Characterization of diverse homoserine lactone synthases in Escherichia coli

René Daer, Cassandra M. Barrett, Ernesto Luna Melendez, Jiaqi Wu, Stefan J. Tekel, Jimmy Xu, Brady Dennison, Ryan Muller, Karmella Haynes

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Quorum sensing networks have been identified in over one hundred bacterial species to date. A subset of these networks regulate group behaviors, such as bioluminescence, virulence, and biofilm formation, by sending and receiving small molecules called homoserine lactones (HSLs). Bioengineers have incorporated quorum sensing pathways into genetic circuits to connect logical operations. However, the development of higher-order genetic circuitry is inhibited by crosstalk, in which one quorum sensing network responds to HSLs produced by a different network. Here, we report the construction and characterization of a library of ten synthases including some that are expected to produce HSLs that are incompatible with the Lux pathway, and therefore show no crosstalk. We demonstrated their function in a common lab chassis, Escherichia coli BL21, and in two contexts, liquid and solid agar cultures, using decoupled Sender and Receiver pathways. We observed weak or strong stimulation of a Lux receiver by longer-chain or shorter-chain HSL-generating Senders, respectively. We also considered the under-investigated risk of unintentional release of incompletely deactivated HSLs in biological waste. We found that HSL-enriched media treated with bleach were still bioactive, while autoclaving deactivates LuxR induction. This work represents the most extensive comparison of quorum signaling synthases to date and greatly expands the bacterial signaling toolkit while recommending practices for disposal based on empirical, quantitative evidence.

Original languageEnglish (US)
Article numbere0202294
JournalPLoS One
Volume13
Issue number8
DOIs
StatePublished - Aug 1 2018

Fingerprint

homoserine
lactones
Escherichia coli
quorum sensing
Quorum Sensing
Crosstalk
Bioluminescence
group behavior
bioluminescence
autoclaving
Chassis
Biofilms
biofilm
Libraries
Agar
Virulence
homoserine lactone
virulence
agar
Molecules

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Daer, R., Barrett, C. M., Melendez, E. L., Wu, J., Tekel, S. J., Xu, J., ... Haynes, K. (2018). Characterization of diverse homoserine lactone synthases in Escherichia coli. PLoS One, 13(8), [e0202294]. https://doi.org/10.1371/journal.pone.0202294

Characterization of diverse homoserine lactone synthases in Escherichia coli. / Daer, René; Barrett, Cassandra M.; Melendez, Ernesto Luna; Wu, Jiaqi; Tekel, Stefan J.; Xu, Jimmy; Dennison, Brady; Muller, Ryan; Haynes, Karmella.

In: PLoS One, Vol. 13, No. 8, e0202294, 01.08.2018.

Research output: Contribution to journalArticle

Daer, R, Barrett, CM, Melendez, EL, Wu, J, Tekel, SJ, Xu, J, Dennison, B, Muller, R & Haynes, K 2018, 'Characterization of diverse homoserine lactone synthases in Escherichia coli', PLoS One, vol. 13, no. 8, e0202294. https://doi.org/10.1371/journal.pone.0202294
Daer R, Barrett CM, Melendez EL, Wu J, Tekel SJ, Xu J et al. Characterization of diverse homoserine lactone synthases in Escherichia coli. PLoS One. 2018 Aug 1;13(8). e0202294. https://doi.org/10.1371/journal.pone.0202294
Daer, René ; Barrett, Cassandra M. ; Melendez, Ernesto Luna ; Wu, Jiaqi ; Tekel, Stefan J. ; Xu, Jimmy ; Dennison, Brady ; Muller, Ryan ; Haynes, Karmella. / Characterization of diverse homoserine lactone synthases in Escherichia coli. In: PLoS One. 2018 ; Vol. 13, No. 8.
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