The present invention provides a method for producing and purifying β-carotene by Blakeslea trispora fermentation and use thereof. The method comprises the following steps: a) separately inoculating the Blakeslea trispora strains onto a PDA culture medium so as to obtain a spore suspension； b) propagating spores in a seeding tank so as to obtain seeds for fermentation； c) inoculating the seeds for fermentation onto a fermenter and fermenting said seeds； d) adjusting the fermentation liquid to be basic by using an organic or inorganic base, and filtering so as to obtain wet mycelia； e) treating the wet mycelia with a hydrophobic non-polar organic solvent； f) mixing the wet mycelia with an organic solvent of ester and obtaining a concentrated solution by extracting； g) adding a saturated monohydric alcohol into the concentrated solution, and filtering and crystallizing so as to obtain pure ss-carotene. The content of the ss-carotene in the present invention exceeds 96%, and the yield is above 85%.
