INVESTIGADORES
NERLI Bibiana Beatriz
artículos
Título:
Optimization of pancreatic trypsin extraction in PEG/citrate aqueous two-phase systems
Autor/es:
PÉREZ, ROCÍO; LOUREIRO, DANA; NERLI, BIBIANA; TUBÍO, GISELA
Revista:
PROTEIN EXPRESSION AND PURIFICATION
Editorial:
ACADEMIC PRESS INC ELSEVIER SCIENCE
Referencias:
Lugar: Amsterdam; Año: 2015 vol. 106 p. 66 - 71
ISSN:
1046-5928
Resumen:
Enzyme extraction using aqueous two-phase systems (ATPS) has been increasingly used as a primary recovery technique which integrates the clarification, concentration and partial purification of important biomolecules from their natural source in a single step. The goal of this work was to optimize the extraction of trypsin from pancreas homogenate with polyethylene glycol and sodium citrate (PEG/NaCit) ATPS by using the tools of experimental design. The variables NaCl concentration ? added inert salt , the top/bottom phase volume ratio ? Vr ? and the biomass loaded into the system ? in percentage ? were selected as the main factors in the trypsin extraction. The yield (%) and the purification factor of trypsin were considered the responses to be optimized. The central composite design and the response surface analysis proved to be suitable tools for a quick and efficient study. As a result, the optimal extraction conditions in PEG3350/NaCit system were 3.34% wt/wt for NaCl concentration, a biomass load which represented 9.30% wt/wt of the total ATPS mass and 6.37 top/bottom volume ratio giving a purification factor of 2.55 and a yield of 99.7% in top phase.