Stabilization of Candida rugosa lipase on nanosized zirconia-based materials

作者：Guncheva Maya^*; Paunova Krasimira; Dimitrov Momtchil; Yancheva Denitsa

来源：Journal of Molecular Catalysis B: Enzymatic , 2014, 108: 43-50.

DOI：10.1016/j.molcatb.2014.06.012

• Candida rugosa lipase
• Nanosized zirconia-based materials
• Enzyme stabilization
• Enantioselectivity

We synthesized and characterized three novel materials on the basis of zirconia. Despite their three-to six fold higher specific surface area, nanoZrO(2)-CeO2 (150 m(2)/g) and nanoZrO(2)-B (296 m(2)/g) they proved to be less effective supports for a lipase from Candida rugosa than nanoZrO(2)-A. For the last, we achieved protein loading of 23 mg/g, distributed in a monolayer, which means that 61% of the carrier surface was occupied by the protein. The immobilized on nanoZrO(2)-A lipase (nanoZrO(2)-A-CRL) exhibited enhanced thermal and solvent stability in comparison to the native enzyme. NanoZrO(2)-A-CRL has a half-life at 55 degrees C of 73 min, while for the native enzyme it was only 5.3 mm. The immobilized enzyme preserved 20% of its activity in six consecutive cycles of the reaction hydrolysis of tributyrin. The immobilization influenced the enantioselectivity of CRL Using nanoZrO(2)-A-CRL under optimal reaction condition in the reaction of esterification of lauric acid with (+/-)-menthol, we achieved menthol conversion of 43% (i.e. 82% of (-) methyl laurate), enantiomeric excess of 97% and enantiomeric ratio of 144. The conformational analysis proved that upon immobilization no serious change in the secondary structure of the lipase from Candida rugosa had occurred.

• 出版日期2014-10