Agribusiness is central to human life, and eco-innovation is the key driving force behind economic and ecological growth. However, in developing countries, setting economic and environmental goals remains challenging for entrepreneurs. This article focuses on the perceived role of entrepreneurial opportunity identification (EOI) factors on eco-innovation in Tunisian agribusiness. A combination of external and sustainable factors was used to create an integrated model to assess the potential of EOI for stimulating eco-innovation and sustainable development. Structural equation modelling was used to explore the positive effect of these factors and investigate the mediating role of entrepreneurial alertness. The findings confirm (a) the positive influence of EOI factors on eco-innovation, which serves as an innovative way for entrepreneurs to gain profits; (b) entrepreneurial alertness constitutes the most important factor for adopting eco-innovation; (c) entrepreneurial alertness plays a mediating role between (1) prior knowledge and (2) social networks and eco-innovation.