The invention relates to a culture medium for clean production of riboflavin and a method. Molasses is replaced with glycine betaine, the matching proportion of glucose to molasses is regulated and controlled in basic materials and a fermentation process supplementary material culture medium, residues are reduced, and clean production is achieved. Meanwhile, traditional organic nitrogen source corn steep liquor and yeast powder are replaced with a novel organic nitrogen source baker's yeast extractum, metabolism regulation and bioengineering optimization are carried out, therefore, the biomassis increased, the fermentation period is shortened, and continuous beneficiating is achieved. By means of the method, fast growth of thalluses can be promoted, the specific growth rate can be increased, the thalluses are prompted to synthesize riboflavin efficiently, the fermentation culture time is shortened, and is shortened by 10 h to 20 h compared with a conventional process, the fermentationlevel is improved, in-tank residues are reduced (amino nitrogen is reduced by 52% to 68%), and the light-previous degree of fermented mash is improved by 17% to 20%; the aim of clean production is achieved, the production comprehensive cost is reduced, and the fine regulation and control are achieved.