AimTeaching is known as a high-stress occupation. Managing fatigue and promoting work engagement, teachers’ expectations and perceptions can impact work-related stress–recovery–balance and physical activity levels. The main objective of the current study was to investigate the relationship between intrinsic motivational factors and the autonomic nervous system, such as heart rate and heart rate variability regulation, sleep, and physical activity levels.MethodsThe research included 66 primary and high school teachers in Tampere, Finland, in 2020–2021. The Reiss Motivation Profile® (RMP) was selected as an instrument to provide a practical approach to understanding people’s intrinsic motivators and the effects of those on the Firstbeat®-measured heart rate and heart rate variability, and accelerometry. Stepwise multiple linear regression models were used to assess whether the RMP motives reflected Firstbeat®-measured outcomes.ResultsMotives such as Acceptance, Eating, Family, Physical Activity, Power, Tranquillity, and Vengeance were found to have a relationship with Firstbeat®-measured outcomes. An increase in the Physical Activity motive was related to lower stress and less light physical activity. Furthermore, an increase in the Physical Activity motive showed a relationship with better recovery and a higher amount of vigorous physical activity. However, the statistical analysis of heart rate and heart rate variability showed only low explanatory power (R2 = 0.111–0.140) for stress, recovery, and sleep. On the contrary, the explanatory power of measurements related to physical activity and sedentary behavior was higher (R2 = 0.171–0.298). In addition, the need for Acceptance as an important intrinsic motive that may prevent people from vigorous physical activity deserves further research.ConclusionThe results indicated that there are several factors affecting the autonomous nervous system but also behavior. The basic desires explained sedentary and physical activity behavior better than the functioning of the autonomic nervous system. The 16 life motives can serve as a valuable theory for understanding better how to encourage people to have healthier habits.