Difference between revisions of "Part:BBa K391006"
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== Characterization Data == | == Characterization Data == | ||
| − | The activity of DspB was tested by taking a crude cell lysate from | + | The activity of DspB was tested by taking a crude cell lysate from </i>E.coli</i> 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. |
<div style="text-align:center;">[[Image:Igem2010_assay2_linegraph.jpg|500px]]</div> | <div style="text-align:center;">[[Image:Igem2010_assay2_linegraph.jpg|500px]]</div> | ||
Revision as of 05:35, 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 </i>E.coli</i> 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.
