 495U.L-1. These two enzymatic complexes showed stability when stored at 20°C for about 91 days. The enzymes in the fermented broth by Paenibacillus chitinolyticus, when exposed at temperature of 55°C and pH 6.0, where the activity is maximum, showed 50% lost of activity after 3 hours Meanwhile, for the complex produced by Paenibacillus ehimensis, after 6 days of exposure, it was detected 100% of the activity. The chito-oligosaccharides obtained by the hydrolysis of a 1% chitosan solution, using the enzymatic complex produced by Paenibacillus chitinolyticus showed larger quantity after 9 hours hydrolysis and using the complex produced by Paenibacillus ehimensis after 20 minutes was observed the chito-ligosacharides with polymerization degree between 3 and 6 units. Evaluating these results, it was verified that the production of chitosan-oligosaccharides is possible, using a simultaneous process

Recuperação e purificação de proteínas do soro de queijo tipo coalho usando cromatografia de troca iônica e interação hidrofóbica em leito na forma expandida

Expanded Bed Adsorption plays an important role in the downstream processing mainly for reducing costs as well as steps besides could handling cells homogenates or fermentation broth. In this work Expanded Bed Adsorption was used to recover and purify whey proteins from coalho cheese manufacture using Streamline DEAE and Streamline SP both ionic resins as well as a hydrophobic resin Streamline Phenyl. A column of 2.6 cm inner diameter with 30 cm in height was coupled to a peristaltic pump. Hydrodynamics study was carried out with the three resins using Tris-HCl buffer in concentration of 30, 50 and 70 mM, with pH ranging from 7.0 to 8.0. In this case, assays of the expansion degree as well as Residence Time Distribution (RTD) were carried out. For the recovery and purification steps, a whey sample of 200 mL, was submitted to a column with 25mL of resin previously equilibrated with Tris/HCl (50 mM, pH 7.0) using a expanded bed. After washing, elution was carried out according the technique used. For ionic adsorption elution was carried out using 100 mL of Tris/HCl (50 mM, pH 7.0 in 1M NaCl). For Hydrophobyc interaction elution was carried out using Tris/HCl (50 mM, pH 7.0). Adsorption runs were carried out using the three resins as well as theirs combination. Results showed that for hydrodynamics studies a linear fit was observed for the three resins with a correlation coefficient (R2) about 0.9. In this case, Streamline Phenyl showed highest expansion degree reaching an expansion degree (H0/H) of 2.2. Bed porosity was of 0.7 when both resins Streamline DEAE and Streamline SP were used with StremLine Phenyl showing the highest bed porosity about 0.75. The number of theorical plates were 109, 41.5 and 17.8 and the axial dipersion coefficient (Daxial) were 0.5, 1.4 and 3.7 x 10-6 m2/s, for Streamline DEAE, Streamline SP and Streamline Phenyl, respectively. Whey proteins were adsorved fastly for the three resins with equilibrium reached in 10 minutes. Breakthrough curves showed that most of proteins stays in flowthrough as well as washing steps with 84, 77 and 96%, for Streamline DEAE, Streamline SP and Streamline Phenyl, respectively. It was observed protein peaks during elution for the three resins used. According to these peaks were identified 6 protein bands that could probably be albumin (69 KDa), lactoferrin (76 KDa), lactoperoxidase (89 KDa), β-lactoglobulin (18,3 KDa) e α-lactoalbumin (14 KDa), as well as the dimer of beta-lactoglobulin. The combined system compound for the elution of Streamline DEAE applied to the Streamline SP showed the best purification of whey proteins, mainly of the α-lactoalbumina

Recuperação e purificação de ramnolipídeos produzidos por pseudomonas aeruginosa P029-GVIIA utilizando melaço de cana como substrato

