IMPROVED PURIFICATION PROTOCOL OF A FUSARIUM-SOLANI PISI RECOMBINANT CUTINASE BY PHASE PARTITIONING IN AQUEOUS 2-PHASE SYSTEMS OF POLYETHYLENE-GLYCOL AND PHOSPHATE

Recombinant Fusarium solani pisi cutinase is generally purified by a m ultistep process that includes (a) osmotic shock, (b) acid precipitati on, and chromatography. Steps a and b provide a combined yield of only 77% with a purification factor of 4.1. We report a single-step proces s using an aqueous two-phase partitioning system of 25% (w/w) polyethy lene glycol 1000 and 10% (w/w) sodium phosphate, pH 4.8, to obtain 91% yield with a purification factor of 4.1. This protocol is completed w ithin 20 min, is a cheap alternative to steps a and b, and is easy to scale up.