Search Results (32)

Cyanobacteria are essential primary producers in aquatic ecosystems. Silver nanoclusters (Ag NCs) are emerging ultrasmall nanomaterials with broad applications; however, they inevitably enter aquatic environments. This study investigated the nanotoxicity of Ag NCs on model cyanobacterium Synechococcus elongatus PCC 7942 (Syn7942), revealing concentration-dependent growth inhibition with a 72 h half-maximal effective concentration of 1.19 mg L⁻¹. Ag NCs internalized into Syn7942 cells severely impaired the photosynthetic machinery, evidenced by reduced chlorophyll a content and suppressed photochemical reactions, leading to impaired electron transport. This photosynthetic dysfunction resulted in substantial reactive oxygen species generation and lipid peroxidation, as indicated by elevated levels of malondialdehydes. Although the activities of key antioxidant enzymes were enhanced, oxidative damage ultimately overwhelmed the cellular defense capacity. Furthermore, quantitative analysis of silver ions (Ag⁺) release coupled with comparative toxicity assays of Ag⁺ and Ag NCs confirmed that the toxicity originates from the Ag NCs themselves, rather than from the released Ag⁺. Transcriptional level analysis revealed that Ag NCs altered the expression of genes involved in photosynthesis and metal detoxification. Collectively, this study reveals a unique toxicity mechanism for Ag NCs distinct from that for Ag NPs and Ag⁺, highlighting the potential ecological risks posed by ultrasmall nanomaterials. Full article

The cyanobacterial circadian clock orchestrates a wide range of physiological processes. However, its involvement in extracellular vesicle (EV) biogenesis remains largely unexplored. In this study, we investigated the role of the master circadian output regulator RpaA in modulating EV production in Synechococcus elongatus PCC 7942. Deletion of rpaA simultaneously abolished the bioluminescence rhythmicity and rhythmic EV production, whereas overexpression of rpaA enhances the output of circadian clock signals in a dose-dependent manner and markedly increases the bioluminescence intensity and EV production while maintaining the normal circadian oscillation patterns. Transcriptional analyses indicated that rpaA positively regulates lpxD to promote membrane synthesis and differentially inhibits glgC transcription to optimize carbon allocation. These findings reveal a previously unrecognized link between circadian output signaling and EV biogenesis and suggest that rpaA overexpression is an effective strategy for improving the yield of EVs. It also provides novel insights and regulatory targets for facilitating the synthesis of other high-value metabolites in cyanobacterial cell factories, and holds certain practical significance for advancing cyanobacterial synthetic biology research and the application of nanocarrier technology. Full article

Water toxicity screening requires sensitive tools to rapidly detect environmental pollutants. While complex analytical methods accurately determine known contaminants, fast screening tests utilizing biological processes, such as photosynthesis, are increasingly being developed to evaluate the toxicity of environmental waters. We describe the isolation of the psychrotolerant Coccomyxa sp. LT4 from Scarisoara Ice Cave (Romania), representing the first report of green algae inhabiting this type of environment, and provide a preliminary assessment of its isolated thylakoids as novel biorecognition components for water toxicity screening. Photosynthetic activity and diuron sensitivity were measured amperometrically and compared with thylakoids from the reference cyanobacterium Synechococcus elongatus PCC 7942. The bioreceptor’s response to various pollutants and water salinities was also investigated. The microalgal thylakoids were more sensitive to diuron than the reference thylakoids, generated stable photocurrents across a broad salinity range and, when lyophilized with sucrose, retained their activity for over two years at −20 °C. Consequently, these thylakoids, isolated from a cold-environment microalga, provide a promising basis for developing biosensors for in situ toxicity screening in low-temperature aquatic ecosystems. Full article

Photosynthesis, the main energy source for life on Earth, confronts escalating challenges of high-light–high-temperature stress (HLHT). Our previous study identified a mutation in ATP synthase, F₁-α-C252Y, that significantly enhances the HLHT tolerance of Synechococcus elongatus PCC 7942 (Sye7942), although the underlying mechanism remains obscure. In this study, we found that this mutation led to elevated levels of the b subunit of F_o, F₁ subunits, and the ATP synthase within cells, without affecting ATP synthetic activity, indicating improved intracellular ATP synthesis activity. Additionally, the mutation altered the transcriptome of Sye7942, impacting the expression of genes involved in crucial processes, such as the electron transport chain, carbon fixation, and regulatory factors, which are crucial for cyanobacteria’s adaptation to stresses. Correspondingly, the mutant exhibited enhanced photosynthesis, accelerated growth, and increased glycogen under HLHT conditions, showing improved adaptation. The higher intracellular ATP synthesis activity, along with enhanced photosynthetic activity, suggests increased ATP production in the mutant under HLHT. Enhancing ATP production and remodeling the cellular transcriptome appear to be key strategies employed by the C252Y mutation for Sye7942 acclimating to HLHT. These findings provide valuable insights for enhancing photosynthetic efficiency and stress resilience in cyanobacteria and other photosynthetic organisms facing HLHT challenges. Full article

