title: Efficient extraction of olive pulp and stone proteins by using an enzyme-assisted method.
abstract: An efficient protein extraction protocol for proteins from olive pulp and stone by using enzymes was developed. For this purpose, different parameters that affect the extraction process, such as enzyme type and content, pH, and extraction temperature and time, were tested. The influence of these factors on protein recovery was examined using the standard Bradford assay, while the extracted proteins were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The best extraction conditions were achieved at pH 7.0 and 5% (v/v) Palatase® 20000 L (lipase) for pulp and Lecitase® Ultra (phospholipase) for stone proteins. The optimal extraction temperature and time were 30 and 40 °C for 15 min for pulp and stone tissues, respectively. Under these conditions, several protein extracts coming from olive fruits of different genetic variety were analyzed, their profiles being compared by SDS-PAGE. The developed enzyme-assisted extraction method showed faster extraction, higher recovery, and reduced solvent usage than the nonenzymatic methods previously described in the literature. In the case of stone proteins, different electrophoretic profiles and band intensities were obtained that could be helpful to distinguish samples according to their genetic variety.
authors.affiliation: Dept. of Analytical Chemistry, Univ. of Valencia, C. Doctor Moliner 50, E-46100 Burjassot, Valencia, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Vergara-Barberán
authors.forename: María Jesús
authors.initials: MJ
authors.lastname: Lerma-García
authors.forename: José Manuel
authors.initials: JM
authors.lastname: Herrero-Martínez
authors.forename: Ernesto Francisco
authors.initials: EF
authors.lastname: Simó-Alfonso
mesh_terms: D005591:Chemical Fractionation / Q000379:methods*; D004798:Enzymes / Q000737:chemistry / Q000378:metabolism*; D005638:Fruit / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D031658:Olea / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry*; D013696:Temperature; D013997:Time Factors
chemical_list: D004798:Enzymes; D010940:Plant Proteins
keywords: SDS-PAGE; enzyme-assisted extraction; olive pulp; olive stone; protein profiles