Toggle Main Menu Toggle Search

Open Access padlockePrints

Extracellular Self-Assembly of Functional and Tunable Protein Conjugates from Bacillus subtilis

Lookup NU author(s): Professor Colin Harwood

Downloads


Licence

This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

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.


Publication metadata

Author(s): Gilbert C, Howarth M, Harwood CR, Ellis T

Publication type: Article

Publication status: Published

Journal: ACS Synthetic Biology

Year: 2017

Volume: 6

Pages: 957-967

Online publication date: 23/02/2017

Acceptance date: 10/02/2017

Date deposited: 08/12/2017

ISSN (electronic): 2161-5063

Publisher: American Chemical Society

URL: https://doi.org/10.1021/acssynbio.6b00292

DOI: 10.1021/acssynbio.6b00292


Altmetrics

Altmetrics provided by Altmetric


Actions

Find at Newcastle University icon    Link to this publication


Share