Physical fitness is a powerful indicator of health. Sleep condition plays an essential role in maintaining quality of life and is an important marker that predicts physical fitness. This study aimed to determine the relationship between sleep conditions (sleep quality, sleep duration, bedtime) and multiple physical fitness indicators (body mass index (BMI), flexibility, abdominal muscle strength and endurance, cardiopulmonary endurance) in a well-characterized population of Taiwanese adults aged 23 to 65. The applied data were obtained from the National Physical Fitness Examination Survey 2014 conducted in Taiwan. We assessed the association of the sleep conditions with physical fitness performances in Taiwanese adults by using the multivariate adaptive regression spline (MARS) method with a total of 69,559 samples. The results show that sleep duration, sleep quality, and bedtime were statistically significant influence factors on physical fitness performances with different degrees. Gender was an important factor that affects the effects of daily sleep conditions on performances of physical fitness. Sleep duration was the most important factor as it was simultaneously correlated with BMI, sit-ups, and sit-and-reach indicators in both genders. Bedtime and sleep quality were only associated with sit-ups performance in both genders.