Development of oriented multi-enzyme strengthens waste activated sludge disintegration and anaerobic digestion: Performance, components transformation and microbial communities

Low methane production and long retention time are the main dilemmas in current anaerobic digestion (AD) of waste activated sludge (WAS). This work used WAS as only substrate to prepare oriented multi-enzyme (ME) that directly used for WAS pretreatment. Under the optimal parameters, the highest activities of protease and amylase in ME could respectively reach 16.5 U/g and 580 U/g, and the corresponding methane production attained 197 mLCH₄/g VS, which was increased by 70.4% compared to blank group. It was found that ME pretreatment could strengthen WAS disintegration and organic matters dissolution, lead to the soluble chemical oxygen demand (SCOD) was increased from the initial 486 mg/L to 2583 mg/L, and the corresponding volatile suspended solid (VSS) and extracellular polymeric substances (EPS) were reduced by 27% and 73.8%, respectively. The results of three-dimensional excitation-emission matrix (3D-EEM) and Fourier transform infrared spectroscopy (FTIR) indicated that protein disintegration may be the critical step during the process of WAS hydrolysis with ME, of which the release of tyrosine-like proteins achieved the better biodegradability of WAS, while the results of X-ray photoelectron spectroscopy (XPS) showed that the formation of protein derivatives was the main harmful factor that could extend the lag phase of AD process. Microbial communities analysis further suggested that ME pretreatment facilitated the enrichment of acetogenic bacteria and acetotrophic methanogens, which caused the transition of the methanogenesis pathway from hydrogenotrophic to acetotrophic. This study is expected to furnish valuable insight for ME pretreatment on enhancing WAS disintegration and methane production.