Purification processes and market potential of bromelain in Brazil

Bromelain is the denomination given to the group of endoproteases obtained from members of Bromeliacea family. These enzymes have a wide range of proven applications and have been an object of study for worldwide researchers for decades. Over the years, several different downstream processes were studied in order to determine which technique would be worthwhile to be implemented in Brazil and provide the national market with such product. The objective of the present study is to relate the main studies in Brazil that has proven that bromelain purification can be cost-effective, in addition to the well-known benefits owned by such enzymes, and highlight the applications that create their market potential in the Brazilian market8882888CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçãoThe authors would like to acknowledge the financial support of FAPESP (São Paulo Research Fundation), PROPP-UFU (Dean of Research and Graduate Studies at the Federal University of Uberlândia) and CNPq (National Council for Scientific and Technological Development

The effect of pH on bromelain partition from Ananas comosus by PEG4000/phosphate ATPS

In this work, the thermodynamic equilibrium and applying of PEG4000/Phosphate ATPS on the purification of bromelain extracted from pineapple was studied. A rigorous study of the equilibrium curves and tie-line length from PEG4000/phosphate ATPS were done for the pH 6-11 at 25ºC. Results showed that there was augment in the PEG and salt contents with the high pH value from PEG4000/Phosphate ATPS and two-phase formation needed only increasing the PEG content. Two tie-line length at pH 11 from PEG4000/Phosphate ATPS were optimal condition for bromelain purification, one on composition of 14% PEG and 13% salt and other at 12.6% PEG and 12.2% salt, while a 25-62 folds of enzyme was found. SDS-PAGE electrophoreses had one band only, which showed that bromelain was purified. Optimum conditions of bromelain use were found at pH 7 and between 30-40ºC.125132Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Transporte de particulas em leitos fluidizados

Orientador: João Alexandre F. Rocha PereiraDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de CampinasResumo: O trabalho descrito na presente tese tem como primeiro objetivo correlacionar o transporte de partículas em leitos fluidizados quando são utilizadas elevadas velocidades de gás. Estuda-se o efeito da velocidade do gás e do espaço de separação ("FREEBOARD") na taxa de partículas transportadas. Os resultados obtidos nos permitem distinguir elutriação do transporte por ejeção de partículas na região de separação. Estes resultados mostram que, a baixas velocidades de fluidização, as partículas transportadas seguem uma lei de distribuição do tipo gama. Em altas velocidades de fluidização, as partículas transportadas seguem a mesma distribuição do leito de partículas, aproximadamente uma distribuição do tipo normal truncada ¿Observação: O resumo, na íntegra poderá ser visualizado no texto completo da tese digital.Abstract: The work described in the present thesis first attempts to correlate the transport of particles in fluidized beds when high gas velocities are used. The effect of gas velocities and Freeboard height upon rate of particle transport is studied. The results permit to distinguish between elutriation and transport due to ejection of particles in the Freeboard region. It was found that the transported particles follows the gamma distribution at low gas velocities, and at high gas velocities it follows the same distributions as the bed particles, approximately the truncated Gauss distribution ...Note: The complete abstract is available with the full electronic digital thesis or dissertations.MestradoMestre em Engenharia Químic

Sphingomyelinase D Activity in Sicarius tropicus Venom : Toxic Potential and Clues to the Evolution of SMases D in the Sicariidae Family

The spider family Sicariidae includes three genera, Hexophthalma, Sicarius and Loxosceles. The three genera share a common characteristic in their venoms: the presence of Sphingomyelinases D (SMase D). SMases D are considered the toxins that cause the main pathological effects of the Loxosceles venom, that is, those responsible for the development of loxoscelism. Some studies have shown that Sicarius spiders have less or undetectable SMase D activity in their venoms, when compared to Hexophthalma. In contrast, our group has shown that Sicarius ornatus, a Brazilian species, has active SMase D and toxic potential to envenomation. However, few species of Sicarius have been characterized for their toxic potential. In order to contribute to a better understanding about the toxicity of Sicarius venoms, the aim of this study was to characterize the toxic properties of male and female venoms from Sicarius tropicus and compare them with that from Loxosceles laeta, one of the most toxic Loxosceles venoms. We show here that S. tropicus venom presents active SMases D. However, regarding hemolysis development, it seems that these toxins in this species present different molecular mechanisms of action than that described for Loxosceles venoms, whereas it is similar to those present in bacteria containing SMase D. Besides, our results also suggest that, in addition to the interspecific differences, intraspecific variations in the venoms' composition may play a role in the toxic potential of venoms from Sicarius species.Peer reviewe

