The production of bio-succinic acid (bio-SA) from biomass has the potential to partially replace some petrochemicals, reduce climate change by capturing carbon dioxide, and provide a cleaner environment by managing waste streams. This study evaluates the economics, environmental impact, risk assessment, and optimal processing route of bio-SA production from multiple feedstocks (first, second, and third-generation), including (1) glucose, (2) corn stover, (3) glycerol, and (4) seaweed. A superstructure-based optimization model consisting of 39 processing alternatives with a technology readiness level of 7–9 is developed, and the optimal topology for bio-SA production by maximization of the net present value under deterministic and stochastic conditions is identified. Once optimization is completed, the framework provides clear guidance for multi-criteria analysis, including the technical, economical, and environmental aspects of the biorefinery. The results indicate that glycerol is the best feedstock and corn stover is the second to best, producing bio-SA at selling prices of 1.6–1.9 USD per kg and 1.7–2.0 USD per kg, respectively, through their optimal processing pathways. Saccharina japonica (seaweed) is less suitable for large-scale bio-SA production due to the high cost of seaweed and the inability of enzymes to hydrolyze alginate, which is one of the major carbohydrate fractions (25–30 wt%) of this feedstock. The environmental results indicate that the optimal pathway from glycerol is the most environmentally friendly process, followed by optimal processing pathways from substrates such as corn stover, glucose, and S. japonica.