The invention relates to a preparation method of macroporous polyacryamide resin. The method comprises the following steps of: preparing an aqueous phase, preparing an oil phase, preparing the macroporous polyacryamide resin and carrying out after-treatment on products. The preparation method of the macroporous polyacryamide resin has simplicity and low reaction temperature, and the prepared macroporous polyacryamide resin has macroporous structure from the surface to the interior, uniform and dense aperture, relatively larger specific surface area and strong water absorbability and adsorbability, can be used as a carrier of an immobilized enzyme and can greatly improve the capacity of the enzyme.