The invention relates to soybean protein isolate powder as well as a preparation method and use thereof. The preparation method comprises the following steps: mixing low-temperature degreased bean pulp with water, carrying out shearing, adding phytase and polyphenol oxidase for reaction, carrying out solid-liquid separation, shearing obtained bean dregs, adding water for extraction, carrying out solid-liquid separation, and combining an obtained water extracted bean emulsion with soybean milk extracting liquid, so as to obtain mixed soybean milk; regulating the pH value of the mixed soybean milk to settle soybean protein, and carrying out solid-liquid separation, so as to obtain a fixed phase, namely the soybean acid settled milk protein; preparing protein liquid from the soybean acid settled milk protein, regulating the pH value to 2.5-3.5, adding acid proteinase for enzymolysis, and carrying out solid-liquid separation, so as to obtain protein liquid; and concentrating the protein liquid, and carrying out spray drying on the obtained soybean acid protein fluid, so as to obtain the soybean protein isolate powder. A solution obtained by adding the soybean protein isolate powder into an acidic beverage has relatively good appearance transparency and extremely good storage stability.