Abstract: The secondary metabolites of actinomycetes are rich and diverse, which have become important sources of antibiotics and their lead compounds. The study of microbial secondary metabolism is focused on metabolic regulation of antibiotic biosynthesis in actinomycetes by genetic manipulation. On the basis of the advances of recent years, this paper summarizes the application progress of metabolic regulation used for improving biosynthesis of antibiotics in actinomycetes, concluding of regulating the expression of regulatory genes, increasing the copy numbers of gene clusters and heterologous expression, over-expressing the resistance genes and transfer gene, improving precursor metabolic flux, and ribosome engineering.