(620bv) Enzyme Encapsulation Using Cell Free Protein Synthesis

Enzyme Encapsulation using Cell Free Protein Synthesis

Seung-Ook Yang, Bradley C. Bundy

Abstract

Nature has evolved to bear a micro-compartmentalization strategy for a number of various functions, such as protection and delivery of cargo molecules, regulation of chemical reaction, assisting proper folding of proteins, etc. These characteristics interest many biotechnologists to utilize viral and non-viral capsids to mimic nature for many applications. To date, a small number of viral capsids have been used to encompass enzymes to build nanoreactors. For example, Bacteriophage Q-beta’s coat protein  self-assembles into a 180-membered virus capsid. Recent studies show this capsid has an ability to encapsulate enzymes when co-produced.  Substrates and product diffuse through pores in the capsid, while the encapsulated enzyme is protected and stabilized from biologically harsh conditions. Here we demonstrate the utility of a cell free protein synthesis (CFPS) environment for improved control over the encapsulation of enzymes. CFPS is in vitro protein synthesis where reagents for protein syntheses can be directly controlled, cytotoxicity is no longer an issue, and expression/folding rates can be tuned and engineered.  Here we report the encapsulation of Lipase B from Candida antarctica in the Q-beta virus capsid using CFPS.