Protein vesicles incorporating functional, globular proteins have enormous potential in a number of bio-applications such as micro bioreactors and artificial cells. We have previously designed globule-zipper-polypeptide protein complexes (Globule-ZE/ZR-ELP), consisting of a model fluorescent globular protein (mCherry), high affinity leucine zipper pair (ZE/ZR), and thermo-responsive elastin-like polypeptide (ELP). We have shown that the fusion protein complexes undergo a phase transition upon warming, from soluble monomers, through dynamic coacervates, to stable hollow vesicles with different structures and sizes dependent on assembly conditions. In this presentation, I will discuss the use of enzymes as the globular protein domain. We have mapped out the range of surface charge and molecular weight of the globular proteins capable of making protein vesicles. Protein surface charge had little effect on vesicle formation with only small changes in minimum required salt concentration. Not surprisingly, surface charge did affect colloidal stability, with vesicles made from neutral proteins having a slightly higher tendency to aggregate than more highly charged proteins. Molecular weight of the globular enzyme also did not seem to significantly affect vesicle formation in the range of ~20 – 80 kDa, including both monomeric and multimeric enzymes. These results demonstrate that a variety of enzymes could be incorporated into vesicles for retention of structure, recoverability, and product separation benefits. To investigate the potential of enzyme vesicles to provide these features, we utilized a panel of 4 enzymes ranging in size and multimer state, including a chimeric amine dehydrogenase. Enzyme vesicles demonstrated initial catalytic activity similar to soluble enzymes and enzyme stability and vesicle permeability will be shared. Overall, this work provides evidence that the vesicle platform can be used with a wide variety of functional proteins and requires little tuning of the self-assembly conditions (protein concentration, protein stoichiometry, salt concentration, temperature, etc.). Further, it identifies performance benefits of vesicles, which are unique from other enzyme immobilization systems in their reversible assembly/disassembly and compartments with different phases than the bulk solution.

Protein breadth and reactive capability of self-assembled enzyme vesicles

Macromolecular Chemistry: The Second Century

[Author Benefits](https://scimeetings.acs.org/?utm_source=pubs_content_marketing&utm_medium=website&utm_campaign=0320_MCF_NPI_Launch_Spring_Homepage&ref=pubs_content_marketing)

[How to use SciMeetings](https://storage.googleapis.com/dl.morressier.com/acs/SciMeetings-ACSSpring2020-HowToGuide_M3020-Final.pdf)

[Frequently Asked Questions](https://storage.googleapis.com/dl.morressier.com/acs/SciMtng_ACSNMSp2020_FAQ_M3120_Final.pdf)

ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.