METHOD FOR PURIFYING TWO ISOENZYME LIPASES OF $i(CANDIDA RUGOSA)

摘要

Method for the purification of two extracellular lipases present in commercial raw lipase of the yeast Candida rugosa. The method comprises a single stage of hydrophobic chromatography in agarosa matrix. Extremely different elution conditions provide for the complete separation and purification of two enzymes with lipasic activity (lipases A and B) present in the commercial starting extract. To this effect, a concentrated buffer with a pH of 6-8 is used for the elution from the column of a major amount of contaminants present in the extract; thereafter, with the same buffer solution, but more diluted, the lipase B is entrained. Then, with a di or polyalcohol dissolved in the diluted buffer, the lipase A is eluted and by concentration through ultrafiltration and chromatography of melecular exclusion with dextran, the alcohol is removed from said solution. Industrial application to the purification of enzymes.