Purification of bromelain enzyme from Curauá (Ananaserectifolius LB Smith) white variety, by aqueous two-phase system PEG 4000/potassium phosphate

The Curauá (Ananaserectifolius LB Smith) is a species belonging to the family Bromeliaceae. Occurs in the states of Para, Amazonas, Amapá, Acre, Mato Grosso and Goiás, has two varieties, purple and white, and its fibers are used in automotive and textile industries due to its strength, softness and lightweight. Currently, only the fiber is used in industry, the rest is considered waste. However, this residue contains compounds with important properties to be discovered and studied. An enzyme complex found in this residue is bromelain, a group of proteolysis’ enzymes with application in several areas such as in the food, pharmaceutical and cosmetics industries. Today, its use is aiming the pharmaceutical industry, the production of ointments, gels, creams and lotions because they offer a wide range of therapeutic efficacies: antiedemas, anti-inflammatory, antithrombotic and fibrinolytic activities. This work studied the enzyme purification and recovery presented in the leaves of Curauá by means of an ATPS (aqueous two-phase system) PEG 4000/potassium phosphate. The protein content was measured by Bradford reagent and the enzymatic activity was measured by the Biuret reagent. Batch assays were performed aiming the enzyme extraction and recuperation, using the partition coefficient as indicator. It was used pH 7.0, 8.0 and 9.0, varying the proportional composition between the polymeric and the saline phases (tie-lines). The white variety of Curauá was used. The best purification factor was 4.53 and about 113.04 µg/mL of total protein measured by Bradford8395399CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçã

Comparison of the C-mediating killing activity and C-activating properties of mouse monoclonal and polyclonal antibodies against Trypanosoma cruzi

A Mouse polyclonal antiserum against Trypanosoma cruzi or its IgG and IgM fractions and five monoclonal antibodies (two IgM, two IgG1 and one IgG2a) recognize and combine with membrane components of trypomastigote forms of the parasite as revealed by immunofluorescence. Although all these antibodies sensitize trypomastigotes and prepare them to activate the complement (C) system, as measured by consumption of total C, C4, B and C3, only the polyclonal antiserum or its IgG, IgM and Fabμ fragments were able to induce trypanosome lysis by the alternative C pathway

Isolation and purification of bromelain from waste peel of pineapple for therapeutic application

The aim of this work was to isolate and purify bromelain extracted from the pineapple peel by ammonium sulfate precipitation (40-80%), followed by desalting and freeze-drying with a 75% activity recovery and 2.2 fold increased specific activity. Ion exchange chromatography on DEAE-Sepharose was able to separate the polysaccharides from the enzyme, which was recovered in the elution step, maintaining its enzymatic activity. The batch adsorption of bromelain was evaluated in terms of total protein and enzymatic activity using Langmuir and Langmuir-Freundlich models. Results showed that the process could be suitable for the recovery and purification of the enzyme, maintaining its specific activity.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Estadual de Campinas Faculdade de Engenharia QuímicaUniversidade Federal de São Paulo (UNIFESP) Departamento de Ciências Exatas e da TerraUniversidade Federal de Uberlândia Instituto de Genética e BioquímicaUniversidade Estadual de Campinas Faculdade de Ciências MédicasUNIFESP, Depto. de Ciências Exatas e da TerraSciEL

Dilute-phase pneumatic conveying of polystyrene particles: pressure drop curve and particle distribution over the pipe cross-section

During the pneumatic conveying of plastic pellets, it has been observed that materials with similar physical characteristics may develop a substantial difference in pressure drop. In this work, the pressure drop in a particle-laden 2.7 meter long horizontal channel with circular cross-section is presented from an experimental perspective. Experiments are carried out for cylindrical polystyrene beads with an average diameter of 3.2 mm and mass loadings of 0.06 to 0.11 (kg particles/kg gas). The air mass flow rate was studied in the range from 0.085 kg/s to 0.170 kg/s. The pressure drop curve is shown as a function of air velocity and particle load. Response surface methodology showed high statistical significance for air velocity, particle load and their cross-relation.8188Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Isolation and purification of bromelain from waste peel of pineapple for therapeutic application

The aim of this work was to isolate and purify bromelain extracted from the pineapple peel by ammonium sulfate precipitation (40-80%), followed by desalting and freeze-drying with a 75% activity recovery and 2.2 fold increased specific activity. Ion exchange chromatography on DEAE-Sepharose was able to separate the polysaccharides from the enzyme, which was recovered in the elution step, maintaining its enzymatic activity. The batch adsorption of bromelain was evaluated in terms of total protein and enzymatic activity using Langmuir and Langmuir-Freundlich models. Results showed that the process could be suitable for the recovery and purification of the enzyme, maintaining its specific activity.971979Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq