Extracting extracellular polymeric substances from fungi in contrasts: from quantity to quality

Many fungi are able to produce extracellular polymeric substances (EPS) for environmental, food, and industrial applications. This study evaluated the extraction (in vivo) of EPS from Rhodotorula mucilaginosa, a typical yeast with abundant EPS. Three extracting methods were set, i.e., heating, addition of NaCl during heating, and cation exchange resin (CER). The abundance of extracted proteins and polysaccharides showed evident contrasts (elevated to ~ 600 and 1700 mg/L, respectively) after heating at 70 °C in water. Although the higher temperature will increase the extracted abundance of EPS, the leakage of DNA would be enhanced due to cell rupture. The addition of NaCl further promoted the efficiency of extraction, either for proteins (from ~ 550 to ~ 650 mg/L) or polysaccharides (from ~ 1700 to ~ 2010 mg/L). Moreover, the biochemical results showed that the extracted abundance of EPS via heating was dramatically higher than that via CER. Additionally, DNA leakage in the CER treatment (2.0 g/g DW) was significantly higher (up to > 6 mg/L) than that under heating at 70 °C (< 2 mg/L). Furthermore, the three-dimensional excitation-emission matrix spectra showed two characteristic peaks of emission/excitation wavelength at 280/300 and 280/350, suggesting the relative high diversity of organic matters in EPS after heating treatments. Finally, a fluctuation of polysaccharide abundance in EPS at 500-1500 mg/L Pb²⁺ level was elucidated by the extraction based on heating treatment. This study hence confirmed that the heating method might be recommended for extraction of EPS from fungi in vivo KEY POINTS: • 3D-EEM results indicated that heating could extract more EPS compared with CER. • Heating treatments showed lower DNA leakage from fungi than CER treatments. • Addition of NaCl promoted the detachment of EPS from fungal cells in vivo.