EXPANDED BED ADSORPTION OF BROMELAIN (EC 3.4.22.33) FROM Ananas comosus CRUDE EXTRACT

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)This work focuses on the adsorption of Bromelain in expanded bed conditions, such as the adsorption kinetics parameters. The adsorption kinetics parameters showed that after 40 minutes equilibrium was achieved and maximum adsorption capacity was 6.11 U per resin mL. However, the maximum adsorption capacity was only determined by measuring the adsorption isotherm. Only by the Langmuir model the maximum adsorption capacity, Qm, and dissociation constant, kd, values could be estimated as 9.18 U/mL and 0.591, respectively, at 25 degrees C and 0.1 mol/L phosphate buffer pH 7.5. A column made of glass with an inner diameter of 1 cm was used for the expanded bed adsorption (EBA). The residence time was reduced 10 fold by increasing the expansion degree 2.5 times; nonetheless, the plate number (N) value was reduced only 2 fold. After adsorption, the bromelain was eluted in packed bed mode, with a downward flow. The purification factor was about 13 fold and the total protein was reduced 4 fold. EBA showed to be feasible for purification of bromelain.261149157Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Partitioning optimization of proteins from Zea mays malt in ATPS PEG 6000/CaCl2

This work aimed to establish the relationship between the compositions and pH of ATPS PEG 6000/CaCl2 and the proteins partition from maize malt and also to simplify the process optimization in ATPS for a statistical model, established by response surface methodology (RSM). Results showed that these were no influence of pH on the phase diagrams and on the composition of tie line length of PEG 6000/CaCl2 ATPS. SRM analyses showed that elevated pH and larger tie line length were the best conditions for recovering of maize malt proteins. The maximum partition coefficient by PEG 6000/CaCl2 ATPS was about 4.2 and was achieved in ATPS in a single purification step. The theoretical maximum partition coefficient was between 4.1-4.3. The process was very suitable for continuous aqueous two phase purification due to the stability of proteins (e.g. alpha and beta-amylases) and could increase their content into middle.50355756

A Mycobacterium leprae Hsp65 Mutant as a Candidate for Mitigating Lupus Aggravation in Mice

Hsp60 is an abundant and highly conserved family of intracellular molecules. Increased levels of this family of proteins have been observed in the extracellular compartment in chronic inflammation. Administration of M. leprae Hsp65 [WT] in [NZBxNZW]F1 mice accelerates the Systemic Lupus Erythematosus [SLE] progression whereas the point mutated K409A Hsp65 protein delays the disease. Here, the biological effects of M. leprae Hsp65 Leader pep and K409A pep synthetic peptides, which cover residues 352–371, are presented. Peptides had immunomodulatory effects similar to that observed with their respective proteins on survival and the combined administration of K409A+Leader pep or K409A pep+WT showed that the mutant forms were able to inhibit the deleterious effect of WT on mortality, indicating the neutralizing potential of the mutant molecules in SLE progression. Molecular modeling showed that replacing Lysine by Alanine affects the electrostatic potential of the 352–371 region. The number of interactions observed for WT is much higher than for Hsp65 K409A and mouse Hsp60. The immunomodulatory effects of the point-mutated protein and peptide occurred regardless of the catalytic activity. These findings may be related to the lack of effect on survival when F1 mice were inoculated with Hsp60 or K409A pep. Our findings indicate the use of point-mutated Hsp65 molecules, such as the K409A protein and its corresponding peptide, that may minimize or delay the onset of SLE, representing a new approach to the treatment of autoimmune diseases

Premolis semirufa (Walker, 1856) Envenomation, Disease Affecting Rubber Tappers of the Amazon: Searching for Caterpillar-Bristles Toxic Components

Pararama, the popular name of the larval form of the moth Premolis semirufa inhabits rubber plantations in the Amazon region and the accidental contact of the skin with the caterpillar's bristles or cocoons results in immediate and intense heat, pain, edema, and itching. In many cases a chronic inflammatory reaction with immobilization of the joints occurs. The current study has evaluated the biological and immunochemical characteristics of the Pararama caterpillar bristles extract. Electrophoretic analysis showed the presence of several components, including a very intense 82 kDa band. This latter component was endowed with intense gelatinolytic activity, as observed in zymography assays. Further analysis revealed that the extract also contained hyaluronidase activity but is devoid of phospholipase A2 activity. In vivo assays, using mice, showed that the extract was not lethal, but caused significant edema and induced intense infiltration of inflammatory cells to the envenomation site. The extract also induced high specific antibody titers, but no autoantibodies were detected. The data obtained, so far, demonstrate the existence of a mixture of different enzymes in the bristles of Premolis semirufa caterpillar, which can act together in the generation and development of the clinical manifestations of the Pararama envenomation

Continuous extraction of xylanase from Penicillium janthinellum with reversed micelles using experimental design mathematical model

Xylanase recovery from Penicillium janthinellum with a reversed micellar system consisting of a cationic surfactant using continuous process was evaluated. A statistical approach was applied to the results and showed that the highest xylanase recovery (140%), indicated by the model, was attained at an ionic strength of 7.5 mS cm(-1) and volumetric flow of 0.6 ml min(-1). A new xylanase extraction was performed under these conditions to test the model and showed a close similarity between the experimental result and the value predicted by the model.23536536

Production of xylanase and protease by Penicillium janthinellum CRC 87M-115 from different agricultural wastes

Five agricultural wastes were evaluated in submerged fermentation for xylanolytic enzymes production by Penicillium janthinellum. The wastes were hydrolyzed in acid medium and the liquid fraction was used for cultivation. Corn cob (55.3 U/mL) and oat husk (54.8 U/mL) were the best inducers of xylanase. Sugar cane bagasse (23.0 U/mL) and corn husk (23.8 U/mL) were moderately good, while cassava peel was negligible. Protease production was very low in all agro-industrial residues. The maximum biomass yields were 1.30 and 1.17 g/L for cassava peel and corn husk after 180 h, respectively. Xylanolytic activity showed a cell growth associated profile. (c) 2005 Elsevier Ltd. All rights reserved.97686286

Inorganic polyphosphate accumulation by Cunninghamella elegans (UCP 542) and its influence in the decolorization of textile azo dye Orange II

The inappropriate disposal of dyes in wastewater constitutes an environmental problem and can cause damage to the ecosystem. Alternative treatments have been reported that employ fungi that are particularly effective in the decolorization of textile effluents. In these methods, the biomass can be used for industrial removal of dyes by biosorption. The current study has investigated the accumulation of polyphosphate (poly P) in the fungus Cunninghamella elegans UCP 542 and evaluated the capacity of the inactivated biomass, obtained from 144 h of incubation, to decolorize the dye Orange II. The results suggest the transport of the inorganic phosphorus, as demonstrated through the cellular bioaccumulation. The biochemical and physiological aspects of bioaccumulation in the poly P form were related to the process of removal of the color of Orange II, with the reduction of 60 % of the dye solution.15117918

Neural modeling for cytochrome b5 extraction

In this work a neural model for cytochrome b5 extraction in batch and in continuous operation was developed. The best feedforward arquiteture achieved for batch operation modeling was 3-4-1 and 3-8-2 for the continuous operation. It was observed that among the models developed, the best adjustment was that obtained with Bayesian regularization algorithm training. Deviations of less than 10% were observed for the experimental data and they are similar for the neural model, since it was statistically proved the null hypothesis in the comparison between the two independent samples (experimental and predicted outputs). (c) 2006 Elsevier Ltd. All rights reserved.4161272127