Maintaining favorable biological productivities in photosynthetic biomanufacturing systems, especially when the risk of contamination with competing microbes is high, remains a challenge to achieve while maintaining economic feasibility. This study presents a dielectrophoresis (DEP)-based microfluidic approach for isolating a desired strain within a co-culture. The cyanobacterium Synechocystis sp. PCC 6803 (a strain capable of producing the bioplastic precursor polyhydroxybutyrate, or PHB) was enriched from mixed cultures containing the competing cyanobacterium Synechococcus elongatus PCC 7942 (which does not naturally produce PHB). A DEP cascade electrode system was established to increase purification efficiency through sequential enrichment, which leveraged inherent differences in cell morphology and dielectric properties, to achieve the selective separation of these strains under physiological conditions. A substantial increase in the relative abundance of PHB-producing cells was assessed by optical microscopy and flow cytometry characterization, confirming more than five-fold reduction of the Synechococcus fraction in the refined cell mix. The presented electrokinetic platform offers a scalable and effective approach for selectively enhancing desired microbial components within microbial biomanufacturing systems, leading towards improved product yields. Full article

The 90 kDa Heat Shock Protein (Hsp90) is an essential and highly conserved molecular chaperone that supports the folding and maturation of a diverse array of client proteins across prokaryotic and eukaryotic organisms. In bacteria, HtpG, the Hsp90 homolog, plays a central role in stress response and protein homeostasis, particularly under high-temperature and other stress conditions. Despite extensive studies on HtpG from E. coli, the biochemical properties and functional roles of cyanobacterial HtpG remain poorly characterized. Here, we focus on HtpG from the cyanobacterium Synechococcus elongatus PCC 7942 (seHtpG), a model organism for photosynthesis and circadian rhythm research. We developed a method for the overexpression and purification of seHtpG in E. coli, achieving high purity and yield suitable for biochemical and structural studies. Biophysical and biochemical assays show that seHtpG forms dimers and hydrolyzes ATP at a rate of 1.9 ATP/min, 4-fold faster than that of E. coli HtpG. This work establishes seHtpG as a model for studying the roles of HtpG in cyanobacterial protein homeostasis, photosynthesis, and stress response, enabling further exploration of cyanobacterial Hsp90 in ecosystem dynamics and biotechnological applications. Full article

Solar photovoltaic technology has consistently been regarded as a crucial direction for the development of clean energy systems in the future. Bio-photovoltaics (BPV), an emerging solar energy utilization technology, is mainly based on the photosynthesis process of photoautotrophic organisms to convert solar energy into electrical energy and output a photocurrent via extracellular electron transfer. As the fundamental unit of the bio-photovoltaic system, the stability of photosynthetic microorganisms under fluctuating and stressful light and heat conditions is likely to have a significant influence on the efficiency of bio-photovoltaic devices. However, this aspect has often been overlooked in previous bio-photovoltaics research. This study took an important cyanobacteria chassis strain, Synechococ elongatus PCC 7942, as the model organism and explored the impact of physiological robustness optimization on its performance as a bio-photovoltaic functional unit. In this work, two types of BPV systems, namely the suspension mode and the biofilm attachment mode, were assembled to evaluate the electricity-generating activity of Synechococcus cells. Overall, the latter demonstrated a remarkable photoelectric output performance. When its light and temperature tolerance was enhanced through FoF1-ATP synthase engineering, the optimized Synechococcus strain exhibited stronger photosynthetic physiology and photoelectric output activity. Under the condition of a light intensity of 2400 μmol photons/m²/s, the maximum photocurrent output of the Synechococcus-based BPV device was increased significantly by 41% over the system based on the wild-type control strain. The results of this study provided a new perspective for the future development and optimization of bio-photovoltaics. Full article

Cyanobacterial cytochrome c6 (Cyt c6) is crucial for electron transfer between the cytochrome b6f complex and photosystem I (PSI), playing a key role in photosynthesis and enhancing adaptation to extreme environments. This study investigates the high-resolution crystal structures of Cyt c6 from Synechococcus elongatus PCC 7942 and Synechocystis PCC 6803, focusing on its dimerization mechanisms and functional implications for photosynthesis. Cyt c6 was expressed in Escherichia coli using a dual-plasmid co-expression system and characterized in both oxidized and reduced states. X-ray crystallography revealed three distinct crystal forms, with asymmetric units containing 2, 4, or 12 molecules, all of which consist of repeating dimeric structures. Structural comparisons across species indicated that dimerization predominantly occurs through hydrophobic interactions within a conserved motif around the heme crevice, despite notable variations in dimer positioning. We propose that the dimerization of Cyt c6 enhances structural stability, optimizes electron transfer kinetics, and protects the protein from oxidative damage. Furthermore, we used AlphaFold3 to predict the structure of the PSI-Cyt c6 complex, revealing specific interactions that may facilitate efficient electron transfer. These findings provide new insights into the functional role of Cyt c6 dimerization and its contribution to improving cyanobacterial photosynthetic electron transport. Full article

