Promoters from the itaconate cluster of Ustilago maydis are induced by nitrogen depletion

Thiemo Zambanini, Sandra K. Hartmann, Lisa M. Schmitz, Linda Büttner, Hamed Hosseinpour Tehrani, Elena Geiser, Melanie Beudels, Dominik Venc, Georg Wandrey, Jochen Büchs, Markus Schwarzländer, Lars M. Blank, Nick Wierckx

Ustilago maydis is known for its natural potential to produce a broad range of valuable chemicals, such as itaconate, from both industrial carbon waste streams and renewable biomass. Production of itaconate, and many other secondary metabolites, is induced by nitrogen limitation in U. maydis. The clustered genes responsible for itaconate production have recently been identified, enabling the development of new expression tools that are compatible with biotechnological processes. Here we report on the investigation of two of the native promoters, P tad1 and P mtt1 , from the itaconate cluster of U. maydis MB215. For both promoters the specific activation upon nitrogen limitation, which is known to be the trigger for itaconate production in Ustilago, could be demonstrated by gfp expression. The promoters cover a broad range of expression levels, especially when combined with the possibility to create single- and multicopy construct integration events. In addition, these reporter constructs enable a functional characterization of gene induction patterns associated with itaconate production. The promoters are well suited to induce gene expression in response to nitrogen limitation, coupled to the itaconate production phase, which contributes towards the further improvement of organic acid production with Ustilago.