The invention belongs to the technical field of industrial microorganisms, and relates to a method for producing biological de-emulsifying bacteria by using a surplus sludge ultrasonic extracting solution from a municipal domestic sewage plant, which comprises the following steps: taking surplus sludge from a sewage treatment plant, carrying out ultrasonic pretreatment, and carrying out solid-liquid separation to obtain a clear sludge extracting solution; and (2) after regulating the pH value of the sludge extracting solution and adding an additional carbon source, forming a complete culture medium, sterilizing, inoculating a biological de-emulsifying bacteria Alcaligenes sp. TR-1 complete culture solution, and fermenting and culturing to obtain the biological de-emulsifying bacteria Alcaligenes sp. TR-1.Compared with the prior art, the invention uses the sludge extracting solution to ferment and produce the biological de-emulsifying bacteria Alcaligenes sp. TR-1, thereby lowering the production cost; and compared with the inorganic salt culture medium culture process, the invention can effectively enhance the thallus yield and shorten the fermentation time, keeps the property de-emulsifying property of the obtained thallus at a high level, is beneficial to popularization of the biological de-emulsifier, and can provide a new way for decrement and recycling of surplus sludge.