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Transitioning from a fossil economy to a bio-economy will inevitably increase the demand for biomass production. One strategy to meet the demand is to re-cultivate set-aside arable land. This study investigated the climate impact and energy potential of grass-based biogas produced using fallow land in Uppsala municipality, Sweden. The assessment was performed on regional level for more than 1000 individual sites, using the agro-ecosystem model DeNitrification DeComposition (DNDC) in combination with time-dynamic life cycle assessment methodology. The results showed that the system could significantly increase biogas production within the region, which would reduce the climate impact by 9950 Mg CO 2 -eq per year. Compared with diesel fuel, the grass-based biogas gave a GWP reduction of 85%. However, the site-specific GWP reduction showed large spatial variability, ranging between 102 and 79% compared with diesel fuel, depending on where in the region the grass was cultivated. Two alternative scenarios were investigated, increased mineral N fertilisation and inclusion of N-fixing crops in the feedstock mixture. The highest mitigation per biogas energy produced was found for the N-fixing scenario but, because of lower yields, this scenario had lower total mitigation potential for the region than the increased fertilisation scenario. The increased fertilisation scenario had a lower climate mitigation effect per biogas energy produced, but the highest mitigation potential when the whole region was considered, because of the increased biogas production. The method applied in this study can guide land-use planning of local energy production from arable land, also for other regions. • The grass-based biogas production significantly reduced the regional climate impact. • The climate impact showed large spatial variation within the region. • The most suitable locations, from a mitigation perspective, were identified. • Increasing fertilisation intensity increased total mitigation for the region. • N fixing feedstock resulted in the highest mitigation per MJ. [ABSTRACT FROM AUTHOR]