The invention provides a preparation method of microbial cellulose. The method comprises the following steps of: preparing a fermentation medium, namely sterilizing 5 percent of corn hydrolyzing sugar, 0.5 percent of ammonium sulfate, 0.5 percent of NaCl, 0.2 percent of magnesium sulfate, 0.1 percent of acetic acid and 0.15 percent of lactate for 10 to 30 minutes at the temperature of between 110 and 150 DEG C, cooling the mixture to 30 DEG C, and inoculating 4 percent of ultraviolet mutagenesis acetobacter aceti seed liquor onto the mixture, and culturing the mixture for 120 hours in a shake bed; after centrifuging the fermentation liquor for 20 minutes at the speed of 3,000r/min in a high-speed centrifugal machine, pouring the supernatant fluid; heating the centrifugal precipitate for 30 minutes in a boiling water bath of which the temperature is 100 DEG C by using 4 percent of Na OH solution, to remove mycoprotein; centrifuging the solution, adding a proper amount of neutral detergent solution into the precipitate, and heating the precipitate for 1 hour in the boiling water bath of which the temperature is 100 DEG C to remove impurities, such as impure sugars, lipoids and the like from the precipitate; repeatedly washing the precipitate with deionized water and 0.5 percent of acetic acid; and drying the precipitate for 24 hours at the temperature of 80 DEG C to obtain the microbial cellulose.