Riboflavin is an important nutrient for humans and animals. Industrial production has shifted completely from chemical synthesis to microbial fermentation. First generation riboflavin production was improved by a combination of traditional mutagenesis and genetic engineering, and isolated strains have been used in industry. As the DNA genome of riboflavin producers has the potential to reveal new technologies, DNA microarray, proteomic and metabolic analyses have been applied to the analysis of hyper-riboflavin producers. In this review, disparity mutagenesis technology is introduced as a means of improving riboflavin production by Ashbya gossypii. DNA microarray, proteomic and metabolic analyses of this high riboflavin producer are discussed, as well as recent riboflavin production trends, costs and future improvements.