The invention provides a preparation method of sargassum fusiforme dietary fiber. The method includes the steps of: drying, enzymolysis, acidolysis, alkaline hydrolysis, centrifugation, gelation, bleaching, drying, activation, alcohol dehydration and drying. The method provided by the invention employs a composite plant hydrolase to assist papain to extract sargassum fusiforme dietary fiber, greatly improves the protein hydrolysis rate of sargassum fusiforme, overcomes the disadvantage of poor one-time protein removal effect in common enzymatic method, effectively destroys cell walls, enhancesthe full permeation of a fiber crystalline area, completely releases part of the protein closely combined to residue, and is conducive to degradation of papain to protein so as to improve the productpurity.