With the aim of fixing the enzyme to the matrix by multiple covalent linkages, lipase from Candida rugosa (formerly cylindracea) has been insolubilized through its amino groups on Sepharose 6B previously activated with 2,3-epoxy-1-propanol. Two main variables that are known to control the number of bonds formed have been tested: the contact time between enzyme and activated support, and the temperature at which the immobilization reaction is carried out. Studies on activity and stability of the different derivatives prepared showed that higher temperatures and longer contact times lead to insolubilized enzymes that are more resistant to inactivation by temperature and the presence of organic solvents. At 50 degrees C and pH 7.2, the insoluble lipase was found to be 140 times more stable than its soluble counterpart.