Under standard laboratory conditions, Synechococcus elongatus PCC 7942 lacks EcaA^Syn, a periplasmic carbonic anhydrase (CA). In this study, a S. elongatus transformant was created that expressed the homologous EcaA^Cya from Cyanothece sp. ATCC 51142. This additional external CA had no discernible effect on the adaptive responses and physiology of cells exposed to changes similar to those found in S. elongatus natural habitats, such as fluctuating CO₂ and HCO₃⁻ concentrations and ratios, oxidative or light stress, and high CO₂. The transformant had a disadvantage over wild-type cells under certain conditions (Na⁺ depletion, a reduction in CO₂). S. elongatus cells lacked their own EcaA^Syn in all experimental conditions. The results suggest the presence in S. elongatus of mechanisms that limit the appearance of EcaA^Syn in the periplasm. For the first time, we offer data on the expression pattern of CCM-associated genes during S. elongatus adaptation to CO₂ replacement with HCO₃⁻, as well as cell transfer to high CO₂ levels (up to 100%). An increase in CO₂ concentration coincides with the suppression of the NDH-1₄ system, which was previously thought to function constitutively. Full article

The conserved cyanobacterial protein PipX is part of a complex interaction network with regulators involved in essential processes that include metabolic homeostasis and ribosome assembly. Because PipX interactions depend on the relative levels of their different partners and of the effector molecules binding to them, in vivo studies are required to understand the physiological significance and contribution of environmental factors to the regulation of PipX complexes. Here, we have used the NanoBiT complementation system to analyse the regulation of complex formation in Synechococcus elongatus PCC 7942 between PipX and each of its two best-characterized partners, PII and NtcA. Our results confirm previous in vitro analyses on the regulation of PipX-PII and PipX-NtcA complexes by 2-oxoglutarate and on the regulation of PipX-PII by the ATP/ADP ratio, showing the disruption of PipX-NtcA complexes due to increased levels of ADP-bound PII in Synechococcus elongatus. The demonstration of a positive role of PII on PipX-NtcA complexes during their initial response to nitrogen starvation or the impact of a PipX point mutation on the activity of PipX-PII and PipX-NtcA reporters are further indications of the sensitivity of the system. This study reveals additional regulatory complexities in the PipX interaction network, opening a path for future research on cyanobacteria. Full article

The cyanobacterial strain Cyanobacterium sp. IPPAS B-1200 isolated from Lake Balkhash is characterized by high relative amounts of myristic (30%) and myristoleic (10%) acids. The remaining fatty acids (FAs) are represented mainly by palmitic (20%) and palmitoleic (40%) acids. We expressed the genes for lysophosphatidic acid acyltransferase (LPAAT; EC 2.3.1.51) and Δ9 fatty acid desaturase (FAD; EC 1.14.19.1) from Cyanobacterium sp. IPPAS B-1200 in Synechococcus elongatus PCC 7942, which synthesizes myristic and myristoleic acids at the level of 0.5–1% and produces mainly palmitic (~60%) and palmitoleic (35%) acids. S. elongatus cells that expressed foreign LPAAT synthesized myristic acid at 26%, but did not produce myristoleic acid, suggesting that Δ9-FAD of S. elongatus cannot desaturate FAs with chain lengths less than C16. Synechococcus cells that co-expressed LPAAT and Δ9-FAD of Cyanobacterium synthesized up to 45% palmitoleic and 9% myristoleic acid, suggesting that Δ9-FAD of Cyanobacterium is capable of desaturating saturated acyl chains of any length. Full article

Phycosphere niches host rich microbial consortia that harbor dynamic algae–bacteria interactions with fundamental significance in varied natural ecosystems. Hence, culturing the uncultured microbial majority of the phycosphere microbiota is vital for deep understanding of the intricate mechanisms governing the dynamic interactions, and also to provide novel and rich microbial resources, and to discover new natural bioactive metabolites. Synechococcus elongatus PCC 7942 is a robust model cyanobacterium widely used in environment, synthesis biology, and biotechnology research. To expand the number of novel phycosphere species that were brought into culture and to discover the natural bioactivities, we presented a new yellow-pigmented bacterium named ABI-127-1, which was recovered from the phycosphere of PCC 7942, using an optimized bacterial isolation procedure. Combined polyphasic taxonomic and phylogenomic characterization was performed to confidently identify the new isolate as a potential novel species belonging to the genus Qipengyuania. The observed bioactivity of strain ABI-127-1 with promoting potential towards the growth and CO₂ fixation efficiency of the host microalgae was measured. Additionally, the bacterial production of active bioflocculant exopolysaccharides was evaluated after culture optimization. Thus, these findings revealed the potential environmental and biotechnological implications of this new microalgae growth-promoting bacterium isolated from the phycosphere microenvironment. Full article

