Continuous hydrolysis of olive oil by lipase immobilized on DEAE-sephadex

Abstract

For the purpose of continuous hydrolysis of fats and lipids immobilized Candida rugosa lipase was used in packed bed reactor system. So as to broaden the substrate range up to solid fat and to enhance the productivity, isooctane was selected as a solvent. By the preliminary repeated batch experiment, DEAE-Sephadex was selected as immobilized matrix (data not shown). The immobilization efficiency was greater than 90\% for the loading capacity up to 20 mg protein/g-wet gel and the maximum loading could be obtained up to 40 mg/g-wet gel with the immobilization efficiency of 70\%. Apparent Km value and optimum pH was 8\% (v/v) olive oil in isooctane and 7. 0, respectively. In order to pack the gel in a column reactor, AOT was used and because of enzyme desorption by AOT, packing method was selected carefully. To increase the operational stability buffer, buffer-solvent ratio and the amount of loaded enzyme were varied. As the results of these experiments triethanol amino buffer was better than phosphate buffer, and less buffer flow rate and more enzyme-loading gel increased the operational stability. Under last condition the half life of immobilized enzyme was increased over 25 times.