Obtainment of a highly concentrated pancreatic serine proteases extract from bovine pancreas by precipitation with polyacrylate

Authors
Lombardi, Julia; Woitovich Valetti, Nadia; Picó, Guillermo Alfredo; Boeris, Valeria
Publication Year
2013
Language
English
Format
article
Status
Published version
Description
Serine 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; Argentina
Fil: 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; Argentina
Fil: 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; Argentina
Fil: Boeris, Valeria. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Cientifico Tecnológico Rosario; Argentina
Subject
Bovine pancreas proteases
Serine proteases
Trypsin
Chymotrypsin
Polyelectrolyte
Bioprocesamiento Tecnológico, Biocatálisis, Fermentación
Biotecnología Industrial
INGENIERÍAS Y TECNOLOGÍAS
Access level
Restricted access
License
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repository
CONICET Digital (CONICET)
Institution
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identifier
oai:ri.conicet.gov.ar:11336/7471