Stabilization of Candida rugosa lipase on nanosized zirconia-based materials

• Published in: Journal of molecular catalysis. B, Enzymatic Vol. 108; pp. 43 - 50
• Main Authors: Guncheva, Maya, Paunova, Krasimira, Dimitrov, Momtchil, Yancheva, Denitsa
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01-10-2014; Elsevier
• Subjects: Bioconversions. Hemisynthesis; Biological and medical sciences; Biotechnology; Candida rugosa lipase; Enantioselectivity; Enzyme stabilization; Fundamental and applied biological sciences. Psychology; Methods. Procedures. Technologies; Nanosized zirconia-based materials; Enzyme stabilization; Enantioselectivity; Candida rugosa lipase; Nanosized zirconia-based materials; Yeast; Enzyme; Stabilization; Materials; Triacylglycerol lipase; Esterases; Carboxylic ester hydrolases; Fungi; Zirconia; Hydrolases; Fungi Imperfecti; Candida rugosa
• ISSN: 1381-1177; 1873-3158
• DOI: 10.1016/j.molcatb.2014.06.012

•Nanosized materials on the basis of zirconia were synthesized and characterized.•Lipase from Candida rugosa (CRL) was immobilized on the novel supports.•Immobilization efficiency depended on structural properties of the materials.•NanoZrO2-A improved thermal- and solvent stability of CRL.•NanoZrO2-A-CRL exhibited good enantioselectivity in acylation of (±)-menthol.
We synthesized and characterized three novel materials on the basis of zirconia. Despite their three-to six fold higher specific surface area, nanoZrO2-CeO2 (150m2/g) and nanoZrO2-B (296m2/g) they proved to be less effective supports for a lipase from Candida rugosa than nanoZrO2-A. For the last, we achieved protein loading of 23mg/g, distributed in a monolayer, which means that 61% of the carrier surface was occupied by the protein. The immobilized on nanoZrO2-A lipase (nanoZrO2-A-CRL) exhibited enhanced thermal and solvent stability in comparison to the native enzyme. NanoZrO2-A-CRL has a half-life at 55°C of 73min, while for the native enzyme it was only 5.3min. 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 nanoZrO2-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.