A method for effectively preparing a cornea acellular matrix tissue engineering scaffold by enzymatic method is as follows: (1) a porcine corneal matrix plate layer is put into diluted Myroilysin for decellularization, and flushed respectively with normal saline and distilled water to obtain a porcine corneal acellular scaffold board layer; (2) the porcine corneal acellular scaffold is put into a drill floor, a riboflavin solution is added dropwise, and the porcine corneal acellular scaffold is irradiated by an UV lamp, and then flushed respectively with the normal saline and the distilled water to obtain a riboflavin-crosslinked porcine corneal acellular scaffold; and (3) the riboflavin-crosslinked porcine corneal acellular scaffold obtained by the step (2) is dehydrated, sterilized and stored at 4 DEG C for standby use. The prepared acellular scaffold still retains enough mechanical strength, certain pores are formed, and after dehydration, the acellular scaffold has high transparency and is favorable for cell growth and migration.