Background: This study intends to investigate the relationship between organizational ambidexterity and knowledge management practices, with a focus on travel and tourism industries specifically. The research is conducted using a descriptive-correlation approach and involves 72 businesses located in Qom, Mashhad, and Tehran. A total of 369 managers and personnel from these organizations participated in the study, providing valuable insights into the dynamics of knowledge management and its impact on organizational learning and ambidexterity within the tourism sector. Methods: In this study, descriptive-correlation research methodology was used. Data were collected through a questionnaire distributed to 369 managers and personnel from 72 tourism organizations. The questionnaire focused on assessing the participants' perspectives on knowledge management strategies, organizational learning capability, and organizational ambidexterity. This quantitative approach allowed for the examination of relationships and mediation effects within the specified variables. Findings: The results of the study show substantial correlation between organizational ambidexterity, learning ability, and knowledge management techniques. It was found to be fascinating that the relationship between organizational ambidexterity and knowledge management strategies in the tourism setting was partially mediated by organizational learning capacity. Conclusion: In conclusion, tourism companies should place a high priority on implementing both explicit and tacit knowledge management strategies in order to attain structural ambidexterity. Effective HRM procedures as well as the advancement of knowledge management and information technology infrastructures must serve as the cornerstones of these strategies. Encouraging learning, unlearning, relearning, experimentation, and knowledge sharing are pivotal for enhancing organizational learning capability and, consequently, achieving organizational ambidexterity. This study underscores the need for a holistic approach that integrates human and technological elements to foster a dynamic and adaptable organizational structure in the tourism sector.