title: Culturing Synechocystis sp. Strain PCC 6803 with N2 and CO2 in a Diel Regime Reveals Multiphase Glycogen Dynamics with Low Maintenance Costs.
abstract: Investigating the physiology of cyanobacteria cultured under a diel light regime is relevant for a better understanding of the resulting growth characteristics and for specific biotechnological applications that are foreseen for these photosynthetic organisms. Here, we present the results of a multiomics study of the model cyanobacterium Synechocystis sp. strain PCC 6803, cultured in a lab-scale photobioreactor in physiological conditions relevant for large-scale culturing. The culture was sparged with N2 and CO2, leading to an anoxic environment during the dark period. Growth followed the availability of light. Metabolite analysis performed with (1)H nuclear magnetic resonance analysis showed that amino acids involved in nitrogen and sulfur assimilation showed elevated levels in the light. Most protein levels, analyzed through mass spectrometry, remained rather stable. However, several high-light-response proteins and stress-response proteins showed distinct changes at the onset of the light period. Microarray-based transcript analysis found common patterns of ∼56% of the transcriptome following the diel regime. These oscillating transcripts could be grouped coarsely into genes that were upregulated and downregulated in the dark period. The accumulated glycogen was degraded in the anaerobic environment in the dark. A small part was degraded gradually, reflecting basic maintenance requirements of the cells in darkness. Surprisingly, the largest part was degraded rapidly in a short time span at the end of the dark period. This degradation could allow rapid formation of metabolic intermediates at the end of the dark period, preparing the cells for the resumption of growth at the start of the light period.
  
  
  
  Industrial-scale biotechnological applications are anticipated for cyanobacteria. We simulated large-scale high-cell-density culturing of Synechocystis sp. PCC 6803 under a diel light regime in a lab-scale photobioreactor. In BG-11 medium, Synechocystis grew only in the light. Metabolite analysis grouped the collected samples according to the light and dark conditions. Proteome analysis suggested that the majority of enzyme-activity regulation was not hierarchical but rather occurred through enzyme activity regulation. An abrupt light-on condition induced high-light-stress proteins. Transcript analysis showed distinct patterns for the light and dark periods. Glycogen gradually accumulated in the light and was rapidly consumed in the last quarter of the dark period. This suggests that the circadian clock primed the cellular machinery for immediate resumption of growth in the light.
authors.affiliation: Molecular Microbial Physiology, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: S Andreas
authors.initials: SA
authors.lastname: Angermayr
authors.affiliation: Molecular Microbial Physiology, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Pascal
authors.initials: P
authors.lastname: van Alphen
authors.affiliation: Biosystems Data Analysis, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Dicle
authors.initials: D
authors.lastname: Hasdemir
authors.affiliation: Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Gertjan
authors.initials: G
authors.lastname: Kramer
authors.affiliation: Natural Products Laboratory, Institute of Biology, Leiden University, Leiden, The Netherlands.
authors.forename: Muzamal
authors.initials: M
authors.lastname: Iqbal
authors.affiliation: Photanol B.V., Amsterdam, The Netherlands.
authors.forename: Wilmar
authors.initials: W
authors.lastname: van Grondelle
authors.affiliation: Biosystems Data Analysis, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Huub C
authors.initials: HC
authors.lastname: Hoefsloot
authors.affiliation: Natural Products Laboratory, Institute of Biology, Leiden University, Leiden, The Netherlands.
authors.forename: Young Hae
authors.initials: YH
authors.lastname: Choi
authors.affiliation: Molecular Microbial Physiology, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands k.j.hellingwerf@uva.nl.|Photanol B.V., Amsterdam, The Netherlands.
authors.forename: Klaas J
authors.initials: KJ
authors.lastname: Hellingwerf
mesh_terms: D000332:Aerobiosis; D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000032:analysis; D002245:Carbon Dioxide / Q000378:metabolism*; D003624:Darkness; D020869:Gene Expression Profiling; D006003:Glycogen / Q000378:metabolism*; D008027:Light; D013058:Mass Spectrometry; D046228:Microarray Analysis; D009584:Nitrogen / Q000378:metabolism*; D058433:Photobioreactors / Q000382:microbiology; D046939:Synechocystis / Q000737:chemistry / Q000254:growth & development* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D002245:Carbon Dioxide; D006003:Glycogen; D009584:Nitrogen