Immobilization of enterokinase on magnetic supports for the cleavage of fusion proteins
- Citation:
- Santana, SDF, Pina AS, Roque ACA. 2012. Immobilization of enterokinase on magnetic supports for the cleavage of fusion proteins. Journal of Biotechnology. 161:378–382.
Abstract:
Magnetic nanobiocatalysts for tag cleavage on fusion proteins have been prepared by immobilizing
enterokinase (EK) onto iron oxide magnetic nanoparticles coated with biopolymers. Two different
chemistries have been explored for the covalent coupling of EK, namely carbodiimide (EDC coupling)
and maleimide activation (Sulfo coupling). Upon immobilization, EK initial activity lowered but EDC coupling lead to higher activity retention. Regarding the stability ofthe nanobiocatalysts,thesewere recycled
up to ten times with the greater activity losses observed in the first two cycles. The immobilized EK also
proved to cleave a control fusion protein and to greatly simplify the separation of the enzyme from the
reaction mixture.
