Browse by author
Lookup NU author(s): Professor Colin Harwood
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
The ability to stably and specifically conjugate recombinant proteins to one another is a powerful approach for engineering multifunctional enzymes, protein therapeutics and novel biological materials. While many of these applications have been illustrated through in vitro and in vivo intracellular protein conjugation methods, extracellular self-assembly of protein conjugates offers unique advantages: simplifying purification, reducing toxicity and burden, and enabling tenability. Exploiting the recently described SpyTag-SpyCatcher system, we describe here how enzymes and structural proteins can be genetically-encoded to covalently conjugate in culture media following programmable secretion from Bacillus subtilis. Using this novel approach, we demonstrate how self-conjugation of a secreted industrial enzyme, XynA, dramatically increases its resilience to boiling and we show that cellular consortia can be engineered to self-assemble functional protein protein conjugates with tunable composition. This novel genetically-encoded modular system provides a flexible strategy for protein conjugation harnessing the substantial advantages of extracellular self-assembly.
Author(s): Gilbert C, Howarth M, Harwood CR, Ellis T
Publication type: Article
Publication status: Published
Journal: ACS Synthetic Biology
Online publication date: 23/02/2017
Acceptance date: 10/02/2017
Date deposited: 08/12/2017
ISSN (electronic): 2161-5063
Publisher: American Chemical Society
Altmetrics provided by Altmetric