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Single-cell screening of photosynthetic growth and lactate production by cyanobacteria

BACKGROUND: Photosynthetic cyanobacteria are attractive for a range of biotechnological applications including biofuel production. However, due to slow growth, screening of mutant libraries using microtiter plates is not feasible. RESULTS: We present a method for high-throughput, single-cell analysi...

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Autores principales: Hammar, Petter, Angermayr, S. Andreas, Sjostrom, Staffan L., van der Meer, Josefin, Hellingwerf, Klaas J., Hudson, Elton P., Joensson, Haakan N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4660834/
https://www.ncbi.nlm.nih.gov/pubmed/26613003
http://dx.doi.org/10.1186/s13068-015-0380-2
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author Hammar, Petter
Angermayr, S. Andreas
Sjostrom, Staffan L.
van der Meer, Josefin
Hellingwerf, Klaas J.
Hudson, Elton P.
Joensson, Haakan N.
author_facet Hammar, Petter
Angermayr, S. Andreas
Sjostrom, Staffan L.
van der Meer, Josefin
Hellingwerf, Klaas J.
Hudson, Elton P.
Joensson, Haakan N.
author_sort Hammar, Petter
collection PubMed
description BACKGROUND: Photosynthetic cyanobacteria are attractive for a range of biotechnological applications including biofuel production. However, due to slow growth, screening of mutant libraries using microtiter plates is not feasible. RESULTS: We present a method for high-throughput, single-cell analysis and sorting of genetically engineered l-lactate-producing strains of Synechocystis sp. PCC6803. A microfluidic device is used to encapsulate single cells in picoliter droplets, assay the droplets for l-lactate production, and sort strains with high productivity. We demonstrate the separation of low- and high-producing reference strains, as well as enrichment of a more productive l-lactate-synthesizing population after UV-induced mutagenesis. The droplet platform also revealed population heterogeneity in photosynthetic growth and lactate production, as well as the presence of metabolically stalled cells. CONCLUSIONS: The workflow will facilitate metabolic engineering and directed evolution studies and will be useful in studies of cyanobacteria biochemistry and physiology. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-015-0380-2) contains supplementary material, which is available to authorized users.
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spelling pubmed-46608342015-11-27 Single-cell screening of photosynthetic growth and lactate production by cyanobacteria Hammar, Petter Angermayr, S. Andreas Sjostrom, Staffan L. van der Meer, Josefin Hellingwerf, Klaas J. Hudson, Elton P. Joensson, Haakan N. Biotechnol Biofuels Methodology BACKGROUND: Photosynthetic cyanobacteria are attractive for a range of biotechnological applications including biofuel production. However, due to slow growth, screening of mutant libraries using microtiter plates is not feasible. RESULTS: We present a method for high-throughput, single-cell analysis and sorting of genetically engineered l-lactate-producing strains of Synechocystis sp. PCC6803. A microfluidic device is used to encapsulate single cells in picoliter droplets, assay the droplets for l-lactate production, and sort strains with high productivity. We demonstrate the separation of low- and high-producing reference strains, as well as enrichment of a more productive l-lactate-synthesizing population after UV-induced mutagenesis. The droplet platform also revealed population heterogeneity in photosynthetic growth and lactate production, as well as the presence of metabolically stalled cells. CONCLUSIONS: The workflow will facilitate metabolic engineering and directed evolution studies and will be useful in studies of cyanobacteria biochemistry and physiology. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-015-0380-2) contains supplementary material, which is available to authorized users. BioMed Central 2015-11-25 /pmc/articles/PMC4660834/ /pubmed/26613003 http://dx.doi.org/10.1186/s13068-015-0380-2 Text en © Hammar et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology
Hammar, Petter
Angermayr, S. Andreas
Sjostrom, Staffan L.
van der Meer, Josefin
Hellingwerf, Klaas J.
Hudson, Elton P.
Joensson, Haakan N.
Single-cell screening of photosynthetic growth and lactate production by cyanobacteria
title Single-cell screening of photosynthetic growth and lactate production by cyanobacteria
title_full Single-cell screening of photosynthetic growth and lactate production by cyanobacteria
title_fullStr Single-cell screening of photosynthetic growth and lactate production by cyanobacteria
title_full_unstemmed Single-cell screening of photosynthetic growth and lactate production by cyanobacteria
title_short Single-cell screening of photosynthetic growth and lactate production by cyanobacteria
title_sort single-cell screening of photosynthetic growth and lactate production by cyanobacteria
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4660834/
https://www.ncbi.nlm.nih.gov/pubmed/26613003
http://dx.doi.org/10.1186/s13068-015-0380-2
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