The invention relates to a PU adhesive for single-component microfiber leather and a preparation method of the PU adhesive. The preparation method comprises the following steps: adding polyester polyol into a reaction bottle, heating to melt the polyester polyol, adding an antioxidant BHT and phosphoric acid, uniformly stirring, adding molten isocyanate, reacting at 110 DEG C, adding an organic solvent in the reaction process for dilution, and reacting until the viscosity of the system reaches 80,000-120,000 cps/DEG C; and adding another organic solvent into the reaction bottle so as to adjustthe solid content of the system to 40%, continuously stirring for 1 hour, and packaging. The PU adhesive for the single-component microfiber leather prepared by the scheme has the advantages of simple process, convenience in operation and the like; the prepared PU adhesive is low in softening point, has good hydrolysis resistance, heat resistance and mildew resistance and is suitable for dry sticking, and when the PU adhesive is used for microfiber veneering, the matte phenomenon is avoided.