PurposeThis work aims to analyze the university–industry interaction in innovation ecosystems. The problem under study addresses how agents can operate in university–industry interactions to enhance the connections, specifically in the aspect of operational mechanisms of an innovation ecosystem.Design/methodology/approachThe study is qualitative, applied through grounded theory approach, cross-sectional and multiple sources of evidence – semi-structured interviews were conducted. An explanatory conceptual model was subsequently compared with the literature and validated with specialists.FindingsThe results showed that the university–industry interaction is influenced by several factors, such as networking, legal support, facilitating agents, management practices. Despite this, some other factors were identified for the improvement of U–I relationships, such as: strengthening of triple helix, greater legal certainty and encouragement of open innovation.Research limitations/implicationsThe small sample size and the heterogeneity among the universities interviewed did not allow full saturation to occur. In spite of that, a significant level of saturation with respect to the challenges and barriers was observed.Practical implicationsThis work has a direct dialogue with researchers, managers and actors involved in university–industry interaction with regard to increasing the capacity for creation and dissemination of knowledge among organizations, educational institutions, government and companies.Originality/valueBy adopting a grounded theory approach, a detailed research agenda addresses research needs in two main areas: activities that precede the interaction between universities and companies, and the organization and management of the consequences of collaborative relationships.