Biosurfactants are molecules produced by microorganisms mainly bacteria as Pseudomonas and Bacillus. Among the biosurfactants, rhamnolipids play an important role due to their tensoactive as well as emulsifying properties. Besides can be produced in a well consolidated way the production costs of biosurfactants are quite expansive mainly if downstream processing is goning to be considered. Actually, attention has been given to identification of biosurfactants as well as optimization of its fermentative processes including downstream ones. This work deals with the development of strategies to recovery and purification of rhamnolipids produced by Pseudomonas aeruginosa P029-GVIIA using sugar-cane molasses as substrate. Broth free of cells was used in order to investigate the best strategies to recovery and purification produced by this system. Between the studied acids (HCl and H2SO4) for the acid precipitation step, HCl was the best one as has been showed by the experimental design 24. Extraction has been carried out using petroleum ether and quantification has been done using the thioglycolic acid method. Adsorption studies were carried out with activated carbon in a batch mode using a 24 experimental design as well as combined with an hydrophobic resin Streamline Phenyl aiming to separate the produced biosurfactant. Biosurfactant partial identification was carried out using High Performance Liquid Chromatography (HPLC). Experiments in batch mode showed that adsorption has been controlled mainly by pH and temperature. It was observed a reduction of 41.4% for the liquid phase and the solid phase it was possible to adsorb up to 15 mg of rhamnolipd/g of activated carbon. The kinetics of adsorption has been well fitted to a pseudo-first order reaction with velocity constant (k1) of 1.93 x 10-2 min-1. Experiments in packed bed ranging concentration on eluent (acetone) has been shown the highest recovery factor of 98% when pure acetone has been used. The combined effect if using activated carbon with an hydrophobic resin Streamline Phenyl has been shown successful for the rhamnolipids purification. It has been possible to purify a fraction of the crude broth with 98% of purity when the eluted of activated carbon packed bed was used with pure acetone

Avaliação da Qualidade da Água do rio Pitimbu

The Pitimbu River Basin is concern reason, therefore this is one of the mains freshwater sources for the Natal city supplying. The Pitimbu waters river flow into Jiqui pond, as the main supplier of this, that supplies 16% of Natal population and there is a bigger importance despite the supplying of a parcel of Natal city made joining underground waters with the water proceeding from the Jiqui pond, being used for dilution waters of contaminated wells with nitrate. Even with the importance of the Pitimbu, there is a strong pressure of occupation of its edge according to a critical urban growth, becoming the situation worse and increasing the necessity of management of the use and occupation of the ground and keeping the control of the prompt pollution, as punctual as diffuse. There are many studies about Pitimbu River Basin, however they summarize themselves to some months or even in a few years. This work, that gathers these information, increasing the amplitude and making possible an evaluation of the attitude of the water in a period extended for an evaluation of values of variable quality of water referring to the standards, that usually happens about the variable frequency distribution, central tendency and measures of dispersion, as the coefficient of variation, coefficient of skewness and the coefficient of determination of certain variable on another one. This work to evaluates the quality conditions of the Pitimbu river waters by analyzing thirty five (35) variables Physical, chemical and biological in eight (8) points of water course since 1993 to 2007. Given this situation, all knowledge about the waters quality conditions obtained in this work, is a strong subsidy for management of use and ground occupation, considering the river basin as territorial unit of management, and as the water is public good domain, being a priority human use, It is necessary to guarantee to current and the future generations available water resources in appropriate standards of quality as the established standard and identifying possible reasons of pollution through statistics analysis

New Culture Medium to Xanthan Production by Xanthomonas campestris pv. campestris

