Soluble extracellular polymeric substances and microplastics: Exposure-response and circular reuse for removal

Microplastics (MPs) are pervasive in aquatic systems, threatening ecosystems, human health, and microalgal production. Soluble extracellular polymeric substances (S-EPS) can agglomerate particles and aid removal. This study examines S-EPS from the cyanobacterium Cyanocohniella rudolphia (BEA 0786B) to (i) model and optimise S-EPS production, (ii) assess production in water contaminated with polystyrene MPs (PS-MPs), and (iii) test S-EPS as a bioflocculant for PS-MPs removal. Response surface methodology (RSM) defined a cost-lean operating window and predicted an optimum S-EPS titre of 113 mg/L at 7 days using 10 g/L nitrogen, 0.98 g/L phosphorus, and a biomass-to-medium ratio of 1:6.87 (w/v). Cultures were challenged with PS-MPs (50 μg/L and 5 mg/L) under static or aerated conditions, and at both exponential and stationary phases, and showed stimulated S-EPS synthesis with increases of up to 34%, depending on hydrodynamics and growth stage. Purified S-EPS were evaluated as a bioflocculant at 2 g/L PS-MPs to probe robustness and rate-limiting mechanisms and to delineate a conservative operating window. Maximum removal of 82% was achieved in freshwater at pH 3.5 with Fe3+ 0.05% (w/w), 25 °C, S-EPS dose 400 mg/L (S-EPS:PS-MPs 1:5, w/w), and 60 min flocculation. Zeta potential trends and microscopy support charge neutralisation/bridging as the dominant mechanism. Compatible with standard coagulation/flocculation units, the approach links cost-lean, cultivation-derived S-EPS (typically discarded) to their reuse as a low-additive pretreatment for algal-cultivation intake waters (freshwater/low-salinity), reducing reliance on synthetic coagulants and added salinity/metal-sludge burdens. Overall, C. rudolphia is a promising S-EPS producer, whose production is enhanced by exposure to PS-MPs, and its S-EPS acts as an efficient, bio-based flocculant for PS-MPs. The results support process designs to safeguard microalgal operations and to mitigate microplastic pollution in water. This work integrates RSM-optimised S-EPS production, environmental-level exposure-response, and a high-load removal benchmark, enabling circular, low-additive, drop-in pretreatment compatible with standard coagulation/flocculation units.