The invention discloses a culture medium added with a surfactant and application thereof. The culture medium added with the surfactant is composed of glycerinum or glucose which is taken as a major carbon source, a yeast extract and / or sodium glutamate which are / is taken as a major nitrogen source, inorganic salt components comprising Na2SO4, KH2PO4, K2HPO4, K2SO4 and MgSO4.7H2O, vitamins VB1, VB6 and VB12, and Tween 20, Tween 40, Tween 60 or Tween 60 added separately, wherein the concentration of the glycerinum or the glucose is 10-120g / L, and the concentration of the yeast extract and / or the sodium glutamate is 5-50g / L. The surfactant of an appropriate concentration is added to the culture medium so that the cell membrane permeability of microorganisms is changed, mass transfer of a substrate and oxygen is strengthened and growth is promoted; due to the adopted technical scheme, the conversion rate of the substrate can be increased without extra labor or equipment investment, and therefore, the fermentation of schizochytrium limacinum is effectively reduced.