Project description:We installed and optimized a genetic tool that allows control over protein stability in a model cyanobacterium. This tool has potential uses for the fundamental study of cyanobacterial genes, and may be useful for the design of more sophisticated, bioindustrial cyanobacterial strains.
Project description:The study aims to identify the metabolic differences between two promising fast-growing, non-model cyanobacterial strains, S. elongatus PCC 11801 and PCC 11802. To this end, experiments were carried out to measure metabolite levels in the two cyanobacterial strains grown in shake flasks at a similar light intensity of approx. 300-350 µmole photons.m-2. s-1. The samples for metabolomics analysis were collected during the exponential growth phase at an optical cell density of 0.5-0.6. Isotopic ratio method was utilized to compare the metabolite levels and delineate the differences in their metabolic pathways.
Project description:The catalytic core of the RNA polymerase of most eubacteria is composed of two α subunits and β, β’ and ω subunits. In Escherichia coli, the ω subunit (encoded by the rpoZ gene) has been suggested to assist β’ during RNA polymerase core assembly. The function of the ω subunit is particularly interesting in cyanobacteria because the cyanobacterial β’ is split to N-terminal γ and C-terminal β’ subunits. The ∆rpoZ strain of the cyanobacterium Synechocystis sp. PCC 6803 grew well in standard conditions although the mutant cells showed low light-saturated photosynthetic activity, low Rubisco content and accumulated high quantities of protective carotenoids and α-tocopherol. The ∆rpoZ strain contained 15% less of the primary σ factor, SigA, than the control strain, and recruitment of SigA to the RNA polymerase core was inefficient in ∆rpoZ. Thus, a cyanobacterial RNA polymerase holoenzyme lacking the ω subunit contains less frequently the primary σ factor. A DNA microarray analysis revealed that this leads to specific down-regulation of highly expressed genes, like genes encoding subunits for Rubisco, ATP synthase, NADH-dehydrogenase and carbon concentrating mechanisms. On the contrary, many genes showing only low or moderate expression in the control strain were up-regulated in ∆rpoZ. A conserved -10 region was detected in promoters showing up or down-regulation in ∆rpoZ, but -35 regions of down-regulated genes completely differed from -35 regions of up-regulated genes.
Project description:The study aims to identify the metabolic differences between two promising fast-growing, non-model cyanobacterial strains, S. elongatus PCC 11801 and PCC 11802. To this end, dynamic 13C-labeling experiments were carried out in the two cyanobacterial strains grown in shake flasks at a similar light intensity of approx. 300-350 µmole photons.m-2. s-1. The samples for metabolomics analysis were collected during the exponential growth phase at an optical cell density of 0.5-0.6. The detailed protocol for experiment can be found in the protocol file.
Project description:The NDH1 complex fulfils numerous tasks in the cyanobacterial cell such as respiration, cyclic electron flow, and inorganic carbon concentration. Despite the immense progress in our understanding of structure/function relation of the cyanobacterial NDH1 complex, the subunits catalysing the NAD(P)H docking and oxidation are still missing. The gene sml0013 of Synechocystis 6803 encodes for a small protein of unknown function for that homologs exist in all completely known cyanobacterial genomes. The protein exhibits weak similarities to the NDF6 protein, which was reported from Arabidopsis chloroplasts as a NDH subunit (Ishikawa et al. 2008). A sml0013 inactivation mutant of Synechocystis 6803 was generated and characterized. It showed only weak differences regarding growth and pigmentation at various culture conditions; most remarkably it exhibited a glucose-sensitive phenotype in the light. The genome-wide expression pattern of the Δsml0013::Km mutant was almost identical to wild type when grown under high CO2 conditions as well as after shifts to low CO2 conditions. However, measurements of the photosystem I redox kinetic in cells of the Δsml0013::Km mutant revealed differences to wild type such as a decreased capability of cyclic electron flow as well as of utilization of electrons from catabolic processes. These results suggest that the Sml0013 protein (named NdhP) represent a novel subunit of the cyanobacterial NDH1 complex mediating its coupling to the respiratory or photosynthetic electron flow.