Torben Nielsen

Elucidation of carbohydrate converting enzyme mechanisms using DNP-NMR

Christian Kjeldsen

Research focus
Having a better understanding of the mechanisms of carbohydrate converting enzymes will be imperative to the development of methods for transglycosylation using cheap sugars, e.g. lactose, a byproduct of the dairy industry, to enzymatically synthesize important useful and valuable carbohydrates such as probiotics.

DNP-NMR can give information about lowly populated intermediates and we plan to use this to try to observe real intermediates in the conversion of carbohydrates. The strength of the method is the possibility to observe experimentally lowly populated intermediates and maybe even transition states. In contrast to the investigations using inactive mutants or modified substrates, this will use active enzymes and natural substrates. The only requirement is the need for incorporation of 2H and possibly 13C in the carbohydrates. Some simple isotope labeled carbohydrates are commercially available, but also special structures needs to be synthesized as part of the project.

The project contributes to the HYPERMAG Center by gaining insight into the inner workings of enzymes in real time using hyperpolarized nuclear magnetic resonance and thus reaching a better understanding of the chemistry and biochemistry of enzymatic carbohydrate conversion.

Scientific output
Find Christian's publications at DTU's online research database ORBIT.

Funding
The project is funded by Novo Nordisk Foundation and DTU Chemistry. The project is part of the HYPERMAG Center of Excellence funded by Danish National Research Foundation (DNRF124).

Supervisor
Professor Jens Ø. Duus.

Project Period
October 2014 - October 2017