Synthetic consortia is an emerging field in biotechnology, offering advantages such as division of labor between the individual members and reduced risk of contamination. Consortia combining phototrophic and heterotrophic bacteria are attractive, as phototrophs can utilize CO₂ and light energy to sustain growth. Several studies have explored such systems where sucrose is most commonly used as the provided carbon source. An alternative is acetate, a smaller two-carbon molecule produced as a by-product by many microorganisms, including the model cyanobacterium Synechocystis PCC 6803 (thereafter Synechocystis). Although acetate production during phototrophic growth is typically low, previous studies have demonstrated that metabolic engineering—specifically the introduction of phosphoketolase (PK) and overexpression of phosphotransacetylase (Pta)—enabled the development of a high-producing strain (WT_PKPa_RBS_BsPta_Δacs) capable of secreting significant levels of acetate into the medium (Roussou and Lindblad 2026). In this study, Escherichia coli and Pseudomonas taiwanensis were engineered to produce 1-butanol, an industrially relevant bulk chemical, and cultivated using acetate as the sole carbon source. These strains were then individually co-cultivated with the previously engineered Synechocystis strain, WT_PKPa_RBS_BsPta_Δacs, forming two distinct consortia that were maintained for 42 days. Growth dynamics were successfully monitored, and acetate concentrations in the consortia were lower than in a corresponding axenic Synechocystis culture. At the same time, 1-butanol production was detected in the two co-cultures, demonstrating the feasibility of coupling photosynthetically derived acetate to heterotrophic production of a value-added bulk chemical. •Photosynthetic/heterotrophic synthetic consortia for 1-butanol production. • Engineered Synechocystis PCC 6803 cells produce acetate from CO2. • Modified Escherichia coli and Pseudomonas taiwanensis cells grow on acetate and produce 1-butanol.
Stamatina Roussou, Peter Lindblad· Applied Microbiology and Bio...· 0 citations
Eukaryote-associated microbes are ubiquitous, but their essential roles in the development and ecology of their host is yet to be fully understood, partly because complex associations cannot be reconstituted and, in many instances, the genetic tools to elucidate those roles are not available. Here, we report the conjugative transfer of DNA into Nostoc azollae within three Azolla fern hosts. N. azollae is a filamentous, N2-fixing, heterocyst-forming cyanobacterium which is the predominant obligate endosymbiont of the complex microbial community associated with the floating ferns of the genus Azolla. The cyanobiont provides fixed nitrogen to its host, supporting maximum growth rates without any N-fertilizer and making Azolla symbioses both ecologically and agriculturally important. Triparental mating protocols and fluorescent reporter detection were optimized for the cyanobiont isolated from the fern, allowing the further demonstration of heterologous gene expression in N. azollae driven by several promoters, including some of a CRISPR-associated transposon (CAST) system. Azolla was then treated with a cytokinin hormone to render fern shoot apexes amenable to in planta conjugation, permitting DNA transfer to, and gene expression in two distinct developmental stages of N. azollae within Azolla. These included (i) cells of filaments from the Shoot Apical Nostoc colony, the only cyanobacterial stem-cell population vertically transmitted across fern generations, and (ii) cells from differentiated filaments in early formed Azolla leaf cavities. Our approach represents a technically groundbreaking advance for the genetic engineering of cyanobacterial endosymbioses that may be useful for other symbiotic systems, opening a way to investigate these important biological entities.
Cristina Sarasa-Buisán, E. Güngör, Enrique Flores et al.· ISME Communications· 0 citations
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