Engineering Escherichia coli for enhanced sensitivity to the autoinducer‐2 quorum sensing signal
Abstract
The autoinducer‐2 (AI‐2) quorum sensing system is involved in a range of population‐based bacterial behaviors and has been engineered for cell–cell communication in synthetic biology systems. Investigation into the cellular mechanisms of AI‐2 processing has determined that overexpression of uptake genes increases AI‐2 uptake rate, and genomic deletions of degradation genes lowers the AI‐2 level required for activation of reporter genes. Here, we combine these two strategies to engineer an Escherichia coli strain with enhanced ability to detect and respond to AI‐2. In an E. coli strain that does not produce AI‐2, we monitored AI‐2 uptake and reporter protein expression in a strain that overproduced the AI‐2 uptake or phosphorylation units LsrACDB or LsrK, a strain with the deletion of AI‐2 degradation units LsrF and LsrG, and an “enhanced” strain with both overproduction of AI‐2 uptake and deletion of AI‐2 degradation elements. By adding up to 40 μM AI‐2 to growing cell cultures, we determine that this “enhanced” AI‐2 sensitive strain both uptakes AI‐2 more rapidly and responds with increased reporter protein expression than the others. This work expands the toolbox for manipulating AI‐2 quorum sensing processes both in native environments and for synthetic biology applications.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Aug 10, 2019
- Source ID
- 10.1002/btpr.2881
Entities
People
- Amin Zargar
- David N. Quan
- Erica Choi
- Gregory F Payne
- Kristina Stephens
- Milad Emamian
- Nadia Abutaleb
- William E. Bentley
Organizations
- Defense Threat Reduction Agency
- National Institute of Biomedical Imaging and Bioengineering
- National Science Foundation
- University of Maryland