Purpose: This study aims to take a whole-of-chain perspective to explore how entrepreneurial orientation (EO) influences collaborative performance in agri-food value-chains through enhancing the acquisition of knowledge. Design/methodology/approach: Data were collected through a survey of 233 actors, including farmers, intermediaries and retailers in one beef cattle value-chain in the Central Highlands, Vietnam. Structural equation modeling was used to test hypotheses. Findings: The actors’ level of EO within a value-chain is positively associated with collaborative performance within the beef value-chain. Additionally, knowledge acquisition partially mediates the relationship between EO and collaborative performance. Research limitations/implications: Generalizability is limited because of sampling constraints. Originality/value: To the best of the authors’ knowledge, this is the first study of EO from a whole-of-chain perspective in agri-food value-chains in an emerging economy and has implications for policymakers and agri-food marketers.