AknK is an L-2-deoxyfucosyltransferase in the biosynthesis of the anthracycline aclacinomycin A

Biochemistry. 2004 Apr 20;43(15):4548-58. doi: 10.1021/bi035945i.


The antitumor drug aclacinomycin A is a representative member of the anthracycline subgroup that contains a C(7)-O-trisaccharide chain composed of L-2-deoxysugars. The sugar portion of the molecule, which greatly affects its biological activity, is assembled by dedicated glycosyltransferases; however, these enzymes have not been well-studied. Here we report the heterologous expression and purification of one of these enzymes, AknK, as well as the preparation of dTDP-L-2-deoxysugar donors, dTDP-L-2-deoxyfucose and dTDP-L-daunosamine, and the monoglycosyl aglycone, rhodosaminyl aklavinone. Our experiments show that AknK catalyzes the addition of the second sugar to the chain, using dTDP-L-2-deoxyfucose and rhodosaminyl aklavinone, to create the L-2-deoxyfucosyl-L-rhodosaminyl aklavinone. AknK also accepts an alternate dTDP-L-sugar, dTDP-L-daunosamine, and other monoglycosylated anthracyclines, including daunomycin, adriamycin, and idarubicin, to build alternate disaccharides on variant anthracycline backbones. Remarkably, AknK also catalyzes a tandem addition of a second L-2-deoxyfucosyl moiety, albeit with reduced activity, to the natural disaccharide chain to produce L-deoxyfucosyl-L-deoxyfucosyl-L-rhodosaminyl aklavinone, a variant of the natural aclacinomycin A. These results demonstrate that AknK may be a useful enzyme for the chemoenzymatic synthesis of anthracycline variants.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aclarubicin / analogs & derivatives
  • Aclarubicin / biosynthesis*
  • Catalysis
  • Cloning, Molecular
  • Fucose / analogs & derivatives
  • Fucose / biosynthesis
  • Fucose / chemistry
  • Fucosyltransferases / biosynthesis
  • Fucosyltransferases / genetics
  • Fucosyltransferases / isolation & purification*
  • Glycosylation
  • Hexosamines / biosynthesis
  • Hexosamines / chemistry
  • Naphthacenes / chemical synthesis
  • Streptomyces / enzymology*
  • Streptomyces / genetics
  • Substrate Specificity
  • Thymine Nucleotides / chemical synthesis
  • Trisaccharides / chemistry


  • Hexosamines
  • Naphthacenes
  • Thymine Nucleotides
  • Trisaccharides
  • aklavinone
  • daunosamine
  • Fucose
  • Aclarubicin
  • Fucosyltransferases