Antibiotics in waste activated sludge (WAS) has drawn increasing attention because of their persistent and bioaccumulation characteristics. Most study illustrated the role of antibiotics in anaerobic fermentation from WAS, but lacking the analysis at microbial level as well as the possible interaction between them. This study investigated the effect of three sulfonamide antibiotics (sulfamethoxazole (SMX), sulfaquinoxaline (SQX), and sulfadiazine (SD)) on WAS fermentation and explored its microbiological mechanism. Results indicated that the production of short-chain fatty acids (SCFAs) was significantly improved by 1.9 folds with a peak value at 4626.1 mg COD L−1 in the existence of SD. This was attributed to the promoted release of soluble proteins and polysaccharides with the existence of sulfonamide antibiotics (SAs) as revealed by the excitation-emission matrix (EEM) spectrum. Analysis of microbial community structure showed that the total abundance of the fermenters in groups with SAs was1.2–1.6 times of that in Control. Specifically, the acid-forming genus Tissierella in SMX and SQX increased by 12.1%–15.0% compared with the Control, while the proteolytic genus Proteinivorax dominated in SD with 39.5%. Molecular ecological networks (MENs) analysis further revealed the potential cooperative relationships among different fermenters. This study was anticipated to provide some valuable information for the behavior of antibiotics in WAS fermentation. Display omitted •The existence of SMX, SQX and SD facilitated SCFAs production from WAS anaerobic fermentation.•The presence of SMX, SQX and SD enhanced the solubility and biodegradability of proteins and polysaccharides.•The abundance of fermenters was increased in fermentation systems containing SMX, SQX and SD.•The potential cooperative relationships were established among the key microbial community.