The invention provides an extraction method of soil protein. The extraction method comprises the following steps of adding soil protein extraction buffer liquid into a soil sample, oscillating, and standing; putting into boiling water for heating, cooling in 0 DEG C ice water, oscillating, and centrifuging; filtering supernatant, adding Tris saturated phenol into the filtered supernatant, oscillating, centrifuging, removing the supernatant, and collecting lower phenol solution; adding an ammonium acetate methanol solution into the lower phenol solution, and storing for 6 to 8h at the temperature of -20 DEG C; centrifuging, removing out the supernatant, collecting protein precipitate, cleaning the protein precipitate by acetone, and freeze-drying, so as to obtain protein dry powder. After detecting by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gelelectrophoresis) electrophoresis, compared with the protein band obtained by other two representative extraction methods of the soil protein, the band of the soil protein extracted by the method is clearer.