Magnetic biocatalytic nanoreactors based on graphene oxide with graded reduction degrees for the enzymatic synthesis of phytosterol esters

Abstract

As we know, the carriers of enzymatic nanoreactors with different amphipathy have significant influence on the interfacial enzyme activities and its performance. In this study, the magnetic graphene oxide (GO) enzyme carriers were constructed by assembling graphene oxide with different reduction degrees (rGO(x)) and polyethyleneimine (PEI) modified nano-flowery Fe3O4 into composite materials through electrostatic interactions, and x here represents the reduction reaction hours of GO. The SEM, TEM, contact angles and BET tests qualified the ideal enzyme carrier, PEI-Fe3O4-rGO(12), by adsorption-crosslinking method, the Candida rugosa lipase (CRL) was immobilized to obtain the lipase nanoreactor, PEI-Fe3O4-rGO(12)@CRL. The nano-sheet and nano-flower structures provided a larger specific surface area and suitable pore size, and the CRL had higher protein load on the carrier material (163.11 mg/g). In addition, using hydrolyzed p-nitrophenol palmitate as the model, the PEI-Fe3O4-rGO(12)@CRL can still show 76.5 % residual activity after 8 repeated uses. At the same time, the immobilized lipase showed good catalytic performance in the biosynthesis of phytosterol esters. The optimal esterification conditions were determined by single factor experiment (isooctane, sterol concentration 150 μmol/mL, alkyd molar ratio 1:2, enzyme dosage 120 mg, 26 h, 55 °C, the esterification rate was 76.24 %). Therefore, the optimal biocatalyst PEI-Fe3O4-rGO(12)@CRL may has good performance in the biosynthesis of other novel foods.