<p>This paper explores the evolution of sustainable business models in the context of biodiversity loss, focusing on 3Bee, an Italian start-up dedicated to biodiversity regeneration. The study investigates how sustainable business models respond to environmental challenges. It identifies key factors driving their transformation, offering insights into the mechanisms that enable business models to evolve and integrate sustainability at their core. Adopting a qualitative case study approach, the research draws on interviews with key informants inside the start-up, as well as secondary data sources. Applying the Business Model Dynamics lens, it analyzes how 3Bee’s business model evolved through distinct phases of innovation, validation, pivoting and scaling while adapting to market opportunities and regulatory changes. The findings highlight the pivotal role of stakeholder engagement and digital technologies in shaping each stage of development, demonstrating how these factors contribute to business scaling and impact amplification. This study makes three key contributions. First, it extends Business Model Dynamics research by applying this lens to sustainable business models, investigating overlooked stages like validation, scaling, and pivoting. Second, it identifies stakeholder engagement and digital technology adoption as critical drivers of their evolution. Third, it contributes to research on business models for biodiversity and ecosystem regeneration. For practitioners, the findings provide actionable insights on leveraging technology and stakeholder engagement to develop nature-positive and regenerative strategies that align business growth with ecological and social value creation.</p>