Aims: The objective of the present study was to determine the optimal culture conditions for the production of four groups of exopolysaccharides (EPSs) in Phellinus gilvus by submerged culture and to investigate their molecular properties by multi-angle laser-light scattering (MALLS) analysis. Methods and Results: The optimal temperature and initial pH for the production of both mycelial biomass and EPSs in P. gilvus by submerged flask cultures were found to be 30degreesC and pH 9.0, respectively. Glucose and corn steep powder were the most suitable carbon and nitrogen source for both mycelial biomass and EPS production. Optimal medium composition was determined to be glucose 30 g l(-1) , corn steep powder 5 g l(-1) , MgSO4 1.23 g l(-1) , KH2 PO4 0.68 g l(-1) , and K-2 HPO4 0.87 g l(-1) . Four groups of EPSs (Fr-I, II, III, and IV) were obtained from the culture filtrates by gel filtration chromatography on Sepharose CL-4B and characterized by size exclusion chromatography (SEC) coupled with MALLS. The weight average molar mass (M (w) ) of Fr-I, Fr-II, Fr-III and Fr-IV were determined to be 8.628 x 10(6) (+/-129 420), 1.045 x 10(6) (+/-19 855), 61.09 x 10(4) (+/-1244), and 33.55 x 10(4) (+/-134) g mol(-1) , respectively. Conclusions: Under optimal culture conditions, the maximum EPS production in a 5-l stirred fermenter indicated 5.3 g l(-1) after 11 days of fermentation. The SEC/MALLS analysis revealed that Fr-I, which has extremely high molecular weight, was presumably an aggregate of complex polysaccharides forming a compact globular shape; whereas Fr-II was nearly spherical, Fr-III and Fr-IV were rod-like chains in an aqueous solution. Significance and Impact of the Study: This is the first report on the production of high amounts of EPSs from liquid-culture of the basidiomycete, P. gilvus . The SEC/MALLS approach used in this study could be useful in providing greater insight into the characterization of the mushroom polysaccharides without carrying out elaborate fractionation procedures prior to analysis.