Motivation is a fundamental element for the practice of physical activity and the feeling of satisfaction with life. However, little is known about the role of such psychological variables in the lifestyle of Chilean university students. This study aimed to analyze the relationship between life satisfaction and the regulations of the motivation, and to compare such variables by physical activity level and sedentary behavior of university students. The sample comprised 95 Chilean university students (63.2% female), with a mean age of 20.92 ± 1.98 years. The instruments were the IPAQ, the Satisfaction with Life Scale, and the Behavioral Regulation in Exercise Questionnaire. Descriptive measures, Pearson’s correlation coefficient, and Student’s t tests were used to analyze the data. The results revealed positive and significant correlations between life satisfaction and intrinsic (r = 0,44), integrated (r = 0,38), and identified (r = 0,41) regulation. Students with high levels of physical activity scored higher for intrinsic, integrated, and identified regulation. Those who reported low sedentary behavior showed higher identified regulation. Therefore, it is suggested that students’ motivational regulation may differ on some dimensions as those with higher levels of physical activity show more adaptive motivational regulation. Identified regulation also seems to play an important role in decreasing sedentary behavior.