Difference between revisions of "Part:BBa K391006"

(Characterization Data)
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== Characterization Data ==
 
== Characterization Data ==
 
The activity of DspB was tested by taking a crude cell lysate from e.coli cells expressing the protein. The protein in the lysate was then tested against a chromogenic substrate: 4-Nitrophenyl N-acetyl-β-D-glucosaminide. As DspB cleaves this substrate, the absorbance shifts from 300nm (substrate: 4-Nitrophenyl N-acetyl-β-D-glucosaminide) to 405nm (product: 4-nitrophenol. We measured the change in the level of absorbance at 405nm over time.
 
The activity of DspB was tested by taking a crude cell lysate from e.coli cells expressing the protein. The protein in the lysate was then tested against a chromogenic substrate: 4-Nitrophenyl N-acetyl-β-D-glucosaminide. As DspB cleaves this substrate, the absorbance shifts from 300nm (substrate: 4-Nitrophenyl N-acetyl-β-D-glucosaminide) to 405nm (product: 4-nitrophenol. We measured the change in the level of absorbance at 405nm over time.
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<div style="text-align:center;">[[Image:Igem2010_assay2_linegraph.jpg|500px]]</div>

Revision as of 05:34, 27 October 2010

DspB: carbohydrate digesting enzyme

Dispersin B (DspB) is an enzyme that degrades biofilms by catalyzing the hydrolysis of poly-ß-(1,6)-linked N-acetylglucosamine bonds. These bonds exist in the extracellular polymeric substance (EPS), which acts as a polysaccharide adhesin relevant to biofilm formation and integrity in Escherichia coli and Staphylococcus epidermidis. According to literature, DspB effectively cleaves these bonds and impedes biofilm formation.


Characterization Data

The activity of DspB was tested by taking a crude cell lysate from e.coli cells expressing the protein. The protein in the lysate was then tested against a chromogenic substrate: 4-Nitrophenyl N-acetyl-β-D-glucosaminide. As DspB cleaves this substrate, the absorbance shifts from 300nm (substrate: 4-Nitrophenyl N-acetyl-β-D-glucosaminide) to 405nm (product: 4-nitrophenol. We measured the change in the level of absorbance at 405nm over time.

Igem2010 assay2 linegraph.jpg