Abstract: Lentinus edodes with rich nutrients and health benefits can be added into food as flavor substance and nutrition base, endowing food with nutritional and functional properties. Using plain flour and Lentinus edodes powder as main materials, we conducted four-factor five-level response surface experiment taking Lentinus edodes powder addition amount, blend oil addition amount, soft sugar addition amount and egg addition amount as factors on the basis of single factor experiments, and fuzzy evaluation to optimize the formula of Lentinus edodes biscuits. Results showed that the optimum formula was determined as follows: self-raising plain flour 75 g, Lentinus edodes powder 25 g, blend oil 51 g, soft sugar 43 g, egg 32 g, baking soda 1 g, and ammonium bicarbonate for food 1 g. The Lentinus edodes biscuits prepared according to the optimum formula had advantages of uniform color, aromatic flavor and crisp taste, and its sensory score was 76.75.