The effect of medium components (carbon, nitrogen, and mineral sources) and environmental factors (initial pH and temperature) for mycelial growth and exopolysaccharide (EPS) production in Sarcodon aspratus(Berk) S.lto TG-3 was investigated. The optimal temperature (25°C) and initial pH (5.0) for the EPS production in shake flask cultures of S. aspratus were determined using the two-dimensional contour plot. The most suitable carbon, nitrogen, and mineral sources for EPS production were glucose, yeast extract, CaCl2 and KH2PO4, respectively. Notably, the EPS production was significantly enhanced by supplementation of calcium ion. Subsequently, the optimum concentration of glucose (30gl−1), yeast extract (15gl−1), CaCl2 (1.1gl−1), and KH2PO4 (1.2gl−1) were determined using the orthogonal matrix method. The effects of nutritional requirement on the mycelial growth of S.aspratuswere in regular sequence of glucose>KH2PO4>yeast extract>CaCl2, and those on EPS production were in the order of glucose>yeast extract>CaCl2>KH2PO4. Under the optimal culture conditions, the maximum EPS concentration in a 5-l stirred-tank reactor was 2.68gl−1 after 4days of fermentation, which was 6-fold higher than that at a basal medium. The two-dimensional contour plot and orthogonal matrix method allowed us to find the relationship between environmental factors and nutritional requirement by determining optimal operating conditions for maximum EPS production in S.asparatus. The statistical experiments used in this work can be useful strategies for optimization of submerged culture processes for other mushrooms.
