Biogas valorization via continuous polyhydroxybutyrate production by Methylocystis hirsuta in a bubble column bioreactor

Creating additional value out of biogas during waste treatment has become a priority in past years. Biogas bioconversion into valuable bioproducts such as biopolymers has emerged as a promising strategy. This work assessed the operational feasibility of a bubble column bioreactor (BCB) implemented with gas recirculation and inoculated with a polyhydroxybutyrate (PHB)-producing strain using biogas as substrate. The BCB was initially operated at empty bed residence times (EBRTs) ranging from 30 to 120 min and gas recirculation ratios (R) from 0 to 30 to assess the gas-to-liquid mass transfer and bioconversion of CH₄. Subsequently, the BCB was continuously operated at a R of 30 and an EBRT of 60 min under excess of nitrogen and nitrogen feast-famine cycles of 24 h:24 h to trigger PHB synthesis. Gas recirculation played a major role in CH₄ gas-liquid transfer, providing almost fourfold higher CH₄ elimination capacities (~41 g CH₄ m^-3 h^-1) at the highest R and EBRT of 60 min. The long-term operation under N excess conditions entailed nitrite accumulation (induced by O₂ limiting conditions) and concurrent methanotrophic activity inhibition above ~60 mg N-NO₂^- L^-1. Adjusting the N-NO₃^- supply to match microbial N demand successfully prevented nitrite accumulation. Finally, the N feast-famine 24 h:24 h strategy supported a stable CH₄ conversion with a removal efficiency of 70% along with a continuous PHB production, which yielded PHB accumulations of 14.5 ± 2.9% (mg PHB mg^-1 total suspended solids × 100). These outcomes represent the first step towards the integration of biogas biorefineries into conventional anaerobic digestion plants.