<p>Lincomycin A and celesticetin are representative members of the lincosamide class of clinically used antibiotics produced by <i>Streptomyces</i> species. Their distinctive chemical architectures arise from atypical biosynthetic gene clusters that lack well observed signature genes, and since the complete determination of the lincomycin A biosynthetic pathway, current research has focused on the genetic manipulation of regulatory elements and the protein engineering of biosynthetic enzymes. This review summarizes recent advances in elucidating the transcriptional regulation of lincosamide biosynthetic gene clusters and the structure–function relationships and engineering of their biosynthetic enzymes.</p>