Gorete Ribeiro de Macedo

Biosorption of cadmium using the fungus Aspergillus niger

Sorption experiments using the Aspergillus niger fungus for cadmium removal were carried out to study the factors influencing and optimizing the biosorption of this metal. The effects of pH, time, biomass concentration, and initial concentration of the heavy metal on the rate of metallic biosorption were examined. An experimental design was also used to determine the values of the under study variables that provided the greatest biosorption efficiency. A technique for biomass recovery was also developed with the objective of determining the capacity of the regenerated biomass to biosorb the metals in solution. This research proved that with a pH of 4.75, a biomass concentration of 0.7 g/L, and a heavy metal concentration varying between 5 and 10 mg/L a biosorption process of biosorption with Aspergillus niger could be successfully used for heavy metal removal from oil field water in the oil industry.

Production of Biosurfactant by Pseudomonas aeruginosa Grown on Cashew Apple Juice

In this work, the ability of biosurfactant production by Pseudomonas aeruginosa in batch cultivation using cashew apple juice (CAJ) and mineral media was evaluated. P. aeruginosa was cultivated in CAJ, which was supplemented with peptone (5.0 g/L) and nutritive broth. All fermentation assays were performed in Erlenmeyer flasks containing 300 mL, incubated at 30°C and 150 rpm. Cell growth (biomass and cell density), pH, and superficial tension were monitored vs time. Surface tension was reduced by 10.58 and 41% when P. aeruginosa was cultivated in nutrient broth and CAJ supplemented with peptone, respectively. These results indicated that CAJ is an adequate medium for growth and biosurfactant production. Best results of biosurfactant production were obtained when CAJ was supplemented with peptone.

Chitooligosaccharides antagonize the cytotoxic effect of glucosamine

Chitooligosaccharides (COS) are partially hydrolyzed compounds derived from chitosan that exhibit a number of biological activities, including antitumor, antibacterial and antifungal properties. In this work, we examined the cytotoxicity of pure COS and oligomers A, B and C (solutions composed of different amounts of COS) produced by enzymatic hydrolysis using a crude enzyme extract produced by the fungus Metarhrizium anisopliae. The antiproliferative effect of these molecules was analyzed using tumor cell lines (HepG2 and HeLa cells) and in a normal cell line (3T3). The antioxidant activity was analyzed in several in vitro experiments. Glucosamine showed higher toxicity (approximately 92%) to all cell lines studied. However, the oligomers obtained after hydrolysis demonstrated no toxic effects on the normal cells (3T3). Furthermore, we showed that a small amount of other COS can decrease the cytotoxic effect of glucosamine against 3T3 cells, indicating that glucosamine could be used as an antitumor drug in the presence of other COS. In addition, different effects were found in antiproliferative assays, which depended on the COS composition in the oligomers (A, B and C), showing that a combination of them may be essential for developing antineoplastic drugs. Superoxide anion scavenging was the main antioxidant activity demonstrated by the COS and oligomers. This activity was also dependent on the oligomer composition of the chitosan hydrolysates. Further work will identify the ideal proportions of COS and glucosamine for maximizing the effects of these biological activities.

Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography

Visceral leishmaniasis, a disease caused by Leishmania infantum chagasi, represents a major public health problem in many areas of the world. However, there is currently no vaccine for human use. The aim of this work was to purify the 503 antigen of Leishmania i. chagasi directly from unclarified Escherichia coli feedstock through expanded bed adsorption (EBA) chromatography. Batch experiments were performed to optimize the adsorption and elution conditions of the antigen onto a STREAMLINE Chelating resin using two central composite rotatable designs (CCRD). The results showed that the optimal binding conditions of the 503 antigen were pH 8.0 in the presence of 2.4 M NaCl. For the elution of the target protein, the optimized conditions included the presence of 600.0 mM imidazole. The adsorption isothermal data of the 503 antigen were fitted to the Langmuir adsorption isotherm. The EBA experiment successfully recovered 59.2% of the 503 antigen from the unclarified E. coli homogenate with a purification factor of 6.0.

Scientific collaboration in biotechnology: the case of the northeast region in Brazil

The objective of this study is to investigate scientific collaboration in biotechnology in the northeast region of Brazil. The data presented refer to the 1980–2010 period and were collected from the Brazilian National Council for Scientific and Technological Development platform database known as Lattes (a compilation of curricula vitae of researchers in Brazil, including a record of their scientific production) and from the Institute for Scientific Information Web of Science database. Our analysis involved the use of bibliometric indicators, specifically co-authorship between or among institutions, as well as the evaluation of social networks and multivariate statistics. Overall, we verified that collaboration takes place mostly at the intra-institutional level. At intra-regional scale, we could observe the development of four clusters in relation to the collaboration dynamics, in which geographic proximity stands out as grouping factor. At the interregional level, the partnerships revolve around institutions that count with laboratory infrastructure and research tradition in the field of biotechnology. Regarding international collaboration, it remains connected to national scientific cooperation programs.