Xanthan is a important biopolymer for commercial purpose and it is produced in two stages by Xanthomonas campestris. In the first one, the bacterium is cultivated in the complex medium enriched in nitrogen and the biomass produced is used as inoculum for the next stage in which the gum is produced in another medium. In this work a new medium for the first stage is proposed in place of currently used YM medium. Different formulated growth media were studied and the correspondent biomass produced was analysed as inoculum for the second stage. The inoculum and gum were produced by batch process in shaker at 27A degrees C in pH 6.0 and at 30A degrees C in pH 7.0, respectively. The gum was precipitated with ethanol (3:1 v/v). The dryed biomass and xathan gum produced were determined by drying in oven at 105 and 40A degrees C, respectively. The viscosity of the fermentation broth and 1% gum solution in water were determined in Brookfield viscometer. The formulated medium presented the increase in gum production (30%), broth (136%) and 1% gum solution viscosity (60%) compared to YM, besides the inferior cost. The results showed the importance of the quality of the inoculum from the first stage of the culture which influenced on the gum viscosity in the second stage.

Susceptibility of Saccharomyces cerevisiae and lactic acid bacteria from the alcohol industry to several antimicrobial compounds

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Surface-enhanced resonance Raman scattering: Single-molecule detection in a Langmuir-Blodgett monolayer

Surface-enhanced resonance Raman scattering (SERRS) is used for single-molecule detection from spatially resolved 1-mum(2) sections of a Langmuir-Blodgett (LB) monolayer deposited onto a Ag film. The target molecule, his (benzimidazo) thioperylene (BZP), is dispersed in an arachidic acid monomolecular layer containing one BZP molecule per mum(2) which is also the probing area of the Raman microscope. For concentrated samples (attomole quantities in the field of view), average SERRS, surface-enhanced fluorescence (SEF), and Raman imaging, including line mapping and global images at different temperatures, were recorded. Single-molecule SERRS spectra, obtained using an LB monolayer, present changes in bandwidth and relative intensities, highlighting the properties of single-molecule SERRS that are lost in average SERRS measurements of mixed LB monolayers obtained at the same temperatures. Also, the dilute system phenomenon of blinking is discussed with regard to results obtained from LB monolayers. The dilution process used in the single-molecule LB SERRS work is independently supported by fluorescence results obtained from very dilute solutions with monomer concentrations down to 10(-12) M.

A Nonstaining and Tasteless Hydrophobic Salt of Chlorhexidine

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effect of Usnic Acid on Candida orthopsilosis and C. parapsilosis

The activity of usnic acid against Candida orthopsilosis and Candida parapsilosis on planktonic and biofilm conditions was investigated by using a broth microdilution and microplate methods. Potent in vitro activities against different Candida species were obtained. The metabolic activity of sessile cells of C. parapsilosis complex was reduced by 80% at four times the 80% inhibitory concentration. The in vitro studies support further efforts to determine whether usnic acid can be used clinically to cure patients with Candida infections.

Anti-Helicobacter pylori activity and immunostimulatory effect of extracts from Byrsonima crassa Nied. (Malpighiaceae)

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

Anti-Mycobacterium tuberculosis activity of fungus Phomopsis stipata

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

Rapid detection of resistance to pyrazinamide in Mycobacterium tuberculosis using the resazurin microtitre assay

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Memórias midiatizadas: percepções sobre ciência reconfiguradas a partir do consumo midiático

This piece focuses on the issue of media in the contemporary context, taking for its object the sociocultural phenomenon of mediatization‟, understood as the dilution of media culture in the everyday life, according to the perspective from Fausto Neto (2008) e Sodré (2002). This phenomenon is viewed from the angle of reception, that is, one that considers the individuals as pro-actives in this dynamic. The media culture is perceived as a fundamental element in the process of social structuring, and also is taken as a symbolic arm of great importance for the elaboration of the notion of reality as it interferes with the flow of information. Thus, we emphasize the process of symbolic consumption that we do with media content (Canclini, 1999), through a dynamic of appropriation and reframing, given from the interweaving of the meanings proposed with extra-media elements, recaptured from the perspective of mediations, by Barbero (2006), which concerns the articulation of the contents we appropriate from media with interactions from other social practices. We also consider the dimension of memory according to Sarlo (2007) and Halbwachs (2004) in order to, through speeches made in its scope, reach empirically the phenomenon of mediatization‟, and overlapping it, the media consumption on a specific theme, "science". Nevertheless, in terms of field work, we use the technique of in-depth interviews, so that the speech of our interviewers was our corpus. Through their narratives, speeches considered by us as memory-related, we visualize the dynamics of media consumption on given topic. At the level of conclusions, we realize that mediatization‟ as a phenomenon occurred, but its flows and orders showed discontinuities and pluralities not initially conceived; regarding the consumption of science through media, we see hybrid perceptions by our respondents, that is, one that include both elements aligned with the positivist conception of science - which we consider to be the view of science proposed by the media and elements that would contradict this view

