The improvement of protein extraction from olive leaves using an enzyme-assisted protocol has been
investigated. Using a cellulase enzyme (Celluclast 1.5L), different parameters that affect the extraction
process, such as the influence and amount of organic solvent, enzyme amount, pH and extraction temperature
and time, were optimised. The influence of these factors was examined using the standard Bradford
assay and the extracted proteins were characterised by sodium dodecyl sulphate polyacrylamide gel electrophoresis
(SDS–PAGE). The optimum extraction parameters were: 30% acetonitrile, 5% (v/v) Celluclast
1.5L at pH 5.0 and 55 C for 15 min. Under these conditions, several protein extracts from olive leaves of
different genetic variety (with a total protein amount comprised between 1.87 and 6.64 mg g1) were
analysed and compared by SDS–PAGE, showing differences in their electrophoretic protein profiles.
The developed enzyme-assisted extraction method has shown a faster extraction, higher recovery and
reduced solvent usage with respect to the use of the non-enzymatic methods described in literature