Covalent immobilization of lipase from Candida rugosa on Eupergit.docVIP

UDC 577.15:663.18 BIBLID: 1450–7188 (2005) 36, 179-186 APTEFF, 36, 1-266 (2005) Original scienti?c paper COVALENT IMMOBILIZATION OF LIPASE FROM Candida rugosa ON EUPERGIT ? Dejan I. Bezbradica, Jasmina J. ?orovi?, Radivoje M. Prodanovi?, Nenad B. Milosavi? and Zorica D. Kne?evi? An approach is presented for the stable covalent immobilization of lipase from Can- dida rugosa on Eupergit with a high retention of hydrolytic activity. It comprises cova- ? lent bonding via lipase carbohydrate moiety previously modi?ed by periodate oxidation, allowing a reduction in the involvement of the enzyme functional groups that are proba- bly important in the catalytic mechanism. The hydrolytic activities of the lipase immobi- lized on Eupergit by two conventional methods (via oxirane group and via glutaralde- ? hyde) and with periodate method were compared. Results of lipase assays suggest that periodate method is superior for lipase immobilization on Eupergit among methods ap- ? plied in this study with respect to both, yield of immobilization and hydrolytic activity of the immobilized enzyme. KEY WORDS: Eupergit ? ; lipase; Candida rugosa; immobilization INTRODUCTION Lipase (E.C. 3.1.1.3.) gained enormous attention in scienti?c community during the last two decades due to a variety of commercial products that can be obtained in lipase- catalyzed reactions (1-3). The most important products are diglycerides, monoglycerides, fatty acids and glycerol (products of fat and oil hydrolysis), ?avor esters and sugar esters (products of esteri?cation) and puri?ed enantiomers with improved physiological action. A signi?cant advance in the application of lipases was the discovery of enhanced lipase activity and thermal stability in reactions carried out in organic solvents, especially those with low polarity (1). Another improvement occurred due to the great progress in the ?eld of development new techniques and novel carrier materials for enzyme immobilization Dejan I.