LIPASE OF PSEUDOMONAS-CEPACIA FOR BIOTECHNOLOGICAL PURPOSES - PURIFICATION, CRYSTALLIZATION AND CHARACTERIZATION

Commercial lipase (triacylglycerol lipase, EC 3.1.1.3) of Pseudomonas cepacia (Amano) has been purified to homogeneity by a single chromatog raphy on phenyl Sepharose. The eluted lipase crystallized spontaneousl y at 4 degrees C in the eluent, containing 58-69% 2-propanol. The yiel d of the lipase was 87-100% and the specific activity during the hydro lysis of triolein 5800 U/mg protein. This protein has a molecular weig ht of 34.1 kDa as analyzed by sodium dodecyl sulfate-polyacrylamide ge l electrophoresis (SDS-PAGE). Its purity was determined by SDS-PAGE an d capillary zone electrophoresis to be greater than or equal to 99%. I mmobilization on Sepharose increased its stability in organic solvents . This lipase of P. cepacia differs from that of other Pseudomonas str ains in respect to substrate specificity and during crystallization. I t exhibits a high stability in organic solvents and supercritical carb on dioxide.