Biochemical changes of polysaccharides and proteins within EPS under Pb(II) stress in Rhodotorula mucilaginosa

Microorganisms have been widely applied to heavy metal adsorption due to their strong secretion of extracellular polymeric substances (EPS). This study explored the responses of Rhodotorula mucilaginosa (R1, a red yeast with substantial EPS supply) under Pb stress. The maximum sorption of Pb cations by R1 was ~650 mg/L. In particular, despite the declined microbial biomass, the total Pb sorption after incubation was actually elevated in the solution with high Pb concentration. At 0-1000 mg/L Pb(NO₃)₂ level, the longitudinal sizes of the yeast capsules increased from 2.04 to 2.90 µm. At 1500 mg/L, however, the survived yeast started to lose the membrane integrity of the cells. Meanwhile, the percentages of organic carbon contents of EPS decreased from 40% to 33% when the Pb(NO₃)₂ concentration raised to 2500 mg/L, confirming the incorporation of Pb²⁺ cations into the fungal EPS during the sorption. For the survived R1 cells, function of polysaccharides to resist Pb toxicity only worked at extremely high Pb(NO₃)₂ levels (>= 1500 mg/L). In contrast, proteins showed continuously enhanced ability to resist Pb toxicity, consistent with their increasing content (per cell) in the EPS. Moreover, ATR-IR spectra showed that the intensity of amide II peak at 1540 cm^-1 was significantly increased, indicating elevated glutathione (GSH) in EPS. This suggested that GSH could be the critical Pb-binding component in EPS proteins. This study hence elucidated roles of polysaccharides and proteins in EPS under the toxicity caused by heavy metals.