Strategies of covalent immobilization of a recombinant Candida antarctica lipase B on pore-expanded SBA-15 and its application in the kinetic resolution of (R,S)-Phenylethyl acetate

• Published in: Journal of molecular catalysis. B, Enzymatic Enzymatic, 2016-11, Vol.133, p.246-258
• Main Authors: Rios, Nathalia S., Pinheiro, Maisa P., dos Santos, José Cleiton S., de S. Fonseca, Thiago, Lima, Lara D., de Mattos, Marcos C., Freire, Denise M.G., da Silva, Ivanildo J., Rodríguez-Aguado, Elena, Gonçalves, Luciana R.B.
• Format: Article
• Language: English
• Subjects: Candida antarctica lipase B; Chemical modification; Enzyme immobilization; Modified SBA-15
• ISSN: 1381-1177; 1873-3158
• DOI: 10.1016/j.molcatb.2016.08.009

[Display omitted] •LIPB was successfully immobilized on modified SBA-15 supports with DVS and GA.•Biocatalysts produced are more stable than Glyoxyl-agarose-LIPB preparations.•All the biocatalysts are able to catalyze the resolution of rac-Phenylethyl acetate.•Multipoint covalent preparation increased enzyme rigidity, improving its stability.•Biocatalyst 2.2 is efficient in resolution of rac-Phenylethyl acetate for 5 cycles. A recombinant Candida antarctica lipase B expressed in Pichia pastoris (LIPB) was immobilized on pore-expanded SBA-15 previously modified 3-amino-propyltriethoxysilane (APTES) and activated with two bifunctional reagents, glutaraldehyde (GA) or divinylsulfone (DVS), producing the biocatalysts: SBA-15-APTES-GA-LIPB and SBA-15-APTES-DVS-LIPB, respectively. After LIPB immobilization, both preparations were then modified with glutaraldehyde, producing the biocatalysts: SBA-15-APTES-GA-LIPB-GA, SBA-15-APTES-DVS-LIPB-DVS. Alternatively, LIPB was immobilized on SBA-15-APTES-DVS at pH 10.2 and the biocatalyst was named SBA-15-APTES-DVS-LIPB-pH10. The different biocatalysts were assayed to check the effect of the immobilization strategies on the stability and in the substrate specificity during the kinetic resolution of (R,S)-Phenylethyl acetate. The thermal stability of some new preparations were higher than LIPB adsorbed on SBA-15 (SBA-15-LIPB) and LIPB immobilized on Glyoxyl-agarose. High conversions in the enzymatic kinetic resolution were obtained (43–50%) for all biocatalysts studied. Regarding activity and stability, the SBA-15-APTES-DVS-LIPB-pH10 was the most successful strategy, since, in first cycle, the maximum conversion was obtained (50%), and the biocatalyst remained active and enantioselective even after five successive cycles.