Algal Hydrogen Production and Exopolysaccharide Patterns in Chlorella-Bacillus Inter-Kingdom Co-Cultures

Biohydrogen production from wastewater using eukaryotic green algae can be facilitated by appropriately selected bacterial partners and cultivation conditions. Two Chlorella algal species were chosen for these experiments, based on their robust growth ability in synthetic wastewater. The applied three Bacillus bacterial partners showed active respiration and efficient biomass production in the same synthetic wastewater. Bacillus amyloliquefaciens, Bacillus mycoides, and Bacillus cereus as bacterial partners were shown to specifically promote algal biomass yield. Various inter-kingdom co-culture combinations were investigated for algal-bacterial biomass generation, for co-culture-specific exopolysaccharide patterns, and, primarily, for algal biohydrogen evolution. Chlorella sp. MACC-38 mono- and co-cultures generated significantly higher biomass compared with that of Chlorella sp. MACC-360 mono- and co-cultures, while in terms of hydrogen production, Chlorella sp. MACC-360 co-cultures clearly surpassed their Chlorella sp. MACC-38 counterparts. Imaging studies revealed tight physical interactions between the algal and bacterial partners and revealed the formation of co-culture-specific exopolysaccharides. Efficient bacterial respiration was in clear correlation with algal hydrogen production. Stable and sustainable algal hydrogen production was observed in synthetic wastewater for Chlorella sp. MACC-360 green algae in co-cultures with either Bacillus amyloliquefaciens or Bacillus cereus. The highest algal hydrogen yields (30 mL H-2 L-1 d(-1)) were obtained when Chlorella sp. MACC-360 was co-cultured with Bacillus amyloliquefaciens. Further co-culture-specific algal biomolecules such as co-cultivation-specific exopolysaccharides increase the valorization potential of algal-bacterial co-cultures and might contribute to the feasibility of algal biohydrogen production technologies.