Abstract Background Treating latex rubber sheet wastewater often leads to the generation of a rotten-egg odor from toxic H2S. To increase the treatment efficiency and eliminate H2S, purple nonsulfur bacteria (PNSB), prepared by supplementing non-sterile rubber sheet wastewater (RAW) with fermented pineapple extract (FPE), were used to treat this wastewater under microaerobic light conditions. The following 3 independent variables: chemical oxygen demand (COD), initial pH and FPE dose were investigated using the Box–Behnken design to find optimal conditions for stimulating the growth of indigenous PNSB (PNSBsi). Results The addition of 2.0% FPE into RAW, which had a COD of 2000mgL-1 and an initial pH of 7.0, significantly decreased oxidation reduction potential (ORP) value and stimulated PNSBsi to reach a maximum of 7.8logcfumL-1 within 2d. Consequently, these PNSBsi, used as inoculants, were investigated for their ability to treat the wastewater under microaerobic light conditions. A central composite design was used to determine the optimal conditions for the wastewater treatment. These proved to be 7% PNSBsi, 0.8% FPE and 4d retention time and this combination resulted in a reduction of 91% for COD, 75% for suspended solids, 61% for total sulfide while H2S was not detected. Results of abiotic control and treatment sets indicated that H2S was produced by heterotrophic bacteria and it was then effectively deactivated by PNSBsi. Conclusions The stimulation of PNSB growth by FPE under light condition was to lower ORP, and PNSBsi proved to be effective for treating the wastewater.