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Cyanobacterial circadian regulation enhances bioproduction under subjective nighttime through rewiring of carbon partitioning dynamics, redox balance orchestration, and cell cycle modulation

ABSTRACT: GC-MS extracellular metabolite profiling of circadian clock regulation impacts of carbon partitioning between storage, growth, and product synthesis in Synechococcus elongatus PCC 7942 for enhanced bioproduction strategy investigation. Metabolomic targeted analysis of PCC 7942 light-dark cycle cultures transitioned to constant light revealed distinct temporal patterns in sucrose production. Culture samples (8 mL) were collected at 0, 0.5, 1, 2, 4, 6, and 8 hours then dried under speed vacuum and dissolved in pyridine and chemically derivatized by methoxyamination and trimethylsilylation for extracellular sucrose analysis. Data was analyzed using Agilent Mass Hunter. Culture samples (8 mL) were collected at 0, 0.5, 1, 2, 4, 6, and 8 hours then dried under speed vacuum and dissolved in pyridine and chemically derivatized by methoxyamination and trimethylsilylation for extracellular sucrose analysis.

INSTRUMENT(S): 8890 GC/MS

ORGANISM(S): Synechococcus Elongatus Pcc 7942 (ncbitaxon:1140)

SUBMITTER: Pavlo Bohutskyi

PROVIDER: MSV000096722 | MassIVE | Sat Dec 21 20:48:00 GMT 2024

REPOSITORIES: MassIVE

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Cyanobacterial circadian regulation enhances bioproduction under subjective nighttime through rewiring of carbon partitioning dynamics, redox balance orchestration, and cell cycle modulation.

Gilliam Ashley A Sadler Natalie C NC Li Xiaolu X Garcia Marci M Johnson Zachary Z Veličković Marija M Kim Young-Mo YM Feng Song S Qian Wei-Jun WJ Cheung Margaret S MS Bohutskyi Pavlo P

Microbial cell factories 20250308 1

<h4>Background</h4>The industrial feasibility of photosynthetic bioproduction using cyanobacterial platforms remains challenging due to insufficient yields, particularly due to competition between product formation and cellular carbon demands across different temporal phases of growth. This study investigates how circadian clock regulation impacts carbon partitioning between storage, growth, and product synthesis in Synechococcus elongatus PCC 7942, and provides insights that suggest potential s ...[more]

PMID: 40055679

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Project description:In the unicellular cyanobacterium, Synechococcus elongatus PCC 7942, essentially all promoter activities are under the control of the circadian clock in continuous light (LL) conditions. Here, we employed high-density oligonucleotide arrays to explore comprehensive profiles of genome-wide Synechococcus gene expression in wild type, kaiABC-null and kaiC-overexpressor strains under LL and continuous dark (DD) conditions. In the wild type strain more than 30% of transcripts significantly oscillated in a circadian fashion, peaking at subjective dawn and dusk. Such circadian control was nullified in kaiABC-null strains. Although KaiC has been proposed to globally repress gene expression, our analysis revealed that dawn expressing genes were upregulated by kaiC-overexpression, such that the clock was arrested at subjective dawn. Transfer of cells to continuous dark (DD) conditions from LL immediately suppressed expression of most of genes, while the clock keeps time even in the absence of transcriptional feedback. Thus, the Synechococcus genome seems primarily regulated by the light/dark cycles and dramatically modified by the protein-based circadian oscillator. Keywords: timecourse data (~48 hours under continuous light or darkness) from Synechococcus elongatus PCC 7942 (wild type, kaiABC-null, and inducible kaiC-overexpressor) strains Wild type (WT), kaiABC-null (Dkai), and inducible kaiC-overexpressor (oxC) S. elongatus PCC 7942 strains were analyzed under continuous light (LL), continuous dark (DD) using Affymetrix high-density oligonucleotide microarrays (GeneChip CustomExpress Arrays) representing predicted 2,515 protein-coding genes on the genome of Synechococcus elongatus PCC 6301, which can be used also to the almost homologous strain, S. elongatus PCC 7942: Two independent experiments in WT under LL (hours 0 to 52) and DD (hours 0 to 48); Two independent experiments in Dkai under LL (hours 0 to 48); Two independent experiments in oxC under LL in the presence or absence of an inducer, IPTG, from hour 25 to 33 in LL; a single experiment in WT under DD in the presence of an transcriptional inhibitor, rifampicin.