Modification of CCMV nanocages for enzyme encapsulation

• Main Authors: Schoonen, Lise, van Hest, Jan C.M.
• Other Authors: Udit, A.K.
• Format: Book Part
• Language: English
• Published: Springer 2018
• Subjects: Candida antarctica lipase B; Cowpea chlorotic mottle virus; Esterase activity; Self-assembly; Sortagging; Sortase A; Viral capsid; Gene Expression; Models; Molecular; Aminoacyltransferases/chemistry; Virus Assembly; Bromovirus; Capsid Proteins/chemistry; Nanocapsules/chemistry; Chromatography; Affinity; Enzymes; Immobilized/chemistry; Capsid/chemistry; Mass Spectrometry; Protein Conformation; Bacterial Proteins/chemistry; Cysteine Endopeptidases/chemistry; Recombinant Fusion Proteins/chemistry; Hydrogen-Ion Concentration; Antarc*; Antarctica
• Online Access: https://research.tue.nl/en/publications/a8a802fb-e2c4-4247-80ec-6599bf8a36e3; https://doi.org/10.1007/978-1-4939-7893-9_6; http://www.scopus.com/inward/record.url?scp=85048174233&partnerID=8YFLogxK

In cellular systems, compartmentalization plays an important role in the protection and regulation of enzymes. Controlled encapsulation of enzymes in nanocompartments is crucial in understanding biocatalytic processes in the cellular environment. We have recently described an enzymatic method to covalently attach enzymes, equipped with a small recognition peptide, to the interior of viral capsids. Viral capsids are especially interesting in this respect, as they form very well-defined nanoparticles with a uniform size and shape. Here, we describe the relevant experimental procedures to encapsulate a model enzyme into the interior of a viral capsid, purify the resulting viral capsids, and measure the catalytic activity of the encapsulated enzymes.