Two-step polyhydroxybutyrate production from hydrogenic effluent by freshwater microalgae sp. KKU-P1 and sp. KKU-P2 under mixotrophic cultivation.

This study aimed to produce PHB using hydrogenic effluent discharged from the biohydrogen production process with freshwater microalgae including sp. KKU-P1, and sp. KKU-P2. Batch experiments explored the influence of initial pH and hydrogenic effluent concentration, revealing optimal conditions at 10 % (v/v) effluent concentration and a pH of 6.5 for both KKU-P1 and KKU-P2. Subsequently, medium formulation and photoperiods were optimized to maximize biomass and PHB accumulation. The results showed that the optimal condition for PHB accumulation with KKU-P1 and KKU-P2 was nitrogen phosphorus (NP)-limited Bold's Basal Medium (BBM) under dark conditions. A two-step PHB accumulation in the upscale bioreactor was investigated under optimal conditions. The results showed that KKU-P1 achieved maximum PHB, protein, carbohydrate, and lipid contents of 4.57 %, 29.37 %, 24.76 %, and 13.21 %, respectively, whereas KKU-P2 achieved 6.35 %, 31.53 %, 16.16 %, and 4.77 %, respectively. Based on these findings, it appears that a mixotrophic approach under nutrient-limiting conditions is effective for PHB production in both KKU-P1 and KKU-P2 strains.