This study was performed to investigate the usability of chicken feather hydrolysate (Chicken feather peptone (CFP)) as substrate for mycelial biomass and extracellular polysaccharides (EPS) production from edible mushroom Morchella esculenta. The ability of CFP to support biomass and EPS production in edible mushroom M. esculenta was compared to those of two commercial peptones (Tryptone peptone (TP) and Fish peptone (FP)). The maximum biomass (16.3 g/l) and EPS (4.8 g/l) concentrations were achieved with TP. Second, high biomass (15.9 g/l) and EPS (4.6 g/l) concentrations were obtained with CFP. Also, biomass and EPS concentrations in CFP medium were statistically near to those in the TP medium. CFP and TP resulted in not only uniform pellets with smaller size (5 mm) but also faster mycelial growth compared to FP. This study showed for the first time that CFP could be effectively used as a novel EPS production substrate.