Magnetically-separable and highly-stable enzyme system based on crosslinked enzyme aggregates shipped in magnetite-coated mesoporous silica

Abstract

A magnetically-separable and highly-stable enzyme system was developed by adsorption of enzymes in superparamagnetic hierarchically ordered mesocellular mesoporous silica (M-HMMS) and subsequent enzyme crosslinking. Superparamagnetic nanoparticles were homogeneously incorporated into hierarchically-ordered mesocellular mesoporous silica (HMMS) by the decomposition of a preformed iron propionate complex. The size of the incorporated superparamagnetic nanoparticles was around 5 nm, generating a magnetically separable host with high pore volumes and large pores (M-HMMS). alpha-chymotrypsin (CT) was adsorbed into M-HMMS with high loading (similar to 30 wt%) in less than 30 minutes. Glutaraldehyde (GA) treatment of adsorbed CT resulted in nanometer scale crosslinked enzyme aggregates in M-HMMS (CLEA-M). The activity of these CT aggregates in M-HMMS (CLEA-M-CT) was 34 times than that of simply adsorbed CT in M-HMMS, due to an effective prevention of enzyme leaching during washing via a ship-in-a-bottle approach. CLEA-M-CT maintained the initial activity not only under shaking (250 rpm) for 30 days, but also under recycled uses of 35 times. The same approach was employed for the synthesis of CLEA-M of lipase (CLEA-M-LP), and proven to be effective in improving the loading, activity, and stability of enzyme when compared to those of adsorbed LP in M-HMMS.