Sustainable production of functional human serum albumin in Nicotiana benthamiana: a green biomanufacturing approach

Human serum albumin (HSA) is widely used in medicine and biotechnology, yet its demand exceeds plasma-derived supply. Recombinant production in microbial and mammalian systems presents cost and sustainability limitations. This study optimized transient expression of recombinant HSA (rHSA) in Nicotiana benthamiana and evaluated its ecological impact. The highest yield of 30 µg/g fresh weight was achieved at 4 days post-infiltration using Agrobacterium OD600 of 0.2 and 21-day-old seedlings. Purified rHSA exhibited 95% purity and significantly enhanced NIH3T3 fibroblast proliferation in a dose-dependent manner. Sustainability assessment using ComplexMoGAPI indicated moderate solvent efficiency (0.083 L/mg) and energy consumption (6.73 kWh/mg) but identified waste generation (9350 g/mg) and process mass intensity (10,166.67 g/mg) as major environmental constraints. While N. benthamiana offers a scalable rHSA production platform, improvements in waste valorization and solvent recovery are needed to enhance environmental performance. This study advances plant-based biomanufacturing as a sustainable strategy for biopharmaceutical production.