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Molecular Packaging of Biocatalysts Using a Robust Protein Cage
preprintsubmitted on 02.04.2020, 10:13 and posted on 03.04.2020, 11:06 by Nikola Lončar, Henriette J. Rozeboom, Linda E. Franken, Marc C. A. Stuart, Marco Fraaije
In this paper, we report on the discovery of a novel, robust protein cage (encapsulin) that we could use for packaging various biocatalysts. We have elucidated the structure of the stable encapsulin by electron microscopy and X-ray diffraction. Furthermore, we developed an effective expression system for the encapsulin and a facile protocol for preparing encapsulated enzymes. By packaging and testing various enzymes (varying in size, oligomeric structure, and cofactor type) we demonstrate that, through encapsulation, the enzymes become significantly more stable. We also provide evidence that the pores of the encapsulin, through a size-exclusion effect, can modulate the substrate acceptance profile of an encapsulated enzyme.