The invention provides a method for desulfurization regeneration of waste rubber by mixing two types of microorganism bacteria and relates to a desulfurization regeneration technology of the waste rubber. According to the method, an enzyme generated in the mixed growth process of sphingomonas sp. and gordonia sp. is used for performing desulfurization regeneration on the waste rubber. An early study shows that the sphingomonas sp. or the gordonia sp. has a good desulfurization effect on waste tread rubber, styrene-butadiene rubber and natural rubber, and in order to study mixed bacterial desulfurization, the system provided by the invention adjusts a culture medium for symbiosis of the two bacteria, and the two bacteria are mixed together to perform desulfurization regeneration on the waste rubber. A study finds that during co-culture desulfurization, the biomass of the mixture of the two types of bacteria is higher than that of a single strain, which shows that the two bacteria have a synergistic effect. According to the method provided by the invention, the inoculation means of the two types of bacteria and the influence of process conditions for growth and metabolism of the two types of bacterial on the desulfurization of the waste rubber are studied, so that a best process for co-culture desulfurization is determined. Compared with undesulfured waste rubber, the desulfurized rubber is increased in selling value, lowered in surface sulfur element content, and increased in oxygen content. Therefore, the mixing of the two types of the microorganism bacteria has a relatively obvious desulfurization effect on the rubber surface.