Chronic nitrogen (N) and (or) sulfur (S) deposition to boreal forests in the Athabasca oil sands region (AOSR) in Alberta, Canada, has been caused by oil sands mining and extraction/upgrading activities. It is important that we understand the response of microbial community function to chronic N and S deposition as microbial populations mediate soil carbon (C) and N cycles and affect ecosystem resilience. To evaluate the impact of N and (or) S deposition on soil microbial community functions, we conducted a simulated N and S deposition experiment in a boreal mixedwood forest with the following four treatments: control (CK), N addition (+N, 30 kg N·ha⁻¹ as NH₄NO₃), S addition (+S, 30 kg S·ha⁻¹ as NaSO₄), and N plus S addition (+NS, 30 kg N·ha⁻¹ + 30 kg S·ha⁻¹), from 2006 to 2010. Nitrogen and (or) S deposition did not change soil organic carbon, total N, dissolved organic C and N, or soil microbial biomass C and N. Soil microbial community-level physiological profiles, however, were strongly affected by 5 years of N and (or) S addition. Soil β-glucosidase activity in the +NS treatment was greater than that in the +S treatment, and S addition decreased soil arylsulfatase; however, urease and dehydrogenase activities were not affected by the simulated N and (or) S deposition. Our data suggested that N and (or) S deposition strongly affected soil microbial community functions and enzymatic activities without changing soil microbial biomass in the studied boreal forest.