A shoe sanitizing assembly includes a pan that is positionable adjacent to an entry of a building such that the pan can to be stepped in by patrons of the building. A plurality of biasing members is each coupled to and extends upwardly from the pan. A mat is positionable on top of the plurality of biasing members such that the patrons of the building step on the mat when the patrons pass through the entry. A pair of fluid reservoirs is provided that each contains a fluid and each of the fluid reservoirs is positioned in the pan. A plurality of pumps is each positioned in the pan. Each of the pumps is actuated when the mat is stepped upon to spray a measured amount of the fluid outwardly through the mat onto soles of shoes worn by the user.