Lentinus subnudus was picked in the wild during the rainy season in the month of June and then transferred aseptically onto Potato Dextrose Agar (PDA) slant by tissue culture protocol. The optimal culture conditions for biomass and exopolysaccharide production was investigated. Growth requirements of the fungus were optimized for carbon, nitrogen, carbon-nitrogen ratio, seed culture volume and initial pH. Temperature condition was maintained at 25oC at 100 rpm for 5 days. Each of the culture__ampersandsignnbsp; arameters investigated gave significant increament on biomass and exopolysaccharide production. For Coptimization,the highest biomass and exopolysaccharide was produced by starch (6.56gl-1, 1.84gl-1) followed by glucose (2.01gl-1, 3.28gl-1) respectively. Tryptone and peptone N2-sources gave the highest biomass and exopolysaccharides as 2.44gl-1 and 3.06gl-1 respectively. The optimal C:N ratio for biomass and exopolysaccharides was found to be 1:? (2.75gl-1) and 1:1/5 (5.26gl-1) respectively. Highest biomass and exopolysaccharides were also produced at pH 8.5 (1.90 gl-1) and pH 5.5 (2.00gl-1) respectively. There was no significant difference in the biomass and exopolysaccharides produced when different volume of the substrate was used. Mutants and hybrid strains of L. subnudus were produced by exposure to ultraviolet radiation and by crossbreeding with L. edodes respectively. Three (3) of the mutants that gave higher yield and better performance were selected for further biochemical analyses and then compared with the wild and hybrid varieties. Biochemical analyses such as total protein, total sugar, total phenol, total ascorbic acid content, reducing and non-reducing sugar, starch contents, anti-oxidant capacities of the wild, mutants and hybrid were performed. From our result, the mutants gave higher production and performance as compared to the wild and hybrid strains.