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

Author:
Hammar, Petter, Angermayr, S. Andreas, Sjostrom, Staffan L., van der Meer, Josefin, Hellingwerf, Klaas J., Hudson, Elton P., Joensson, Haakan N.
Source:
Biotechnology for biofuels 2015 v.8 no.1 pp. 193
ISSN:
1754-6834
Subject:
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
Abstract:
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.
Agid:
1212715