Engineering Escherichia coli for enhanced sensitivity to the autoinducer-2 quorum sensing signal.

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TitleEngineering Escherichia coli for enhanced sensitivity to the autoinducer-2 quorum sensing signal.
Publication TypeJournal Article
Year of Publication2019
AuthorsStephens, K, Zargar, A, Emamian, M, Abutaleb, N, Choi, E, Quan, DN, Payne, GF, Bentley, WE
JournalBiotechnol Prog
Paginatione2881
Date Published2019 Jul 15
ISSN1520-6033
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.

DOI10.1002/btpr.2881
Alternate JournalBiotechnol. Prog.
PubMed ID31306566
Grant ListHDTRA1-13-0037 / / Defense Threat Reduction Agency /
R21EB024102 / EB / NIBIB NIH HHS / United States
CBET 1805274 / / National Science Foundation /
DMREF 1435957 / / National Science Foundation /
ECCS 1807604 / / National Science Foundation /