Modification of PEGylated enzyme with glutaraldehyde can enhance stability while avoiding intermolecular crosslinking

Abstract

We demonstrate an enzyme stabilization approach whereby a model enzyme is PEGylated, followed by controlled chemical modification with glutaraldehyde. Using this stabilization strategy, size increases and aggregation due to intermolecular crosslinking are avoided. Immediately following synthesis, the PEGylated enzyme with and without glutaraldehyde modification possessed specific activities of 372.9 ± 20.68 U mg⁻¹ and 373.9 ± 15.14 U mg⁻¹, respectively (vs. 317.7 ± 19.31 U mg⁻¹ for the native enzyme). The glutaraldehyde-modified PEGylated enzyme retains 73% original activity after 4 weeks at 37 °C (vs. 8.2% retention for control).