This study aimed to evaluate energy expenditure (EE) and heart rate (HR) response in kart pilots to successive driving bouts during a 24-hour team race. Eight adult male pilots (22.8 ± 4.1 years) participated to a team 24-hour speedway kart race in Le Mans (France). They alternatively piloted a 390 cm kart. Each relay was 45 minutes long and each pilot performed 4 relays. For each pilot, mean speeds were calculated from lap-to-lap duration recordings using a telemetric infrared timing device. Heart rate values were recorded continuously on 5-second intervals using a portable cardiometric device. Total energy expenditure (EET) and physical activity ratio (PAR) were determined by accelerometry. To pilot a kart during 45 minutes at a mean speed around 62 km·h induces a 300-kcal EET, corresponding to a 5.6-Mets PAR. This effort is responsive for a 73 b·min increase in HR, from 84.1 ± 7.6 to 157.4 ± 11.0 b·min (82% maximal heart rate intensity). However, during this relay period, HR values seemed independent to mean speed performance and bioenergetical values. Thus, in the context of the 24-hour team race, the variability in effort made during each relay and relay succession did not alter bioenergetical adaptation of pilots to kart driving. The high EE and HR values would be better explained by both emotional stress and environmental constraints such as speedway configuration and vibrations. The way how these factors specifically influence bioenergetical demand, and their relative importance, has to be specified to optimize training procedure and recommendations.