The invention provides a high-efficiency soil protein extraction method, comprising the following steps: adding soil in a pre-cooling soil protein extraction buffer solution and blending and oscillating the mixture in a 4 DEG C shaking table over night; performing drawing and filtering of the shaken mixture in a buchner funnel using a crude filter paper; processing the drawing and filtering liquid by macroporous resin (D101), filtering the processed liquid through a 0.22 Mum microporous membrane, the filtered liquid being the extracellular protein mother liquid; processing the cells on the filter membrane by cleaning, crushing, centrifugalizing and vacuum drying to obtain the dried powder, namely intracellular soil protein sediment. The extracellular protein mother liquid is processed by centrifuging, separating and vacuum drying to obtain the intracellular soil protein dried powder; degrading the intracellular soil protein dried powder until the degraded intracellular soil protein dried powder is clearly separated on 10% polyacrylamide gel SDS-PAG electrophoresis, the method has high separation efficiency, microbial yield is 37.5%, which is higher than the yield of the traditional method 0.1-1%, simple operation flow, good SDS-PAGE resolution and good mass spectrum identification effect.