Identification and characterization of so far undiscovered enzymes from secondary metabolite pathways

Project description

We are interested in exploiting the powerful reactivity and selectivity of enzymes from secondary metabolite pathways for the production of natural products and their analogs for pharmaceutical applications.

The hapalindole-type family of natural products is a fascinating group of compounds, produced solely by members of the Subsection V cyanobacterial strains. These hapalindole-type natural products exhibit several bioactivities of broad pharmaceutical interest, ranging from antituberculosis activity as well as dual-functional antitumor and antimitotic activity with an ability to reverse multidrug resistance. Hapalindoles are diversified into the welwitindolinones, fischerindoles and ambiguines through the employment of different enzymes such as oxygenases, methyltransferases and prenyltransferases. Despite the identification of the biosynthetic gene clusters for a number of cyanobacterial strains producing these compounds, not much is known about Rieske oxygenases involved in the biosynthesis of these compounds.

These enzymes belong to a fascinating class of biocatalysts with unprecedented catalytic capabilities that thus constitute an important tool in the synthesis and diversification of bioactive compounds.

The goal of the project is to shed light on the natural activity of these enzymes and to elucidate their potential as promising biocatalysts by investigating promiscuous activities of these enzymes.

The student will start with the expression of several Rieske oxygenases from the welwitindolinones gene cluster in E. coli. The expression levels will be investigated and, if necessary, optimized. In the second part of the project, organic extracts of cultured cyano- bacteria will be used to explore the natural activity of the enzymes by using HPLC-MS, NMR and GC. Finally, the substrate (and reaction) scope will be further elucidated by applying non-natural substrates.

Organization

The master research project will be conducted in the Department of Chemical and Pharmaceutical Biotechnology, in the research group of Dr. Sandy Schmidt. Our laboratory is well equipped for molecular biology, enzymology, analytics, and biocatalysis work. We offer a stimulating research environment for a student interested in these topics.

Contact

For more information, please contact Dr. Sandy Schmidt.
Chemical and Pharmaceutical Biology, GRIP
Building 3215, room 9.29
Antonius Deusinglaan 1
9713 AV Groningen