1,720,980 research outputs found
A friendly method for Raphanus sativus L (wild radish) peroxidase purification by polyelectrolyte precipitation
Full text linkThe separation of radish peroxidase from a fresh Raphanus sativus L extract was carried out using precipitation with two commercially available negatively charged synthetic polyelectrolytes: Eudragit® L 100 and Eudragit® S 100. The enzyme was precipitated by polyelectrolyte addition at pH 4.00. The non-soluble complex formed was separated by simple centrifugation and re-dissolved by a pH change. The recovery of radish peroxidase biological activity was 50% of the initial activity in the homogenate for EuL and 45% for EuS, with 1.5-fold increase in its specific activity. The total Eudragit® concentration to precipitate the enzyme was very low: about 2 × 10 -3% w/v. The volume of the final product decreased to 10% of the feedstock, concentrating the sample up to 10 times.Fil: Woitovich Valetti, Nadia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química y Física. Área Fisicoquímica; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química y Física. Área Fisicoquímica; Argentin
Characterization of chymotrypsin-ι-carrageenan complex in aqueous solution: A solubility and thermodynamical stability study
Full text linkThe aim of this study is to report the results of research work on the molecular mechanism of complex formation between chymotrypsin and a negatively charged natural strong polyelectrolyte, ι-carrageenan, using spectroscopy techniques. The carrageenan-chymotrypsin complex showed a maximal non-solubility at pH around 4.50 with a stoichiometric ratio between 8 and 33. g of chymotrypsin per g of carrageenan. These values were depended on the enzyme concentration, pH and ionic strength medium. The insoluble complex was redissolved by modifying the pH and by a NaCl concentration around 0.2. M in agreement with a coulombic mechanism of complex formation. The non-soluble complex formation showed biphasic kinetics. A fast step was carried out around 10. s and a coulombic mechanism takes place, and a slower step of around 120. s, where participate only Van der Waals forces. The enzymatic activity of chymotrypsin was maintained even in the presence of carrageenan (0.005%, w/v).Fil: Woitovich Valetti, Nadia. Universidad Nacional de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Boeris, Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario; ArgentinaFil: Picó, Guillermo Alfredo. Universidad Nacional de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentin
Adsorption isotherms, kinetics and thermodynamic studies towards understanding the interaction between cross-linked alginate-guar gum matrix and chymotrypsin
No full textThe adsorption kinetics of chymotrypsin, a pancreatic serine protease, onto an alginate-gum guar matrix cross-linked with epichlorohydrin has been performed using a batch-adsorption technique. The effect of various experimental parameters such as pH, salt presence, contact time and temperature were investigated. The pseudo-first-order and pseudo-second-order kinetic models were used to describe the kinetic data which shows that the adsorption of the enzyme followed the pseudo-second-order rate expression. The Langmuir, Freundlich and Hill adsorption isotherm models were applied to describe the equilibrium isotherms, and the isotherm constants were determined. It was found that Hill model was more suitable for our data because the isotherm data showed a sigmoidal behavior with the free enzyme concentration increasing in equilibrium. At 8 °C and at pH 5.0, 1 g hydrate matrix adsorbed about 7 mg of chymotrypsin. In the desorption process 80% of the biological activity of chymotrypsin was recovered under the condition of 50 mM phosphate buffer, pH 7.00-500 mM NaCl. When successive cycles of adsorption/washing/desorption were performed, it was observed that the matrix remained functional until the fourth cycle of repeated batch enzyme adsorption. These results are important in terms of diminishing of cost and waste generation.Fil: Woitovich Valetti, Nadia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin
Prediction of breakthrough curves in packed-bed column as tool for lysozyme isolation using a green bed
No full textThe adsorption of lysozyme onto alginate–guar gum bed cross-linked with epichlorohydrin has been studied in packed-bed column. Adsorption performance was evaluated with different bed heights, flow rates and initial protein concentrations. The Thomas and bed depth service time mathematical models fitted well the breakthrough curves experimental data with high correlation coefficients. Not modification of the working yield of the bed was observed in progressive ten cycles of reused. Lysozyme recovery of 75% was achieved from white egg with a purification factor around 15 under the condition of bed height 9 cm, flow rate 0.4 mL/min and total protein inlet 10 mg/mL, while the dynamic bed capacity under this experimental condition was 10.87 mg/g hydrated bed.Fil: Brassesco, Maria Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Woitovich Valetti, Nadia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin
Molecular mechanism of lysozyme adsorption onto chemically modified alginate guar gum matrix
Full text linkThe equilibrium isotherms and adsorption kinetics of lysozyme (LZ) on epichlorohydrin (Epi) cross-linked alginate-guar gum (Alg-GG) matrix were studied. Adsorption kinetics followed a pseudo-first-order model while the equilibrium isotherm could be represented by the Freundlich equation. The maximal amount of LZ adsorbed onto this matrix was around 2.4 mg per g of hydrated matrix at pH 7.00. The adsorption mechanism was associated to a simple diffusion process with a weak columbic interaction between LZ and the matrix. The presence of NaCl 0.3 M induced a total displacement of the LZ from the matrix. Under this condition, the percentage of desorbed protein was 95%. Successive cycles of adsorption-washing-elution were performed and the results showed the reversibility of the process and the usefulness of the method for enzyme purification and separation. A last successful step was carried out for the purification of LZ from egg white as natural source. The model proved to be useful applied as a platform design in the isolation and purification of proteins.Fil: Brassesco, Maria Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Woitovich Valetti, Nadia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin
A new platform for chymotrypsin isolation from fresh bovine pancreas using an environmental friendly polyelectrolyte: Alginate
