Temperature sensitivity (Q₁₀) of soil organic matter (SOM) decomposition is an important parameter in models of the global carbon (C) cycle. Previous studies have suggested that substrate quality controls the intrinsic Q₁₀, whereas environmental factors can impose large constraints. For example, physical protection of SOM and its association with minerals attenuate the apparent Q₁₀ through reducing substrate availability and accessibility ([S]). The magnitude of this dampening effect, however, has never been quantified. We simulated theoretical Q₁₀ changes across a wide range of [S] and found that the relationship between Q₁₀ and the log₁₀-transformed [S] followed a logistic rather than a linear function. Based on the unique Holocene paleosol chronosequence (7 soils from ca. 500 to 6900 years old), we demonstrated that the Q₁₀ decreased nonlinearly with soil age up to 1150 years, beyond which Q₁₀ remained stable. Hierarchical partitioning analysis indicated that an integrated C availability index, derived from principal component analysis of DOC content and parameters reflecting physical protection and mineral association, was the main explanatory variable for the nonlinear decrease of Q₁₀ with soil age. Microbial inoculation and ¹³C-labelled glucose addition showed that low C availability induced by physical protection and minerals association attenuated Q₁₀ along the chronosequence. A separate soil incubation experiment indicated that Q₁₀ increased exponentially with activation energy (E_a) in the modern soil, suggesting that SOM chemical complexity regulates Q₁₀ only when C availability is high. In conclusion, organic matter availability strongly decreased with soil age, whereas Michelis–Menten kinetics defines the Q₁₀ response depending on C availability, but Arrhenius equation describes the effects of increasing substrate complexity.