Cyanobacteria were among the oldest organisms to undertake oxygenic photosynthesis and have an essential impact on the atmosphere and carbon/nitrogen cycles on the planet. The thylakoid membrane of cyanobacteria represents an intricate compartment that houses a variety of multi-component (pigment–)protein complexes, assembly factors, and regulators, as well as transporters involved in photosynthetic light reactions, and respiratory electron transport. How these protein components are incorporated into membranes during thylakoid formation and how individual complexes are regulated to construct the functional machinery remains elusive. Here, we carried out an in-depth statistical analysis of the thylakoid proteome data obtained during light-induced thylakoid membrane biogenesis in the model cyanobacterium Synechococcus elongatus PCC 7942. A total of 1581 proteins were experimentally quantified, among which 457 proteins demonstrated statistically significant variations in abundance at distinct thylakoid biogenesis stages. Gene Ontology and KEGG enrichment analysis revealed that predominantly photosystems, light-harvesting antennae, ABC transporters, and pathway enzymes involved in oxidative stress responses and protein folding exhibited notable alternations in abundance between high light and growth light. Moreover, through cluster analysis the 1581 proteins were categorized into six distinct clusters that have significantly different trajectories of the change in their abundance during thylakoid development. Our study provides insights into the physiological regulation for the membrane integration of protein components and functionally linked complexes during the cyanobacterial TM biogenesis process. The findings and analytical methodologies developed in this study may be valuable for studying the global responses of TM biogenesis and photosynthetic acclimation in plants and algae. Full article

Due to their unique biochemical and spectroscopic properties, both heme and phycocyanobilin are widely applied in the medical and food industries. Synechocystis sp. PCC 6803 contains both heme and phycocyanin, and is capable of synthesizing phycocyanin using heme as a precursor. The aim of this study was to uncover viable metabolic targets in the porphyrin pathway from Synechocystis sp. PCC 6803 to promote the accumulation of heme and phycocyanin in the recombinant strains of microalgae. A total of 10 genes related to heme synthesis pathway derived from Synechococcus elongatus PCC 7942 and 12 genes related to endogenous heme synthesis were individually overexpressed in strain PCC 6803. The growth rate and pigment content (heme, phycocyanin, chlorophyll a and carotenoids) of 22 recombinant algal strains were characterized. Quantitative real-time PCR technology was used to investigate the molecular mechanisms underlying the changes in physiological indicators in the recombinant algal strains. Among the 22 mutant strains, the mutant overexpressing the haemoglobin gene (glbN) of strain PCC 6803 had the highest heme content, which was 2.5 times higher than the wild type; the mutant overexpressing the gene of strain PCC 7942 (hemF) had the highest phycocyanin content, which was 4.57 times higher than the wild type. Overall, the results suggest that genes in the porphyrin pathway could significantly affect the heme and phycocyanin content in strain PCC 6803. Our study provides novel crucial targets for promoting the accumulation of heme and phycocyanin in cyanobacteria. Full article

Genome streamlining, as a natural process in the evolution of microbes, has become a common approach for generating ideal chassis cells for synthetic biology studies and industrial applications. However, systematic genome reduction remains a bottleneck in the generation of such chassis cells with cyanobacteria, due to very time-consuming genetic manipulations. Synechococcus elongatus PCC 7942, a unicellular cyanobacterium, is a candidate for systematic genome reduction, as its essential and nonessential genes have been experimentally identified. Here, we report that at least 20 of the 23 over 10 kb nonessential gene regions could be deleted and that stepwise deletions of these regions could be achieved. A septuple-deletion mutant (genome reduced by 3.8%) was generated, and the effects of genome reduction on the growth and genome-wide transcription were investigated. In the ancestral triple to sextuple mutants (b, c, d, e1), an increasingly large number of genes (up to 998) were upregulated relative to the wild type, while slightly fewer genes (831) were upregulated in the septuple mutant (f). In a different sextuple mutant (e2) derived from the quintuple mutant d, much fewer genes (232) were upregulated. Under the standard conditions in this study, the mutant e2 showed a higher growth rate than the wild type, e1 and f. Our results indicate that it is feasible to extensively reduce the genomes of cyanobacteria for generation of chassis cells and for experimental evolutionary studies. Full article