The purpose of this study is to determine the relationship between ability, motivation, and opportunity as expressed in human resource management (HRM) practices and corporate entrepreneurship (CE) in Yemeni public universities. It also uses organizational culture (OC) as a moderator variable to find out more about the indirect relationship between HRM practices and CE. HRM practices that organize human and social capital are critical for enhancing and facilitating CE. Nevertheless, there are variations in the published results about the impact of OC (as a moderator) on the relationship between ability, motivation, and opportunity with CE. The validity of HRM practices and the CE model will be tested empirically using data obtained from academic staff at Yemeni universities, with a sample size of 203 academic staff. SmartPLS 3.0 software was used, and the framework is made up of ability, motivation, opportunity, OC, and CE. The findings from the direct relationships demonstrate the significance of ability, motivation, and opportunity elements in fostering CE, particularly in Yemeni public universities. Furthermore, the moderate impact of organizational culture on the proposed relationship was investigated. Therefore, organizational culture (OC) moderates the association between ability and CE in a positive manner. In contrast, OC does not influence the relationship between motivation and CE. Additionally, OC has no moderating impact on the relationship between opportunity and CE. Moreover, the findings of study will help university leaders, scholars, policymakers, and people in charge of making decisions in the higher education sector make policies to improve HRM practices and CE. In general, HRM practices offer a new perspective on CE by utilizing an ability–motivation–opportunity (AMO). Thus, these HRM factors can be used as a guide to show how the best use of corporate resources can be made at different stages of the CE process.