Os argumentos transcendentais: kant e o problema de hume

This work whose title is "The transcendental arguments: Kant Andy Hume's problem" has as its main objective to interpret Kant's answer to Hume's problem in the light of the conjunction of the causality and induction themes which is equivalent to skeptical- naturalist reading of the latter. In this sense, this initiative complements the previous treatment seen in our dissertation, where the same issue had been discussed from a merely skeptical reading that Kant got from Hume thought and was only examined causality. Among the specific objectives, we list the following: a) critical philosophy fulfills three basic functions, a founding, one negative and one would argue that the practical use of reason, here named as defensive b) the Kantian solution of Hume's problem in the first critisism would fulfill its founding and negative functions of critique of reason; c) the Kantian treatment of the theme of induction in other criticisms would will fulfill the defense function of critique of reason; d) that the evidence of Kant's answer to Hume's problem are more consistent when will be satisfied these three functions or moments of criticism. The basic structure of the work consists of three parts: the first the genesis of Hume's problem - our intention is to reconstruct Hume's problem, analyzing it from the perspective of two definitions of cause, where the dilution of the first definition in the second match the reduction of psychological knowledge to the probability of following the called naturalization of causal relations; whereas in the second - Legality and Causality - it is stated that when considering Hume in the skeptic-naturalist option, Kant is not entitled to respond by transcendental argument A􀁴B; A⊢B from the second Analogy, evidence that is rooted in the position of contemporary thinkers, such as Strawson and Allison; in third part - Purpose and Induction - admits that Kant responds to Hume on the level of regulative reason use, although the development of this test exceeds the limits of the founding function of criticism. And this is articulated in both the Introduction and Concluding Remarks by meeting the defensive [and negative] function of criticism. In this context, based on the use of so-called transcendental arguments that project throughout the critical trilogy, we provide solution to a recurring issue that recurs at several points in our submission and concerning to the "existence and / or the necessity of empirical causal laws. In this light, our thesis is that transcendental arguments are only an apodictic solution to the Hume s skeptical-naturalist problem when is at stake a practical project in which the interest of reason is ensured, as will, in short, proved in our final considerations

Monitoring of the dissemination of Salmonella in the chicken Frankfurt-sausage production line of a sausage factory in the state of São Paulo, Brazil

Poultry meat and its derivatives are among the foodstuffs considered by environmental health authorities to present the highest risks to the public. A total of 185 samples were collected in five monthly batches, from different processing stages in a sausage plant that uses mechanically-deboned chicken meat (MDCM) and tested for the presence of Salmonella. Enrichment was carried out in both Kauffman tetrathionate broth and Rappaport-Vassiliadis broth and isolation on Salmonella-Shigella agar and brilliant-green agar. Live Salmonella bacteria were isolated from six samples of the raw meat and from the emulsion, in batches three, four and five, but not from any sample in batches one or two. The six isolated strains were all classified as Salmonella Albany, which has not previously been reported in MDCM. of the two enrichment broths, Rappaport-Vassiliadis gave the better results. The Pattern of contamination suggests a probable common source, given that a new supplier was used in the third, fourth, and fifth months. It was also shown that the industrial cooking was effective in preventing Salmonella surviving in the final product.