Optimal control of vitamin-mediated algal–bacterial co-cultures under optogenetic regulation

Global metabolome analysis of Dunaliella tertiolecta, Phaeobacter italicus R11 Co-cultures using thermal desorption - Comprehensive two-dimensional gas chromatography - Time-of-flight mass spectrometry (TD-GC×GC-TOFMS)

Microalgae, as the most promising raw material for biodiesel, can absorb and transform N, P, and organic matter in wastewater into cell components such as oil, carbohydrate, and protein. The cultivation of microalgae in wastewater provides the possibility to realize harmless treatment and resource utilization of wastewater and reduce the cost of microalgal culture. Whether in a single microalgal culture process or wastewater system, there always be a large number of bacteria interacting with microalgae, forming a complex microecological system. Although microalgal–bacterial coculture system is feasible features for the higher biomass production and better nutrient adaptability from wastewater, the precise understanding of interaction and coexistence in water bodies is still unclear. Some major challenges are to develop sustainability in the microalgal–bacterial coculture system due to the incomplete knowledge about the mechanism of communication, algal growth promotion, and effect of microalgae on the indigenous bacteria in wastewater. In the present review, nutritional interaction and signal transduction, two main interaction models of algae and bacteria, and the effect of synergism of these two models on microalgal system are summarized. Moreover, the impact of algal–bacterial coculture on algal growth and removal of pollutants in wastewater treatment process was expounded systematically to develop highly efficient consortium systems by covering the sustainability limitations. This work would provide guidance for the establishment of efficient algal–bacterial symbiosis system in the wastewater with complex microecology and nutrient environment to improve the accumulation of microalgal biomass and reduce the cost of microalgal culture.

Interaction of Scenedesmus quadricauda and native bacteria in marine biopharmaceutical wastewater for desirable lipid production and wastewater treatment

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 H2 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.

Bacteria are integral components of cnidarian holobionts, but the nature of their interactions with algal symbionts remains unresolved. While microbial vitamin provisioning has been proposed as a key service, the extent to which this occurs in symbioses like that of Symbiodinium linucheae and its microbiome remains unclear. To address this gap, we examined the growth dynamics of algal–bacterial cocultures in nutrient-depleted conditions. Symbiodinium linucheae, the algal symbiont of Aiptasia anemones (Exaiptasia diaphana), putatively possesses the cobalamin (B12)-dependent methionine synthase gene, making it dependent on B12, an essential vitamin produced by prokaryotes. We previously isolated a bacterial B12-producer, Roseibium, a fundamental member of the Symbiodiniaceae microbiome, and sequenced its genome to confirm its metabolic potential for B12 synthesis. Here, we grew axenic cultures of S. linucheae (strain SSA01) and Roseibium in B12-limited media for 12 weeks to document growth dynamics, followed by differential gene expression analysis comparing pure culture controls (axenic SSA01 or axenic Roseibium) with coculture treatments. SSA01 exhibited limited transcriptomic differences in coculture, like enrichment of GTPase activity, ligase and genes related to replication, yet it grew at higher densities than SSA01 in monocultures, regardless of B12 availability. Roseibium also achieved higher densities in coculture, and transcriptomic differences suggest it benefits from associations with SSA01 in low nutrient environments. Taken together, these results suggest SSA01 may not be true B12 auxotrophs but may receive alternative, possibly indirect, metabolic benefits from Roseibium.

The growth dynamics of populations of interacting species in the aquatic environment is of great importance, both for understanding natural ecosystems and in efforts to cultivate these organisms for industrial purposes. Here we consider a simple two-species system wherein the bacterium Mesorhizobium loti supplies vitamin B12 (cobalamin) to the freshwater green alga Lobomonas rostrata, which requires this organic micronutrient for growth. In return, the bacterium receives photosynthate from the alga. Mathematical models are developed that describe minimally the interdependence between the two organisms, and that fit the experimental observations of the consortium. These models enable us to distinguish between different mechanisms of nutrient exchange between the organisms, and provide strong evidence that, rather than undergoing simple lysis and release of nutrients into the medium, M. loti regulates the levels of cobalamin it produces, resulting in a true mutualism with L. rostrata. Over half of all microalgae are dependent on an exogenous source of cobalamin for growth, and this vitamin is synthesised only by bacteria; it is very likely that similar symbiotic interactions underpin algal productivity more generally.

Communities of microbes play important roles in natural environments and hold great potential for deploying division-of-labor strategies in synthetic biology and bioproduction. However, the difficulty of controlling the composition of microbial consortia over time hinders their optimal use in many applications. Here, we present a fully automated, high-throughput platform that combines real-time measurements and computer-controlled optogenetic modulation of bacterial growth to implement precise and robust compositional control of a two-strain E. coli community. In addition, we develop a general framework for dynamic modeling of synthetic genetic circuits in the physiological context of E. coli and use a host-aware model to determine the optimal control parameters of our closed-loop compositional control system. Our platform succeeds in stabilizing the strain ratio of multiple parallel co-cultures at arbitrary levels and in changing these targets over time, opening the door for the implementation of dynamic compositional programs in synthetic bacterial communities.