Improving 10-deacetylbaccatin III-10-β-O-acetyltransferase catalytic fitness for Taxol production

Nat Commun. 2017 May 18:8:15544. doi: 10.1038/ncomms15544.

Abstract

The natural concentration of the anticancer drug Taxol is about 0.02% in yew trees, whereas that of its analogue 7-β-xylosyl-10-deacetyltaxol is up to 0.5%. While this compound is not an intermediate in Taxol biosynthetic route, it can be converted into Taxol by de-glycosylation and acetylation. Here, we improve the catalytic efficiency of 10-deacetylbaccatin III-10-O-acetyltransferase (DBAT) of Taxus towards 10-deacetyltaxol, a de-glycosylated derivative of 7-β-xylosyl-10-deacetyltaxol to generate Taxol using mutagenesis. We generate a three-dimensional structure of DBAT and identify its active site using alanine scanning and design a double DBAT mutant (DBATG38R/F301V) with a catalytic efficiency approximately six times higher than that of the wild-type. We combine this mutant with a β-xylosidase to obtain an in vitro one-pot conversion of 7-β-xylosyl-10-deacetyltaxol to Taxol yielding 0.64 mg ml-1 Taxol in 50 ml at 15 h. This approach represents a promising environmentally friendly alternative for Taxol production from an abundant analogue.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyltransferases / metabolism
  • Alanine / chemistry
  • Antineoplastic Agents / chemistry
  • Catalysis
  • Catalytic Domain
  • Glycosylation
  • Hydrogen-Ion Concentration
  • Magnetic Resonance Spectroscopy
  • Molecular Docking Simulation
  • Mutagenesis
  • Mutation
  • Paclitaxel / biosynthesis*
  • Paclitaxel / chemistry*
  • Plant Extracts
  • Recombinant Proteins / metabolism
  • Taxoids / chemistry*
  • Taxoids / metabolism*
  • Taxus / chemistry
  • Taxus / enzymology*
  • Temperature

Substances

  • 7-xylosyl-10-deacetyltaxol
  • Antineoplastic Agents
  • Plant Extracts
  • Recombinant Proteins
  • Taxoids
  • 10-deacetylbaccatine III
  • Acetyltransferases
  • Alanine
  • Paclitaxel