PARTIAL-PURIFICATION OF GLUTARYL-7-ACA ACYLASE FROM CRUDE CELLULAR LYSATE BY REVERSE MICELLES

Glutaryl-7-ACA acylase was partially purified from the cellular lysate of Pseudomonas sp. NCIMB 40409 by means of reverse micelles-water-two -phases extractions. The tetrameric enzyme can be solubilized inside t he reverse micelles formed by anionic (Aerosol OT, AOT) and cationic ( tetradecyltrimethylammoniumbromide, TDAB) surfactants with retention o f the enzymatic activity. With TDAB reverse micelles system, the acyla se was partially extracted from the aqueous phase and, after backward transfer into a second water phase, a twofold purification factor was achieved. On the other hand, with the AOT micellar system, in conditio ns were most of the proteins but acylase, were extracted by the organi c micellar solution, a sixfold increase of the specific activity of th e acylase remaining in the aqueous phase was obtained.