Purpose <p>Understanding soil enzyme activity dynamics and stoichiometry is essential for assessing ecosystem nutrient use efficiency and addressing microbial metabolic limitations. The interactive effects of warming and increasing copper pollution in agricultural soils are of growing concern. However, the impact of copper on soil enzyme activities and stoichiometry under warming conditions remains unclear. Consequently, investigating soil enzyme stoichiometric responses to copper gradients under simulated warming is crucial to elucidate soil microbial nutrient balance.</p> Materials and methods <p>This study used a split-plot experimental design with temperature as the main plot treatment and copper concentration as the subplot treatment. The temperature treatments (T) included nighttime warming (HT) and no warming (NT). The copper treatments included control (CK), low (LP), medium (MP), and high (HP) concentrations. The dynamics of soil enzyme activities [catalase (CAT), Alkaline phosphatase (ALP), and urease (UE)], enzyme stoichiometry [EEA_{C: N} (enzyme C: N ratio), EEA_{C: P} (enzyme C: P ratio), and EEA_{N: P} (enzyme N: P ratio)], and microbial nutrient limitations were analysed using a long-term in situ passive warming platform (&gt; 10 years operational).</p> Results and discussion <p>The results revealed that warming significantly reduced CAT activity (42.5–51.4%) and microbial C limitations (27.9–37.9%) across all copper treatments at jointing stage. Under NT, UE and ALP activities significantly decreased in MP and HP at jointing and filling stages. All copper treatments significantly elevated EEA_{C: P} (17.0-39.6%) and microbial C limitations (20.2–31.2%) at heading stage. Warming significantly reduced UE activity by 17.3% in MP at maturity stage, but significantly increased ALP activity by 55.3% in HP at heading stage. At filling stage, warming significantly reduced microbial C limitations by 44.3% and 50.0% in LP and HP, respectively. Soil moisture, copper, aboveground biomass, pH and available P were the main factors influencing enzyme activity and stoichiometric characteristics.</p> Conclusions <p>Copper inhibition on soil enzyme activities and elevation in enzyme stoichiometric ratios demonstrated growth-stage dependence. Under NT, low and medium copper treatments increased microbial C and P limitations at different growth stages. Warming altered responses of enzyme activities, stoichiometric ratios, and microbial nutrient limitations to copper. It alleviated microbial C limitations under low and high copper treatments, but intensified microbial P limitations under high copper treatment.</p>