TY - JOUR
T1 - Characterization of diverse homoserine lactone synthases in Escherichia coli
AU - Daer, René
AU - Barrett, Cassandra M.
AU - Melendez, Ernesto Luna
AU - Wu, Jiaqi
AU - Tekel, Stefan J.
AU - Xu, Jimmy
AU - Dennison, Brady
AU - Muller, Ryan
AU - Haynes, Karmella
N1 - Funding Information:
Most of the work reported here was presented at the 2016 International Genetically Engineered Machines (iGEM) Competition by the Arizona State University (ASU) 2016 iGEM team. The authors thank Integrated DNA Technologies (IDT, http://idtdna.com), iGEM Headquarters (http://igem.org), the Western Alliance to Expand Student Opportunities (WAESO, NSF HRD 1401190, https://mge-p1000.asu.edu/waeso/), the School of Molecular Sciences at Arizona State University, and the School of Biological and Health Systems Engineering at Arizona State University for their support of the undergraduate co-authors. KAH was supported by the NIH NCI (K01CA188164, http://www.nih.gov). RMD was supported by the ARCS Foundation (https://www.arcsfoundation.org/) and NSF CBET (1404084, http://nsf.gov). CMB is supported by the Ira A. Fulton School of Engineering at Arizona State University. Most of the work reported here was presented at the 2016 International Genetically Engineered Machines (iGEM) Competition by the Arizona State University (ASU) 2016 iGEM team. The authors thank J. Steel (DNASU) for DNA sequencing services. The authors thank R. Allen, S. Raman, G. Srinivasan and J. Daer for critical review of the manuscript. The authors thank Integrated DNA Technologies (IDT, www.idtdna.com), iGEM Headquarters (www.igem.org), the Western Alliance to Expand Student Opportunities (WAESO, NSF HRD 1401190, https://mge-p1000.asu.edu/waeso/), the School of Molecular Sciences at Arizona State University, and the School of Biological and Health Systems Engineering at Arizona State University for their support of the undergraduate co-authors. KAH was supported by the NIH NCI (K01CA188164, www.nih.gov). RMD was supported by the ARCS Foundation (www.arcsfoundation.org) and NSF CBET (1404084, www.nsf.gov). CMB is supported by the Ira A. Fulton School of Engineering at Arizona State University.
Publisher Copyright:
© 2018 Daer et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2018/8
Y1 - 2018/8
N2 - 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.
AB - 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.
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U2 - 10.1371/journal.pone.0202294
DO - 10.1371/journal.pone.0202294
M3 - Article
C2 - 30138364
AN - SCOPUS:85052146165
SN - 1932-6203
VL - 13
JO - PloS one
JF - PloS one
IS - 8
M1 - e0202294
ER -