Ternary biogenic silica/magnetite/graphene oxide composite for the hyperactivation of Candida rugosa lipase in the esterification production of ethyl valerate

Abstract

Oil palm leaves (OPL) silica (SiO₂) can replace the energy-intensive, commercially produced SiO₂. Moreover, the agronomically sourced biogenic SiO₂ is more biocompatible and cost-effective enzyme support, which properties could be improved by the addition of magnetite (Fe₃O₄) and graphene oxide (GO) to yield better ternary support to immobilize enzymes, i.e., Candida rugosa lipase (CRL). This study aimed to optimize the Candida rugosa lipase (CRL immobilization onto the ternary OPL-silica-magnetite (Fe₃O₄)-GO (SiO₂/Fe₃O₄/GO) support, for use as biocatalyst for ethyl valerate (EV) production. Notably, this is the first study detailing the CRL/SiO₂/Fe3O4/GO biocatalyst preparation for rapid and high yield production of ethyl valerate (EV). AFM and FESEM micrographs revealed globules of CRL covalently bound to GL-A-SiO₂/Fe₃O₄/GO; similar to Raman and UV-spectroscopy results. FTIR spectra revealed amide bonds at 3478 cm^-1 and 1640 cm^-1 from covalent interactions between CRL and GL-A-SiO₂/Fe₃O₄/GO. Optimum immobilization conditions were 4% (v/v) glutaraldehyde, 8 mg/mL CRL, at 16 h stirring in 150 mM NaCl at 30 °C, offering 24.78 ± 0.26 mg/g protein (specific activity = 65.24 ± 0.88 U/g). The CRL/SiO₂/Fe₃O₄/GO yielded 77.43 ± 1.04 % of EV compared to free CRL (48.75 ± 0.70 %), verifying the suitability of SiO₂/Fe₃O₄/GO to hyperactivate and stabilize CRL for satisfactory EV production.

Keywords: Biogenic silica; Covalent immobilization; Graphene oxide; Lipase; Magnetite nanoparticle; Ternary composite.