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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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Detalles Bibliográficos
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
Descripción
Sumario: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.