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Development of a Microalgae-Based Continuous Starch-to-Hydrogen Conversion Approach

Eukaryotic algae represent a highly heterogeneous group in terms of organization, lifestyle, and metabolic capabilities. Unicellular green microalgae are capable of biohydrogen production through direct and indirect photolysis as well as dark fermentation. Most algae hydrogen studies focus on axenic algal cultures, although these are difficult and expensive to maintain for continuous operation. Moreover, the complex interplays and metabolic fluxes between algae and bacteria in natural ecosystems provide a number of clear biological and technological benefits to large-scale functional algae-based systems. Two green algae species from the Chlamydomonas and Chlorella genera were used to engineer stable synthetic communities by incorporating a starch-degrading bacterium from the Bacillus genus into the inter-kingdom consortium. Continuous photoheterotrophic biohydrogen production was achieved by elaborating an appropriate algal–bacterial ratio and fine-tuning the culture conditions for the synthetic consortia. Medium with starch as only carbon source served as a simple model of cheap substrate for algal hydrogen generation. The engineered pairwise algal–bacterial associations showed increased biomass and biohydrogen yield compared to the axenic control conditions. Chlorella sp. MACC-360 produced a significantly higher amount of hydrogen when both the bacterium partner and starch were added to the media compared to the axenic algae. Continuous, elevated algal hydrogen production was achieved in media supplemented with 8 g L−1 starch as sole carbon source when carefully selected initial cell number values were used for the Chlorella sp. MACC-360–B. amlyloliquefaciens co-cultures.