Expression of heparan sulfate sulfotransferases in Kluyveromyces lactis and preparation of 3'-phosphoadenosine-5'-phosphosulfate

Glycobiology. 2011 Jun;21(6):771-80. doi: 10.1093/glycob/cwr001. Epub 2011 Jan 11.


Heparan sulfate (HS) belongs to a major class of glycans that perform central physiological functions. Heparin is a specialized form of HS and is a clinically used anticoagulant drug. Heparin is a natural product isolated from pig intestine. There is a strong demand to replace natural heparin with a synthetic counterpart. Although a chemoenzymatic approach has been employed to prepare synthetic heparin, the scale of the synthesis is limited by the availability of sulfotransferases and the cofactor, 3'-phosphoadenosine-5'-phosphosulfate (PAPS). Here, we present a novel method to produce secreted forms of sulfotransferases in the yeast cells, Kluyveromyces lactis. Five sulfotransferases including N-sulfotransferase, 2-O-sulfotransferase, 3-O-sulfotransferase 1 and 6-O-sulfotransferases 1 and 3 were expressed using this method. Unlike bacterial-expressed sulfotransferases, the yeast proteins can be directly used to modify polysaccharides without laborious purification. The yeast-expressed sulfotransferases also tend to have higher specific activity and thermostability. Furthermore, we demonstrated the possibility for the gram-scale synthesis of PAPS from adenosine 5'-triphosphate at only 1/5000th of the price purchased from a commercial source. Our results pave the way to conduct the enzymatic synthesis of heparin in large quantities.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Carbohydrate Conformation
  • Gene Expression
  • Kluyveromyces / enzymology*
  • Phosphoadenosine Phosphosulfate / biosynthesis*
  • Phosphoadenosine Phosphosulfate / chemistry
  • Phosphoadenosine Phosphosulfate / isolation & purification
  • Polysaccharides / biosynthesis
  • Polysaccharides / chemistry
  • Sulfotransferases / biosynthesis*
  • Sulfotransferases / isolation & purification
  • Sulfotransferases / metabolism


  • Polysaccharides
  • Phosphoadenosine Phosphosulfate
  • Sulfotransferases
  • heparitin sulfotransferase