HYDROPHOBIC INTERACTION CHROMATOGRAPHY OF CHROMOBACTERIUM-VISCOSUM LIPASE

The influence of mobile phase composition on the chromatographic behav iour of Chromobacterium viscosum lipase was studied by using an epoxy- activated spacer arm as a ligand in hydrophobic interaction chromatogr aphy. The retention of lipase depends on the salt used and increased w ith ionic strength, indicating that the interaction of lipase with the stationary phase is of a hydrophobic nature. Using 20% (w/v) ammonium sulphate in the eluent a total retention of lipase on the column was obtained and by washing with 10 mM phosphate buffer a recovery of 79% protein and 89% lipolytic activity were achieved.