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Tuning the catalytic properties of P22 nanoreactors through compositional control

Abstract

Enzymes are biomacromolecular protein catalysts that are widely used in the plethora of industrial-scale applications due to their high selectivity, efficiency and ability to work under mild conditions. Many industrial processes require the immobilization of enzymes to enhance their performance and stability. Encapsulation of enzymes in protein cages provides an excellent immobilization platform to create nanoreactors with enhanced enzymatic stability and desired catalytic activities. Here we show that the catalytic activity of nanoreactors, derived from the bacteriophage P22 viral capsids, can be finely-tuned by controlling the packaging stoichiometry and packing density of encapsulated enzymes. The packaging stoichiometry of the enzyme alcohol dehydrogenase (AdhD) was controlled by co-encapsulating it with wild-type scaffold protein (wtSP) at different stoichiometric ratios using an in vitro assembly approach and the packing density was controlled by selectively removing wtSP from the assembled nanoreactors. An inverse relationship was observed between the catalytic activity (kcat) of AdhD enzyme and the concentration of co-encapsulated wtSP. Selective removal of the wtSP resulted in the similar activity of AdhD in all nanoreactors despite the difference in the volume occupied by enzymes inside nanoreactors, indicating that the AdhD enzymes do not experience self-crowding even under a high molarity of confinement (Mconf) conditions. The approach demonstrated here not only allowed us to tailor the activity of encapsulated AdhD catalysts but also the overall functional output of nanoreactors (enzyme-VLP complex). The approach also allowed us to differentiate the effects of crowding and confinement on the functional properties of enzymes encapsulated in an enclosed system, which could pave the way for designing more efficient nanoreactors.

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Publication details

The article was received on 27 Sep 2019, accepted on 02 Dec 2019 and first published on 03 Dec 2019


Article type: Paper
DOI: 10.1039/C9NR08348K
Nanoscale, 2019, Accepted Manuscript

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    Tuning the catalytic properties of P22 nanoreactors through compositional control

    J. Sharma and T. Douglas, Nanoscale, 2019, Accepted Manuscript , DOI: 10.1039/C9NR08348K

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