Abstract: Mushroom has been valued throughout the world as a source of both food and medicine for thousands of years. The objective of this study has focused on the edible mushrooms (Pleurotus ostreatus and Agaricus bisporus) preparation of chitin nanowhiskers from mushrooms. Chitin nanowhiskers were prepared from both the mushrooms by acid hydrolysis method after the removal of contaminating proteins and other minerals. The concentration of acid for hydrolysis was standardized for the two mushrooms. Button mushroom has yielded maximum chitin (14.3%) when 9N HCl was used and the oyster mushroom showed maximum chitin yield (44.45%) when 12N HCl was used for hydrolysis. Thickness, length and surface characteristics of the chitin were assessed through SEM. Size of the nanowhiskers of the mushrooms ranged from 50-200nm in length and ~11nm of thickness.