The 2022 Kunming-Montreal Global Biodiversity Framework calls upon the private sector to take substantial action to mitigate its negative impacts on biodiversity and contribute towards nature recovery. The term ‘Nature Positive’ has gained traction in biodiversity conservation discourse to describe both a societal goal and the ambitions of individual organisations to halt and reverse nature loss. However, enabling businesses to contribute towards Nature Positive outcomes will require major shifts in the way businesses and society operate, and research that can help guide and prioritise business actions. As a group of researchers and consultants working at the interface between business and biodiversity, we propose a conceptual model through which private sector contributions to a Nature Positive future could be realised and use it to identify priority research questions. The key questions address: i) sectoral strategic options, ii) methods and approaches individual businesses can implement to inform these strategies, iii) systemic driving forces that influence private sector action, and iv) how outcomes are measured to deliver Nature Positive contributions.Collaborations between researchers, businesses and industry bodies are needed to co-design and implement research, where there is currently no coordinated approach to identify and fund priority research areas for Nature Positive themes. A clearly structured and prioritised research agenda is vital to guide effective, equitable and timely action by businesses.