Difference between revisions of "Part:BBa K391006"

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==DspB: carbohydrate digesting enzyme==
 
==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 <i>Escherichia coli</i> and <i>Staphylococcus epidermidis</i>. According to literature, DspB effectively cleaves these bonds and impedes biofilm formation.
 
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 <i>Escherichia coli</i> and <i>Staphylococcus epidermidis</i>. According to literature, DspB effectively cleaves these bonds and impedes biofilm formation.
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== Characterization Data ==
 
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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 and the result is shown below.
 
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<div style="text-align:center;">[[Image:Igem2010_assay2_linegraph.jpg‎|500px]]</div>
 
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The graph above plots the absorbance at wavelength 405nm (y-axis) over time in days (x-axis). It can be observed from the graph that over time, the absorbance at 405nm increases with the treatment with the lysate containing DspB by a significant amount compared to the controls. This assay has been replicated and the results have shown to be reproducible.
 

Latest revision as of 07:14, 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.