Full text linkThe separation of chymotrypsin from a crude filtrate of fresh bovinepancreas homogenate was carried out using precipitation with acommercially available negatively charged natural weak polyelectrolyte:sodium alginate. The zymogen form of the enzyme was activated by theaddition of trypsin at pH 8.2 in the absence of Ca++, then, the enzyme wasprecipitated by sodium alginate addition at pH 5.00. The non-solublecomplex was separated by simple centrifugation and re-dissolved by apH change to 8.20. The recovery of chymotrypsin biological activity was36.1 % of the initial activity in the pancreas homogenate with a 3.2 foldincrease in its specific activity. The volume of the final product decreasedto 6.25 % of the initial feedstock, concentrating the sample up to 16times.Fil: Rausch, Caren. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Tecnología; ArgentinaFil: Woitovich Valetti, Nadia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Tecnología; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Tecnología; Argentin
Use of renewable feedstocks: The recovery of high value molecules from waste of renewable feedstocks: soybean hull
No full textIn the last 20 years intensive crop farming has been developed in a significantly manner in Latin America, whereas the cultivation of one or more type of crops have been increasing in each country. Within these crops, the most common ones are: soybeans, rice, corn, wheat, coffee, sugarcane, etc. The industrialization of the agriculture biomass produces millions of tons of waste that are discarded into the environment. Soybean is the main crop in Argentina and Brazil, producing about 140 million of Tons /year. Its industrialization produces soybean hull which is a waste only used as additive for cattle feed. This chapter provides an updated review about the application and reconversion of this waste, referencing to the high value molecules which it contains, due to their application in different areas of biotechnology: residual water treatment, dietary fiber, ethanol production, obtainment of: monosaccharides, polysaccharides, cellulose and proteins with important biological activity such as peroxidase, serin proteases and proteins with antitumor activity.Fil: Camiscia, Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Woitovich Valetti, Nadia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin
Precipitation of chymotrypsin from fresh bovine pancreas using i-carrageenan
No full textThe separation of chymotrypsin from a crude filtrate of fresh bovine pancreas homogenate was carried out using precipitation with a commercially available negatively charged natural strong polyelectrolyte: -carrageenan. The zymogen form of the enzyme was activated by addition of trypsin (0.0001%, w/w), then, the enzyme was precipitated by polyelectrolyte addition at pH 4.50. The non-soluble complex was separated by simple centrifugation and re-dissolved by a pH change to 8.20. The recovery of chymotrypsin biological activity was 60% of the initial activity in the homogenate with 3-fold increase in its specific activity. The volume of the final product decreased to 10% of the feedstock, concentrating the sample up to 10 times.Fil: Woitovich Valetti, Nadia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química y Física. Área Fisicoquímica; ArgentinaFil: Lombardi, Julia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química y Física. Área Fisicoquímica; ArgentinaFil: Boeris, Valeria. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química y Física. Área Fisicoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Departamento de Química y Física. Área Fisicoquímica; Argentin
Obtainment of a highly concentrated pancreatic serine proteases extract from bovine pancreas by precipitation with polyacrylate
Full text linkSerine proteases have wide application in leather, food, meat and soap powder industries, among others. There are a lot of methodologies to obtain them in large quantities but most of them are expensive or contaminating. We then propose an economical and environmentally friendly method in which proteases are separated from a crude fresh bovine pancreas homogenate using precipitation with polyacrylate, a commercially available negatively charged and weak polyelectrolyte. The zymogens of the serine proteases were activated prior to precipitation by the addition of trypsin. The proteases were precipitated by adding polyacrylate at pH 4.50 to the pancreas homogenate. Under these conditions, serine proteases are positively charged and form non-soluble complexes with the negatively charged polyacrylate. The purity of proteases was increased 5-fold with a recovery of 33% under the best conditions tested. The volume of the final product was decreased to 5% of the feedstock, in order to concentrate the sample up to 20 times. The proposed method removed up to 96% of the contaminant proteins.Fil: Lombardi, Julia. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Cientifico Tecnológico Rosario; ArgentinaFil: Woitovich Valetti, Nadia. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Cientifico Tecnológico Rosario; ArgentinaFil: Picó, Guillermo Alfredo. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Cientifico Tecnológico Rosario; ArgentinaFil: Boeris, Valeria. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Cientifico Tecnológico Rosario; Argentin
Extraction and purification of peroxidase and trypsin inhibitor from soybean hulls: A strategy to revalue a waste as a source of different types of molecules of biotechnological interest
No full textIn this work, the extraction and purification of proteins present in soybean hull has been carried out, using simple methods and with the potential to be scaled. In the first stage, using statistical tools, the optimization of peroxidase extraction and various protease inhibitor factors was achieved through a leaching process using 50 mM phosphate buffer pH 6.00. The optimal conditions to achieve the best extraction of the proteins of interest were the following: contact time (360 min), mass/volume ratio (0.066) and particle size (40 mesh). The extract obtained was characterized by an electrophoresis gel, mass spectrometry and by measuring enzymatic activity, which, allowed to confirm the presence of the proteins of interest. Finally, the purification of peroxidase andprotease inhibitors were optimized by adsorption onto aminated cellulose bed obtained from the same soybean hull. Under the optimal conditions, the peroxidase present in the extract was purified 8.1 times with a yield of 66.6%. While protease inhibitors were purified 2.5 times with a yield of 26.1%.Fil: Camiscia, Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Silva, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Woitovich Valetti, Nadia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin
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