Mobility as a Service (MaaS) is a promising concept which aims at offering seamless mobility to end users and providing economic, societal, transport-related and environmental benefits to the cities of the future. To achieve a successful future market take-up of MaaS it is important to develop prototype business models to offer high-value bundled mobility services to customers, as well as enable the MaaS operator and the involved actors to capture value. This paper aims at investigating the business perspective of MaaS by collecting qualitative data from workshops and in-depth interviews in three European metropolitan areas: Budapest, Greater Manchester and the city of Luxembourg. The analysis of the collected data contributed to the in-depth analysis of the MaaS business ecosystem and the identification of the champions of MaaS in the three areas. Prototype business models for MaaS are developed based on the Osterwalder’s canvas, to describe how MaaS operators may create, deliver, and capture value. Our findings indicate that the MaaS ecosystem comprises of public and private actors who need to cooperate and compete in order to capture value. Although noticeable deviations among the study areas are observed, mobility service providers, public transport authorities and regional authorities were commonly indicated as the key actors in a MaaS partnership. In addition, viewed as a system, enablers and barriers to MaaS are identified based on the systems’ of innovation approach. The analysis indicates that the regulatory framework of the cities, the lack of standardization and openness of the application programming interfaces and the need for transport-related investments constitute risks for the successful implementation of MaaS in the study areas. Trust between MaaS actors and cooperation in e-ticketing are key enablers in some of the study areas.