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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.
Biotechnology for biofuels 2015 v.8 no.1 pp. 193
DNA, Firmicutes, Methanoculleus, Proteobacteria, Spirochaetales, bacteria, biogas, bioinformatics, biomass, biotechnology, data collection, enzymes, fermentation, genes, high-throughput nucleotide sequencing, metagenomics, methane, methane production, methanogens, microbial communities, ribosomal RNA
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.