New strategy for bioplastic and exopolysaccharides production: Enrichment of field microbiomes with cyanobacteria.

Seven photosynthethic microbiomes were collected from field environmental samples to test their potential in polyhydroxybutyrate (PHB) and exopolysaccharides (EPS) production, both alternatives to chemical-based polymers. Microscope observations together with microbial sequence analysis revealed the microbiome enrichment in cyanobacteria after culture growth under phosphorus limitation. PHB and EPS production were studied under three culture factors (phototrophy, mixotrophy and heterotrophy) by evaluating and optimizing the effect of three parameters (organic and inorganic carbon and days under light:dark cycles) by Box-Behnken design. Results showed that optimal conditions for both biopolymers synthesis were microbiome-dependent; however, the addition of organic carbon boosted PHB production in all the tested microbiomes, producing up to 14 %dcw PHB with the addition of 1.2 g acetate·L-1 and seven days under light:dark photoperiods. The highest EPS production was 59 mg·L-1 with the addition of 1.2 g acetate·L-1 and four days under light:dark photoperiods. The methodology used is suitable for enriching microbiomes in cyanobacteria, and for testing the best conditions for bioproduct synthesis for further scale up.

Process conditions affect properties and outcomes of polyhydroxyalkanoate accumulation in municipal activated sludge.

The developments of mixed culture polyhydroxyalkanoate production has been directed to maximize the biomass PHA content with limited attention to polymer quality. Direct comparison of PHA accumulation literature is challenging, and even regularly contradicting in reported results, due to underlying differences that are not well expressed. A study was undertaken to systematically compare the commonly reported process conditions for PHA accumulation by full-scale municipal activated sludge. A biomass acclimation step combined with a pulse-wise feeding strategy resulted in maximum average PHA contents and product yields. pH control and active nitrification did not result in observable effects on the PHA productivity. Under these conditions a high molecular weight polymer (1536 ± 221 kDa) can be produced. Polymer extraction recoveries were influenced by the PHA molecular weight. A standard protocol for an activated sludge PHA accumulation test including downstream processing and standardized extraction has been developed and is available as supplementary material.

Process optimization of the polyhydroxybutyrate production in the cyanobacteria Synechocystis sp. and Synechococcus sp.

The effect of four parameters (acetate, NaCl, inorganic carbon and days in darkness) affecting the polyhydroxybutyrate (PHB) production were tested and optimized for Synechococcus sp. and Synechocystis sp. using a Box-Behnken design. The optimal conditions for Synechocystis sp. were found to be 1.2 g L-1 of acetate, 4 gC L-1 of NaHCO3, 18 g L-1 of NaCl and 0 days in darkness. For Synechococcus sp., equal acetate concentration and days in darkness, and lower inorganic carbon and NaCl concentrations than those for Synechocystis sp. were needed (0.05 g L-1 inorganic carbon and 9 g L-1 NaCl). Optimal conditions were scaled up to 3 L photobioreactors. Using Synechocystis sp., 5.6 %dcw of PHB was obtained whether adding or not acetate. In opposition, a maximum of 26.1 %dcw by using acetate was reached with Synechococcus sp. These results provide an easy method to optimize the cultivation conditions to enhance PHB production with cyanobacteria.