Extracellular polymeric substance decomposition linked to hydrogen recovery from waste activated sludge: Role of peracetic acid and free nitrous acid co-pretreatment in a prefermentation-bioelectrolysis cascading system

Free nitrous acid (FNA) has been recently reported to be an effective and eco-friendly inactivator for waste activated sludge (WAS), while the limited decomposition of the extracellular polymeric substance (EPS) matrix hampers resource recovery from WAS. This work employed peracetic acid (PAA) to assist FNA and explored the contribution of co-pretreatment to hydrogen recovery in a prefermentation-bioelectrolysis cascading system. The results showed that co-pretreatment led to approximately 8.8% and 20.4% increases in the exfoliation of particulate proteins and carbohydrates, respectively, from tightly bound EPS (TB-EPS) over that of sole FNA pretreatment. Electron paramagnetic resonance analysis verified that the synergistic effect of FNA, PAA and various generated free radicals was the essential process. This effect further promoted the accumulation of volatile fatty acids (VFAs) after 96 h of prefermentation, and the peak concentration in co-pretreated WAS (AD-FPWAS) was approximately 2.5-fold that in sole FNA-pretreated WAS (AD-FWAS). Subsequently, the cascading utilization of organics in the bioelectrolysis step contributed to efficient hydrogen generation. A total of 10.8 ± 0.3 mg H₂/g VSS was harvested in microbial electrolysis cells (MECs) fed with AD-FPWAS, while 6.2 ± 0.1 mg H₂/g VSS was obtained from AD-FWAS. X-ray photoelectron spectroscopy (XPS) revealed the effective decomposition of the phospholipid bilayer in the cytomembrane and the transformation of macromolecular organics into VFAs and hydrogen in the cascading system. Further microbial community analysis demonstrated that co-pretreatment enhanced the accumulation of functional consortia, including anaerobic fermentative bacteria (AFB, 28.1%), e.g., Macellibacteroides (6.3%) and Sedimentibacter (6.9%), and electrochemically active bacteria (EAB, 57.0%), e.g., Geobacter (39.0%) and Pseudomonas (13.6%), in the prefermentation and MEC steps, respectively. The possible synergetic and competitive relationships among AFB, EAB, homo-acetogens, nitrate-reducing bacteria and methanogens were explored by molecular ecological network analysis. From an environmental and economic perspective, this promising FNA and PAA co-pretreatment approach provides new insight for energy recovery from WAS biorefineries.