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Background Introducing Medical Licentiates (MLs) into the Zambian health system has significantly improved the availability of health services in rural areas. Despite their importance in the delivery of health services, MLs face challenges. Limited literature exists on the barriers and enablers to MLs practice in Zambia. This study aimed to investigate these factors. Methods A qualitative case study design was used to investigate the barriers and enablers to ML practice in Zambia. Data were collected through semi-structured, in-depth interviews with 18 purposively selected MLs. Thematic analysis used a top-down approach to identify and develop emerging themes. Results The study identified several barriers to ML practice, including inadequate remuneration, lack of professional recognition, strained working relationships with medical doctors and unregulated scopes of practice that blur the distinctions between the roles of MLs and medical doctors. Medical Licentiates felt they were on an inappropriate salary scale, unsuitable for holders of Bachelor of Science degrees with no clear pathway for academic advancement. The working relationships between MLs and medical doctors were mixed, with some MLs reporting acceptable interactions, while others experienced conflict. Enablers of ML practice were linked to professional fulfilment derived from exercising their full competencies and contributing to the recovery of patients. Conclusion Our study revealed significant barriers with few enablers to ML practice in Zambia. MLs are deeply demotivated by what they perceive as an inappropriate salary scale, exacerbated by an unregulated scope of work that sees them perform similar roles to those of medical doctors. Our results indicate a pressing need for the government to ensure that MLs are placed on a salary scale that appropriately reflects their qualifications and scope of work.