Bioaugmentation with Clostridium pasteurianum for high-yield continuous bio-hydrogen production in a dynamic membrane bioreactor

Abstract

This study demonstrated the feasibility of achieving high-yield continuous bio-H2 production through bioaugmentation with Clostridium pasteurianum. C. pasteurianum was introduced during the initial granulation stage in a dynamic membrane bioreactor (DMBR) inoculated with heat-treated digester sludge. Following bioaugmentation, the average hydrogen yield (HY) reached 3.07 ± 0.13 mol H2/mol hexoseconsumed, with a hydraulic retention time (HRT) of 2 h. This high-yield and high-rate hydrogen production performance was achieved alongside a high concentration of suspended and granular biomass as 21.02 ± 1.01 g VSS/L. The DM played a crucial role in prolonging the solid retention time of the suspended biomass to 12.15 h. C. pasteurianum emerged as the predominant species, constituting 92.3 % of the total bacterial population. This dominance significantly contributed to achieving the high HY, reaching 77 % of the theoretical maximum HY (4 mol H2/mol hexoseconsumed) by shifting the metabolic pathway from non-H2 production pathways to hydrogen-producing acetate. However, during further continuous operation, the hydrogen production pathway was gradually shifted towards the butyrate pathway, resulting in a decreased HY of 2.26 mol H2/mol hexoseconsumed along with the decreased population of C. pasteurianum. Additional bioaugmentation of C. pasteurianum during the maturational granulation stage did not impact the metabolic pathways or microbiome significantly, as the added strain was supposed to be washed out. This study provides insights into achieving high-yield continuous bio-H2 production through bioaugmentation with C. pasteurianum during the initial granulation stage. Further studies are necessary to sustain C. pasteurianum-dominated high HY with the acetate pathway during prolonged operation.