Abstract Energy uncoupling technology was applied to the membrane process to control the problem of bio-fouling. Different dosages of uncoupler (2,4‑dinitrophenol, DNP) were added to the activated sludge, and a short-term ultrafiltration test was systematically investigated for analyzing membrane fouling potential and underlying mechanisms. Ultrafiltration membrane was used and made of polyether-sulfone with a molecular weight cut off (MWCO) of 150 kDa. Results indicated that low DNP concentration (15–30 mg/g VSS) aggravated membrane fouling because more soluble microbial products were released and then rejected by the membrane, which significantly increased cake layer resistance compared with the control. Conversely, a high dosage of DNP (45 mg/g VSS) retarded membrane fouling owing to the high inhibition of extracellular polymeric substances (proteins and polysaccharides) of the sludge, which effectively prevented the formation of cake layer on the membrane surface. Furthermore, analyses of fouling model revealed that a high dosage of DNP delayed the fouling model from pore blocking transition to cake filtration, whereas this transition process was accelerated in the low dosage scenario. Graphical abstract Unlabelled Image Highlights • Effects of DNP addition to the sludge on membrane bio-fouling were evaluated. • Low dosage of DNP significantly accelerated the cake layer resistance of fouling. • Because more SMPs were released and rejected by membrane forming denser layer. • High DNP retarded membrane fouling due to high inhibition of sludge EPS. • High DNP delayed the transition of fouling model from pore blocking to cake layer. [ABSTRACT FROM AUTHOR]