Overexpression of Trp-related genes in Claviceps purpurea leading to increased ergot alkaloid production

Abstract

The parasitic fungus Claviceps purpurea has been used for decades by the pharmaceutical industry as a valuable producer of ergot alkaloids. As the biosynthetic pathway of ergot alkaloids involves a common precursor L-tryptophan, targeted genetic modification of the related genes may improve production yield. In this work, the S76L mutated version of the trpE gene encoding anthranilate synthase was constitutively overexpressed in the fungus with the aim of overcoming feedback inhibition of the native enzyme by an excess of tryptophan. In another approach, the dmaW gene encoding dimethylallyltryptophan synthase, which produces a key intermediate for the biosynthesis of ergot alkaloids, was also constitutively overexpressed. Each of the above manipulations led to a significant increase (up to 7-fold) in the production of ergot alkaloids in submerged cultures.