The adaptation of pulmonary O2 uptake (V̇o2p) kinetics is slowed in older compared with young adults during the transition to moderate-intensity exercise. In this study, we examined the relationship between V̇o2p kinetics and mitochondrial pyruvate dehydrogenase (PDH) activity in young ( n = 7) and older ( n = 6) adults. Subjects performed cycle exercise to a work rate corresponding to ∼90% of estimated lactate threshold. Phase 2 V̇o2p kinetics were slower ( P < 0.05) in older (τ = 40 ± 17 s) compared with young (τ = 21 ± 6 s) adults. Relative phosphocreatine (PCr) breakdown was greater ( P < 0.05) at 30 s in older compared with young adults. Absolute PCr breakdown at 6 min was greater ( P < 0.05) in older compared with young adults. In young adults, PDH activity increased ( P < 0.05) from baseline to 30 s, with no further change observed at 6 min. In older adults, PDH activity during baseline exercise was similar to that seen in young adults. During the exercise transition, PDH activity did not increase ( P > 0.05) at 30 s of exercise but was elevated ( P < 0.05) after 6 min. The change in deoxyhemoglobin (HHb) was greater for a given V̇o2p in older adults, and there was a similar time course of HHb accompanying the slower V̇o2p kinetics in the older adults, suggesting a slower adaptation of bulk O2 delivery in older adults. In conclusion, the slower adaptation of V̇o2p in older adults is likely a result of both an increased metabolic inertia and lower O2 availability.