Difference between revisions of "Team:TU Darmstadt/Demonstrate"

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Revision as of 23:08, 29 October 2017

MainPage

Proof of Concept

Here we want to give you a short tour about what we achieved in the project. We show our results and give a view on the resulting product, a enzyme sensing hydrogel.

Chitin Synthase NodC

The functionality of the NodC enzyme was verified by performing the UDP-Glo™ Glycosyltransferase Assay. The evaluation of the assay shows that the NodC enzyme converts the UDP-GlcNAc to free UPD and a growing oligo-GlcNAc-chain. The free UDP is converted to ATP, which acts as a substrate for a luciferase reaction and creates luminescence. So the assay and the increasing luminescence depending on increasing enzyme concentrations shows that the NodC enzyme can create chitin oligomers.
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Figure 1. Activity assay of NodC. The NodC (40 ng) was titrated in 1X glycosyltransferase reaction buffer the presence of 100 μM of UDP-N-acetylglcosamine and 10 mM N-acetylglucosamine (GlcNAc) as an acceptor substrate. The reaction was performed as described before and the luminescence was measured after 1 hour of incubation with a Tecan200 Infinite Pro plate reader. Each point is an average of two experiments, and the error bars represent the standard deviations. RLU = relative light units.

Chitin Deacetylase NodB

Under Construction
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Figure 2.

Hydrogel

We manufactured several hydrogels containing non-toxic and cost-effectively gelling agent to form optimal wound dressings. Our hydrogels could be formed with basic laboratory equipment at any shape and could easily be adjust to the affected tissues.
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Figure 3. We manufactured a agar-chitosan hydrogel

Chemistry

We manufactured a protease-sensing chitosan derivative. For the verifaction of its functionality it has been measured via flourimeter. It showed a peak at 390 nm before the protease was added. After the cleavage with the protease a shift of this peak to the wavelength of 450 nm was observed.
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Figure 4. Fluormetric measurement of the peptide cleavage via proteases.