Sequential encapsulation of enzymes within the P22 virus-like particles indicates maturation of an enzyme is important to obtaining optimal kinetic activity

• In Event: Poster Presentation and Judging for: Biomedical; Cell & Molecular Biology; Chemistry & Biochemistry; Marine Science; Systematics & Evolutionary Biology.
  In Poster Session: Chemistry & Biochemistry Poster Session

Abstract

Encapsulation of enzymes into protein cage structures holds promise for better understanding of enzyme function in cellular environments by mimicking crowding conditions and for developing new catalytic nanomaterials. While encapsulation inside protein cages has been shown to modulate enzymatic activity, it is unclear whether these changes in kinetic activity are due to crowding effects or the result of modified folding and maturation of the enzyme. The research presented here evaluates the enzymatic activity of formate dehydrogenase, from Candida boidinii, and alcohol dehydrogenase D, from Pyrococcus furiosus, when encapsulated inside protein cage virus-like particles (VLPs) derived from bacteriophage P22. Results from a rapid in vivo genetically programmed encapsulation strategy showed greatly reduced kinetic activity (kcat) upon protein expression. To examine whether these kinetic changes were the result of crowding effects or interference of the folding and/or maturation of the enzyme, a modified in vivo encapsulation strategy was implemented that enabled the enzyme to first be expressed, allowing folding and maturation, then encapsulated within the P22 VLP. Results suggest some enzymes may require maturation to their active conformation before encapsulation inside VLPs to obtain optimally active enzymes and that kinetic activity changes are not entirely the result of cellular crowding effects as previously assumed. The strategy and results reported highlight important considerations for the encapsulation of enzymes inside confined reaction environments towards the development of active functional nano-reactors.

Authors

• Christy Hjorth, The University of Texas at Tyler

• Dustin Patterson, The University of Texas at Tyler