Volatile fatty acid (VFA) production stimulated by saponin (SP), an environmentally friendly bio-surfactant, was investigated during sludge alkaline fermentation in laboratory studies and pilot applications. The combined use of SP and pH 9 condition significantly enhanced VFA production to approximately 425 mg COD/g VSS, which was 4.7-fold of raw sludge and 1.5-fold of sole pH 10 adjustment (the optimum pH for alkaline fermentation). Further results indicated that SP & pH 9 condition provided sufficient substrates for acidification and decreased the consumption of VFAs through methanogenesis. Moreover, SP accompanied by moderate alkaline condition (i.e. pH 9) showed weaker inhibitory effects on key enzyme activities and metabolic potential of acidification microorganisms than sole pH 10 adjustment. On this basis, a pilot-scale system involving anaerobic fermentation and anaerobic-anoxic-aerobic step-feed bioreaction tanks was established to study the potential of VFAs as supplementary carbon sources for wastewater treatment. The influent of the pilot system was sanitary wastewater characterized by low C/N ratios from a scenic rural area. After flocculation and nutrient precipitation, the fermentation supernatant was mixed with the influent at a volume ratio of 1:30. With this approach, nitrogen and phosphorus concentrations in effluent fulfilled the first-A wastewater discharge standard in China.