The virus like particle (VLP) derived from bacteriophage P22 presents a unique platform for constructing catalytically functional nanomaterials by directed encapsulation of enzymes into its interior volume of the capsid. Enzyme encapsulation has been engineered to be genetically programmed allowing �one pot� biosynthesis and directed self-assembly of desired enzymes within the roughly 60 nm diameter P22 capsid. The resulting nano-reactors comprise multiple copies of the cargo enzymes, densely packaged within the capsid at local concentrations that mimic predicted high intracellular macromolecule concentrations. Using enzymes, derived from the hyperthermophile Pyrococcus furiosis, we have encapsulated a three enzyme pathway, for carbohydrate metabolism, within the P22 capsid through a process of directed self-assembly. The resulting nano-reactors provide a proof-of-principle for the bioengineering of robust and complex catalytic nanomaterials. The P22 capsid additionally undergoes a series of thermally induced morphological transitions (at 65 and 75�C) that alter the overall volume and porosity of the VLP structure. The system thus provides a platform with which to interrogate the effects of protein crowding on enzyme activity, the importance of adjacencies and diffusion of intermediate species between partner enzymes in model metabolic pathways, and the effects of the capsid as a potential barrier limiting substrate access between the bulk solution and the encapsulated enzymes. Aspects of the structure and kinetic behavior of these systems have been elucidated and a diffusion based model for the enzyme cascade reactions has been developed.The findings reported here clearly illustrate the potential for synthetic approaches to create nano-reactors, by design, using the power of highly evolved and stable enzymes for chemical transformations.

Protein cage nanoreactors: Synthetic enzyme cascades derived from the hyperthermophile <i>Pyrococcus furiosis</i>

248th National Meeting (2014)

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Protein cage nanoreactors: Synthetic enzyme cascades derived from the hyperthermophile Pyrococcus furiosis