Glycopeptidolipids (GPLs) attached to the outer surface of the greasy cell envelope, are a class of important glycolipids synthesized by several non-tuberculosis mycobacteria. The deletion or structure change of GPLs confers several phenotypical changes including colony morphology, hydrophobicity, aggregation, sliding motility, and biofilm formation. In addition, GPLs, particular serovar specific GPLs, are important immunomodulators. This review aims to summarize the advance on the structure, function and biosynthesis of mycobacterium GPLs. J. Cell. Biochem. 114: 1705–1713, 2013. © 2013 Wiley Periodicals, Inc.