Improved immobilization and stabilization of lipase from Rhizomucormiehei on octyl-glyoxyl agarose beads by using CaCl2

FERNANDEZ-LOPEZ, L.; RUEDA, N.; BARTOLOME-CABRERO, R.; RODRÍGUEZ, M. D.; ALBUQUERQUE, T. L.; DOS SANTOS, J. C. S.; BARBOSA, O.; FERNANDEZ-LAFUENTE, R

PROCESS BIOCHEMISTRY - (Print)

ELSEVIER SCI LTD

Lugar: Amsterdam; Año: 2016 vol. 51 p. 48 - 52

1359-5113

A strategy to stabilize octyl (OC)-Rhizomucor miehei lipase (RML) and OC-Candida rugosa lipase (CRL) atpH 10 is necessary to take full advantage of the immobilization of these enzymes on OC-glyoxyl (OCGLX)supports. CaCl2and MnCl2have been reported to stabilize OC-RM and OC-CRL at pH 5.0 and 7.0. Afterscreening different buffers, 5 mM CaCl2was found to be fully soluble in Gly at pH 10. OC-RML was 15folds stabilized at this pH value by CaCl2, while OC-CRL was not. This salt was used in the preparation ofoctyl-glyoxyl (OCGLX)-RML, permitting to maintain almost unaltered the OC-RML activity that was 3 foldhigher than that of the free. This preparation was 30% and 3 folds more stable in thermal or acetonitrileinactivations respectively than the standard OCGLX one.The stabilizing effect of CaCl2and MnCl2on the OCGLX preparations was studied. These salts stabilizedboth OCGLX-RML preparations, although the one prepared using Ca2+during the covalent attachmentwas more stabilized than the standard one by the presence of Ca2+, even 7-8 folds in the presence ofaceonitrile. Thus, this additive permits to recover an OCGLX-RML preparation more stable and activethan the standard protocol.