Background: Music therapy is a promising complementary intervention for addressing various mental health conditions. Despite evidence of the beneficial effects of music, the acoustic features that make music effective in therapeutic contexts remain elusive. Aims: This study aimed to identify and validate distinctive acoustic features of healing music. Methods: We constructed a healing music dataset (HMD) based on nominations from related professionals and extracted 370 acoustic features. Healing-distinctive acoustic features were identified as those that were (1) independent from genre within the HMD, (2) significantly different from music pieces in a classical music dataset (CMD) and (3) similar to pieces in a five-element music dataset (FEMD). We validated the identified features by comparing jazz pieces in the HMD with a jazz music dataset (JMD). We also examined the emotional properties of the features in a Chinese affective music system (CAMS). Results: The HMD comprised 165 pieces. Among all the acoustic features, 74.59% shared commonalities across genres, and 26.22% significantly differed between the HMD classical pieces and the CMD. The equivalence test showed that the HMD and FEMD did not differ significantly in 9.46% of the features. The potential healing-distinctive acoustic features were identified as the standard deviation of the roughness, mean and period entropy of the third coefficient of the mel-frequency cepstral coefficients. In a three-dimensional space defined by these features, HMD's jazz pieces could be distinguished from those of the JMD. These three features could significantly predict both subjective valence and arousal ratings in the CAMS. Conclusions: The distinctive acoustic features of healing music that have been identified and validated in this study have implications for the development of artificial intelligence models for identifying therapeutic music, particularly in contexts where access to professional expertise may be limited. This study contributes to the growing body of research exploring the potential of digital technologies for healthcare interventions.