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

Hammar, Petter and Angermayr, Andreas S and Sjostrom , Staffan L and Van Der Meer , Josefin and Hellingwerf , Klaas J and Hudson , Elton P and Joensson, Haakan N (2015) Single-cell screening of photosynthetic growth and lactate production by cyanobacteria. Biotechnology for Biofuels, 8. Article No. 193. ISSN 17546834

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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.

Item Type: Article
DOI: 10.1186/s13068-015-0380-2
Uncontrolled Keywords: Single cell, Cyanobacteria, Droplet microfluidics, High-throughput screening, Lactate dehydrogenase
Subjects: 500 Science > 570 Life sciences; biology
Research Group: Bollenbach Group
SWORD Depositor: Sword Import User
Depositing User: Sword Import User
Date Deposited: 15 Jan 2016 13:05
Last Modified: 30 Aug 2017 10:56

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