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Targeted Repression of Essential Genes To Arrest Growth and Increase Carbon Partitioning and Biofuel Titers in Cyanobacteria
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Systems Biology.
KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0002-2430-2682
KTH.
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Systems Biology.
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2018 (English)In: ACS Synthetic Biology, E-ISSN 2161-5063, Vol. 7, no 7, article id diva2:1239079Article in journal, Letter (Refereed) Published
Abstract [en]

Photoautotrophic production of fuels and chemicals by cyanobacteria typically gives lower volumetric productivities and titers than heterotrophic production. Cyanobacteria cultures become light limited above an optimal cell density, so that this substrate is not supplied to all cells sufficiently. Here, we investigate genetic strategies for a two-phase cultivation, where biofuel-producing Synechocystis cultures are limited to an optimal cell density through inducible CRISPR interference (CRISPRi) repression of cell growth. Fixed CO2 is diverted to ethanol or n-butanol. Among the most successful strategies was partial repression of citrate synthase gltA. Strong repression (>90%) of gitA at low culture densities increased carbon partitioning to n-butanol 5-fold relative to a nonrepression strain, but sacrificed volumetric productivity due to severe growth restriction. CO2 fixation continued for at least 3 days after growth was arrested. By targeting sgRNAs to different regions of the gitA gene, we could modulate GItA expression and carbon partitioning between growth and product to increase both specific and volumetric productivity. These growth arrest strategies can be useful for improving performance of other photoautotrophic processes.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2018. Vol. 7, no 7, article id diva2:1239079
Keywords [en]
cyanobacteria, CRISPRi, bioproduction
National Category
Biochemistry and Molecular Biology
Research subject
Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-235174DOI: 10.1021/acssynbio.8b00056OAI: oai:DiVA.org:kth-235174DiVA, id: diva2:1248671
Funder
EU, Horizon 2020, 760994
Note

QC 20180920

Available from: 2018-09-17 Created: 2018-09-17 Last updated: 2018-09-20Bibliographically approved

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Shabestary, KiyanAnfelt, JosefinYao, LunHudson, Elton P.

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Shabestary, KiyanAnfelt, JosefinLjungqvist, EmilJahn, MichaelYao, LunHudson, Elton P.
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