Skip to main content
SpringerLink
Log in

Methods for Generating Microbial Cocultures that Grow in the Absence of Fixed Carbon or Nitrogen

  • Protocol
  • First Online:
Synthetic Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1772))

Abstract

This work is motivated by both (1) the desire to make interesting products without the reliance on fixed carbon or fixed nitrogen using microbial cocultures, and (2) the desire to develop new methods for growing microbial communities. The bioplastic polyhydroxybutyrate (PHB), for example, is considered prohibitively expensive to make from sugar (compared to polypropylene). Utilizing and building on engineered strains of Synechococcus elongatus and Azotobacter vinelandii, we have combined a nitrogen-fixing organism with a carbon-fixing organism to make PHB from air, water, sunlight, and trace minerals. Our observations of coculture growth in batch culture led us to develop an improved system based on manipulating the osmotic pressure within a hydrogel. The methods we used to develop this coculture are described in detail in the following chapter, including notes detailing some of our additional observations or thoughts on this system.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 279.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Traxler MF, Watrous JD, Alexandrov T, Dorrestein PC, Kolter R (2013) Interspecies interactions stimulate diversification of the Streptomyces coelicolor secreted metabolome. MBio 4(4). https://doi.org/10.1128/mBio.00459-13

    Article  Google Scholar 

  2. Bader J, Mast-Gerlach E, Popovic M, Baipai R, Stahl U (2010) Relevance of microbial coculture fermentations in biotechnology. J Appl Microbiol 109:371–387

    Article  CAS  Google Scholar 

  3. Goers L, Freemont P, Polizzi KM (2014) Co-culture systems and technologies: taking synthetic biology to the next level. J R Soc Interface 11:20140065

    Article  Google Scholar 

  4. Smith MJ, Francis MB (2016) A designed A. vinelandii-S. elongatus coculture for chemical photoproduction from air, water, phosphate and trace metals. ACS Synth Biol 5(9):955–961

    Article  CAS  Google Scholar 

  5. Setubal JC et al (2009) Genome sequence of Azotobacter vinelandii, an obligate aerobe specialized to support diverse anaerobic metabolic processes. J Bacteriol 191:4534–4545

    Article  CAS  Google Scholar 

  6. El-Shanshoury AE, Kenawy E, Amara AA, Salama AF, Kishk SS (2013) Optimization of Polyhydroxybutyrate (PHB) production by Azotobacter vinelandii using experimental design. Int J Curr Microbiol Appl Sci 2:227–241

    Google Scholar 

  7. Brewin B, Woodley P, Drummond M (1999) The basis of ammonium release in nifL mutants of Azotobacter vinelandii. J Bacteriol 181(23):7356–7362

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Ortiz-Marquez JC, Nascimento MD, Dublan M, Curatti L (2012) Association with an ammonium-excreting bacterium allows diazotrophic culture of oil-rich eukaryotic microalgae. Appl Environ Microbiol 78:2345–2352

    Article  CAS  Google Scholar 

  9. Ducat DC, Avelar-Rivas JA, Way JC, Silver PA (2012) Rerouting carbon flux to enhance photosynthetic productivity. Appl Environ Microbiol 78:2660–2668

    Article  CAS  Google Scholar 

  10. Smith MJ, Francis MB (2017) Improving metabolite production in microbial cocultures using a spatially-constrained hydrogel. Biotechnol Bioeng 114(6):1195–1200

    Article  CAS  Google Scholar 

  11. Stanier R, Kunisawa R, Mandel M, Cohen-Bazire G (1971) Purification and properties of unicellular blue-green algae (order Cchroococcales). Bacteriol Rev 35:171–205

    CAS  PubMed  PubMed Central  Google Scholar 

  12. D’Mello R, Hill S, Poole RK (1994) Determination of the oxygen affinities of terminal oxidases in Azotobacter vinelandii using the deoxygenation of oxyleghaemoglobin and oxymyoglobin: cytochrome bd is a low-affinity oxidase. Microbiology 140:1395–1402

    Article  Google Scholar 

  13. Engler C, Kandzia R, Marillonnet S (2008) A one pot, one step, precision cloning method with high throughput capability. PLoS One 3:e3647

    Article  Google Scholar 

  14. Porra RJ (2002) The chequered history of the development and use of simultaneous equations for the accurate determination of chlorophylls a and b. Photosynth Res 73:149–156

    Article  CAS  Google Scholar 

  15. Law JH, Slepecky RA (1961) Assay of poly-β-hydroxybutyric acid. J Bacteriol 82:33–36

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Chemistry, University of California, Berkeley, CA, USA

    Matthew J. Smith

  2. Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA

    Matthew B. Francis

Authors
  1. Matthew J. Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Matthew B. Francis
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Matthew B. Francis .

Editor information

Editors and Affiliations

  1. Agilent Technologies, Inc, La Jolla, California, USA

    Jeffrey Carl Braman

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Smith, M.J., Francis, M.B. (2018). Methods for Generating Microbial Cocultures that Grow in the Absence of Fixed Carbon or Nitrogen. In: Braman, J. (eds) Synthetic Biology. Methods in Molecular Biology, vol 1772. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7795-6_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7795-6_3

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7794-9

  • Online ISBN: 978-1-4939-7795-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation