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

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<figcaption style="width:80%;"> <b>Figure 1. Activity assay of NodC.</b> The NodC (40 ng) was titrated in 1X glycosyltransferase reaction buffer the presence of 100&nbsp;μM of UDP-<i>N</i>-acetylglcosamine and 10&nbsp;mM <i>N</i>-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 <i>Tecan200 Infinite Pro</i> plate reader. Each point is an average of two experiments, and the error bars represent the standard deviations. RLU = relative light units.
 
<figcaption style="width:80%;"> <b>Figure 1. Activity assay of NodC.</b> The NodC (40 ng) was titrated in 1X glycosyltransferase reaction buffer the presence of 100&nbsp;μM of UDP-<i>N</i>-acetylglcosamine and 10&nbsp;mM <i>N</i>-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 <i>Tecan200 Infinite Pro</i> plate reader. Each point is an average of two experiments, and the error bars represent the standard deviations. RLU = relative light units.
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Revision as of 21:19, 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.

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.