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

ABSTRACT: The natural concentration of the anticancer drug Taxol is about 0.02% in yew trees, whereas that of its analogue 7-?-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-?-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 ?-xylosidase to obtain an in vitro one-pot conversion of 7-?-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.

SUBMITTER: Li BJ 

PROVIDER: S-EPMC5454391 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature