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Title: Synthetic photosynthetic consortia define interactions leading to robustness and photoproduction

In this study, microbial consortia composed of autotrophic and heterotrophic species abound in nature, yet examples of synthetic communities with mixed metabolism are limited in the laboratory. We previously engineered a model cyanobacterium, Synechococcus elongatus PCC 7942, to secrete the bulk of the carbon it fixes as sucrose, a carbohydrate that can be utilized by many other microbes. Here, we tested the capability of sucrose-secreting cyanobacteria to act as a flexible platform for the construction of synthetic, light-driven consortia by pairing them with three disparate heterotrophs: Bacillus subtilis, Escherichia coli, or Saccharomyces cerevisiae. The comparison of these different co-culture dyads reveals general design principles for the construction of robust autotroph/heterotroph consortia. As a result, we observed heterotrophic growth dependent upon cyanobacterial photosynthate in each co-culture pair. Furthermore, these synthetic consortia could be stabilized over the long-term (weeks to months) and both species could persist when challenged with specific perturbations. Stability and productivity of autotroph/heterotroph co-cultures was dependent on heterotroph sucrose utilization, as well as other species-independent interactions that we observed across all dyads. One destabilizing interaction we observed was that non-sucrose byproducts of oxygenic photosynthesis negatively impacted heterotroph growth. Conversely, inoculation of each heterotrophic species enhanced cyanobacterial growth in comparisonmore » to axenic cultures. Finally, these consortia can be flexibly programmed for photoproduction of target compounds and proteins; by changing the heterotroph in co-culture to specialized strains of B. subtilis or E. coli we demonstrate production of alpha-amylase and polyhydroxybutyrate, respectively. In conclusion, enabled by the unprecedented flexibility of this consortia design, we uncover species-independent design principles that influence cyanobacteria/heterotroph consortia robustness. The modular nature of these communities and their unusual robustness exhibits promise as a platform for highly-versatile photoproduction strategies that capitalize on multi-species interactions and could be utilized as a tool for the study of nascent symbioses. Further consortia improvements via engineered interventions beyond those we show here (i.e., increased efficiency growing on sucrose) could improve these communities as production platforms.« less
Authors:
 [1] ;  [2] ;  [1] ;  [3]
  1. Harvard Medical School, Boston, MA (United States); Harvard Univ., Boston, MA (United States)
  2. Michigan State Univ., East Lansing, MI (United States); Washington Univ., St. Louis, MO (United States)
  3. Michigan State Univ., East Lansing, MI (United States)
Publication Date:
Grant/Contract Number:
SC0012658
Type:
Accepted Manuscript
Journal Name:
Journal of Biological Engineering
Additional Journal Information:
Journal Volume: 11; Journal Issue: 1; Journal ID: ISSN 1754-1611
Publisher:
BioMed Central
Research Org:
Johns Hopkins Univ., Baltimore, MD (United States)
Sponsoring Org:
USDOE Office of Science (SC)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; synthetic biology; photoproduction; synthetic consortia; microbial communities
OSTI Identifier:
1367177

Hays, Stephanie G., Yan, Leo L. W., Silver, Pamela A., and Ducat, Daniel C.. Synthetic photosynthetic consortia define interactions leading to robustness and photoproduction. United States: N. p., Web. doi:10.1186/s13036-017-0048-5.
Hays, Stephanie G., Yan, Leo L. W., Silver, Pamela A., & Ducat, Daniel C.. Synthetic photosynthetic consortia define interactions leading to robustness and photoproduction. United States. doi:10.1186/s13036-017-0048-5.
Hays, Stephanie G., Yan, Leo L. W., Silver, Pamela A., and Ducat, Daniel C.. 2017. "Synthetic photosynthetic consortia define interactions leading to robustness and photoproduction". United States. doi:10.1186/s13036-017-0048-5. https://www.osti.gov/servlets/purl/1367177.
@article{osti_1367177,
title = {Synthetic photosynthetic consortia define interactions leading to robustness and photoproduction},
author = {Hays, Stephanie G. and Yan, Leo L. W. and Silver, Pamela A. and Ducat, Daniel C.},
abstractNote = {In this study, microbial consortia composed of autotrophic and heterotrophic species abound in nature, yet examples of synthetic communities with mixed metabolism are limited in the laboratory. We previously engineered a model cyanobacterium, Synechococcus elongatus PCC 7942, to secrete the bulk of the carbon it fixes as sucrose, a carbohydrate that can be utilized by many other microbes. Here, we tested the capability of sucrose-secreting cyanobacteria to act as a flexible platform for the construction of synthetic, light-driven consortia by pairing them with three disparate heterotrophs: Bacillus subtilis, Escherichia coli, or Saccharomyces cerevisiae. The comparison of these different co-culture dyads reveals general design principles for the construction of robust autotroph/heterotroph consortia. As a result, we observed heterotrophic growth dependent upon cyanobacterial photosynthate in each co-culture pair. Furthermore, these synthetic consortia could be stabilized over the long-term (weeks to months) and both species could persist when challenged with specific perturbations. Stability and productivity of autotroph/heterotroph co-cultures was dependent on heterotroph sucrose utilization, as well as other species-independent interactions that we observed across all dyads. One destabilizing interaction we observed was that non-sucrose byproducts of oxygenic photosynthesis negatively impacted heterotroph growth. Conversely, inoculation of each heterotrophic species enhanced cyanobacterial growth in comparison to axenic cultures. Finally, these consortia can be flexibly programmed for photoproduction of target compounds and proteins; by changing the heterotroph in co-culture to specialized strains of B. subtilis or E. coli we demonstrate production of alpha-amylase and polyhydroxybutyrate, respectively. In conclusion, enabled by the unprecedented flexibility of this consortia design, we uncover species-independent design principles that influence cyanobacteria/heterotroph consortia robustness. The modular nature of these communities and their unusual robustness exhibits promise as a platform for highly-versatile photoproduction strategies that capitalize on multi-species interactions and could be utilized as a tool for the study of nascent symbioses. Further consortia improvements via engineered interventions beyond those we show here (i.e., increased efficiency growing on sucrose) could improve these communities as production platforms.},
doi = {10.1186/s13036-017-0048-5},
journal = {Journal of Biological Engineering},
number = 1,
volume = 11,
place = {United States},
year = {2017},
month = {1}
}

Works referenced in this record:

One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products
journal, May 2000
  • Datsenko, K. A.; Wanner, B. L.
  • Proceedings of the National Academy of Sciences, Vol. 97, Issue 12, p. 6640-6645
  • DOI: 10.1073/pnas.120163297