Genetically Programmable Microbial Assembly
Abstract
Engineered microbial communities show promise in a wide range of applications, including environmental remediation, microbiome engineering, and synthesis of fine chemicals. Here we present methods by which bacterial aggregates can be directed into several distinct architectures by inducible surface expression of heteroassociative protein domains (SpyTag/SpyCatcher and SynZip17/18). Programmed aggregation can be used to activate a quorum-sensing circuit, and aggregate size can be tuned via control of the amount of the associative protein displayed on the cell surface. We further demonstrate reversibility of SynZip-mediated assembly by addition of soluble competitor peptide. Genetically programmable bacterial assembly provides a starting point for the development of new applications of engineered microbial communities in environmental technology, agriculture, human health, and bioreactor design.
Additional Information
© 2021 American Chemical Society. Received: December 7, 2020; Published: May 19, 2021. This work was supported by the Defense Advanced Research Projects Agency under award numbers HR0011-15-C-0093 and HR0011-17-2-0037. M.T.K. was supported by the Department of Defense (DoD) through the National Defense Science & Engineering Graduate Fellowship (NDSEG) Program. B.R.S. was supported by NIH Training Grant 1T32GM112592. C.P.J. was supported by a Caltech Summer Undergraduate Research Fellowship. Plasmid pHEA-Cm, from which the autotransporter was cloned, was a kind gift of Luis Angel Fernandez Herrero of the Centro Nacional de Biotecnologia (Madrid, Spain). Plasmid pLuxRI2, from which we cloned the quorum-sensing plasmid, was a gift of Frances Arnold of the California Institute of Technology (Pasadena, California). pKPY680, pKPY681, and DH10B strains KY35 and KY36 were prepared by Kai P. Yuet. We thank Dr. Andres Collazo for assistance with confocal microscopy. Imaging was performed in the Biological Imaging Facility of the Caltech Beckman Institute, which is supported by the Arnold and Mabel Beckman Foundation. We thank Yaron Antebi for use of EasyFlow for flow cytometry analysis, and Adam Silverman for a critical review of the manuscript. Author Contributions: M.T.K. and B.R.S. contributed equally to this work. Author Contributions: M.T.K., B.R.S., and D.A.T. conceived the study. M.T.K., B.R.S., and C.P.J. conducted all the experiments in the paper. B.R.S. wrote computational tools for image analysis and reaction diffusion modeling. M.T.K., B.R.S., and D.A.T. wrote the paper. All authors read and approved the final manuscript. The authors declare no competing financial interest.Attached Files
Accepted Version - nihms-1789073.pdf
Supplemental Material - sb0c00616_si_001.pdf
Supplemental Material - sb0c00616_si_002.zip
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Additional details
- PMCID
- PMC8978479
- Eprint ID
- 109192
- Resolver ID
- CaltechAUTHORS:20210519-141320901
- HR0011-15-C-0093
- Defense Advanced Research Projects Agency (DARPA)
- HR0011-17-2-0037
- Defense Advanced Research Projects Agency (DARPA)
- National Defense Science and Engineering Graduate (NDSEG) Fellowship
- 1T32GM112592
- NIH Predoctoral Fellowship
- National Institute of General Medical Sciences
- Caltech Summer Undergraduate Research Fellowship (SURF)
- Arnold and Mabel Beckman Foundation
- Created
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2021-05-24Created from EPrint's datestamp field
- Updated
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2022-07-12Created from EPrint's last_modified field