The recently developed Denitrifying Sulfur conversion-associated Enhanced Biological Phosphorus Removal (DS-EBPR) process has demonstrated simultaneous removal of organics, nitrogen and phosphorus with minimal sludge production in the treatment of saline/brackish wastewater. Its performance, however, is sensitive to operating and environmental conditions. In this study, the effects of temperature (20, 25, 30 and 35 degrees C) and the ratio of influent acetate to propionate (100-0, 75-25, 50-50, 25-75 and 0 - 100%) on anaerobic metabolism were investigated, and their optimal values/controls for performance optimization were identified. A mature DS-EBPR sludge enriched with approximately 30% sulfate reducing bacteria (SRB) and 33% sulfide-oxidizing bacteria (SOB) was used in this study. The anaerobic stoichiometry of this process was insensitive to temperature or changes in the carbon source. However, an increase in temperature from 20 to 35 degrees C accelerated the kinetic reactions of the functional bacteria (i.e. SRB and SOB) and raised the energy requirement for their anaerobic maintenance, while a moderate temperature (25-30 degrees C) resulted in better P removal (>= 93%,18.6 mg P/L removal from total 20 mg P/L in the influent) with a maximum sulfur conversion of approximately 16 mg S/L. These results indicate that the functional bacteria are likely to be mesophilic. When a mixed carbon source (75-25 and 50-50% acetate to propionate ratios) was supplied, DS-EBPR achieved a stable P removal (>= 89%, 17.8 mg P/L for 400 mg COD/L in the influent) with sulfur conversions at around 23 mg S/L, suggesting the functional bacteria could effectively adapt to changes in acetate or propionate as the carbon source. The optimal temperatures or carbon source conditions maximized the functional bacteria competition against glycogen-accumulating organisms by favoring their activity and synergy. Therefore, the DS-EBPR process can be optimized by setting the temperature in the appropriate range (25-30 degrees C) and/or manipulating influent carbon sources.