Alcoholic fermentation of Saccharomyces cerevisiae, Pichia stipitis and Zymomonas mobilis in the presence of inhibitory compounds and seawater.

Production of cellulosic ethanol and holocellulosic ethanol from vegetable or microbial biomass starts with a hydrolysate containing compounds which may produce negative effects in the enzymatic hydrolysis and fermentation stages due to the need of pretreatment of the materials. In this way, the simultaneous presence of hydroxymethylfurfural (HMF), furfural, acetic acid, levulinic acid, and formic acid in different concentrations was tested in the fermentation using Saccharomyces cerevisiae, Pichia stipitis, and Zymomonas mobilis. The substitution of freshwater by seawater in the culture medium was also analyzed. Thus, inhibitory effects were stronger in the fermentation using P. stipitis, followed by Z. mobilis and S. cerevisiae. Formic acid and acetic acid presented more significant effects among the inhibitory compounds, followed by HMF, furfural and levulinic acid. Fermentation performed in culture medium with seawater showed promising results, especially in the ethanol yield using S. cerevisiae (0.50 g ethanol/g glucose) and Z. mobilis (0.49 g ethanol/g glucose). Whereas the production of cellulosic ethanol and holocellulosic ethanol are in early stages of development on an industrial scale, and that the availability and use of freshwater may cause socio‐environmental problems for expansion of ethanol production, the use of seawater appears as an alternative to mitigate this problem.

Chitosanase production by Paenibacillus ehimensis and its application for chitosan hydrolysis

The chitosanase production by Paenibacillus ehimensis was studied in submerged cultures and the chitosan hydrolysis was evaluated by using these enzymes without purification. The bacterium produced inducibles enzymes after 12 h of growth in a culture medium containing 0.2% (w/v) of soluble chitosan as carbon source. The enzyme production was strongly repressed by the presence of glucose. The production started as soon as the available sugars finished in the culture medium. The maximum level of chitosanase activity was 500 U.L-1 at 36°C after 36 h incubation. The crude enzyme was optimally active at pH 6.0 and 55°C and in these conditions, the enzyme presented good stability (6 days). The enzyme without purification was used to hydrolyze the chitosan which resulted chitooligosaccharides between 20 and 30 min of reaction.

Kinetic analysis of in vitro production of wild-type Spodoptera frugiperda nucleopolyhedrovirus

In this study, the kinetic behavior of Sf9 and Sf21 cells used in the production of a baculovirus biopesticide to control the pest of corn Spodoptera frugiperda was analyzed. Kinetic variables such as maximum specific growth rate, cell productivity, mean rate of infection, as well as the mean rate of occlusion body production were determined during the infection of these cell-lines with the extracellular virus of the S. frugiperda nucleopolyhedrovirus (SfMNPV). The Sf9 cell-line resulted in better viral production results (5.0 x 108 OB/mL) than the Sf21 cell-line (2.5 x 108 OB/mL).

Aplicação da metodologia de superfície de resposta no estudo da produção e extração da poligalacturonase

The aim of this work was to verify the effects of initial medium moisture content (U), addition of ammonium sulphate (N) and of potassium phosphate (P) in the production of the polygalacturonase through the solid-state fermentation, using cashew apple husk as substrate and Aspergillus niger CCT0916 as transformation agent. We also studied the best extraction conditions of the produced enzyme. The best condition of production was with U of 40%, 1% of N and 0% of P being reached an activity of the poligalacturonase of 10.1 U/g. The best extraction condition is an agitation system with a time of 100 min and a solvent-fermented medium volume ratio of 5 mL/g.

Modeling and simulation of breakthrough curves during purification of two chitosanases from Metarhizium anisopliae using ion-exchange with expanded bed adsorption chromatography

A mathematical model was developed to predict breakthrough curves during purification of the two chitosanases from Metarhizium anisopliae by expanded bed adsorption, taking into account the axial dispersion of liquid and using Streamline DEAE and SP XL adsorbents, anion and cation exchange resins, respectively. All the experiments were performed without clarification (with cells) aiming at the reduction of unit operations in future projects of separation processes, thereby reducing capital and operating costs. Chitosanases are enzymes that hydrolyze the carbohydrate chitosan, resulting in oligosaccharides that have many remarkable biological activities, such as anti-cancer, anti-HIV and antioxidant activities. The two adsorbents had similar performance in relation to hydrodynamics and mass transfer. The results of the parametric sensitivity analysis agree with the literature, and the model was validated with an average high degree of fit (94.68%) between simulated and experimental data obtained in this work.