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BioINSouth project aims to support regional policymakers in integrating ecological limits into bioeconomy strategies and roadmaps for circular bio-based activities. By developing guidelines and digital tools within the safe and sustainable by design (SSbD) assessment framework, the project fosters innovative methodologies for assessing environmental impacts across various industrial bio-based systems. Targeting Southern Mediterranean European regions, particularly those lagging behind such as Cyprus, Slovenia, Greece, and Portugal, BioINSouth will also involve broader participation from Spain, France, Italy, and international cooperation with Türkiye. Leveraging the BIOEAST initiative that focuses on central Europe, BioINSouth will establish QH-based regional Multi-Actor Regional Groups (MARGs) to lead co-creation activities in the establishment of eight bioeconomy-oriented regional HUBs in Campania (Italy), Peloponnese (Greece), Andalusia and Asturias (Spain), Centro (Portugal), Slovenia, Nouvelle-Aquitaine (France), and Cyprus. These HUBs will promote the exchange of best practices, and foster innovative methodologies for assessing environmental impacts and circularity in bio-based sectors. BioINSouth aims to enhance regional competitiveness, innovation capacity, and contribute to the EU's fair and green transition. A key component of BioINSouth is the analysis of regional stakeholders. Using the digital tool Wheesbee, developed by Innovation Engineering (PNO Group), the project determined bioeconomy technologies and top innovators based on their participation in funded projects over the last 10 years. This tool helped map stakeholders in BioINSouth HUBs regions, understanding the interests, objectives, and relationships of local actors, for developing effective bioeconomy strategies. By engaging diverse stakeholders, including policymakers, public authorities, market operators, civil society, and research organizations, the project ensures that methodologies and tools are tailored to regional needs and challenges. The stakeholder analysis identifies opportunities and barriers, enabling targeted interventions that drive sustainable bioeconomy practices. The overall methodology promotes collaboration and the exchange of best practices within a macro-regional BioINSouth Network and with international organizations. Demonstrated in eight regions, the concept will be transferable and replicable throughout other Southern European regions and beyond, ensuring sustainability and circularity in bio-based sectors. This project underscores the importance of regional policies in driving sustainable bioeconomy practices and fostering inclusive growth. Acknowledgment: Project BioINSouth (Grant Agreement No. 101156363) is co-funded by the European Union and supported by the Circular Bio-based Europe Joint Undertaking and its members. Views and opinions expressed are however those of the author(s) only and do not necessarily reflect those of the European Union or CBE JU. Neither the European Union nor the CBE JU can be held responsible for them.