Difference between revisions of "Part:BBa K729002"
Sednanalien (Talk | contribs) (→Results) |
(→Characterisation) |
||
| Line 22: | Line 22: | ||
===Characterisation=== | ===Characterisation=== | ||
| − | Plastic degradation is mediated via a laccase protein. As such, we will be using a laccase enzymatic activity assay to determine | + | Plastic degradation is mediated via a laccase protein. As such, we will be using a laccase enzymatic activity assay to determine it's production. Laccase catalyses the oxidation of <span class="footnote" title="Syringaldazine">syringaldazine</span>, a reaction that exhibits an observable OD change at 530nm. A sample of syringaldazine can be used as a blank in a spectrophotometer, against a sample containing syringaldazine and our laccase sample, allowing the rate of oxidation to be measured, and hence the enzymatic activity of laccase. |
In order to determine the effectiveness of laccase in degrading plastic, we will expose strips of various types of plastic to the laccase expressing bacteria, before viewing the strips under a scanning electron microscope. This will allow us to compare the pitting in plastic samples treated with laccase, to the untreated samples, allowing us to determine the extent of plastic degradation. | In order to determine the effectiveness of laccase in degrading plastic, we will expose strips of various types of plastic to the laccase expressing bacteria, before viewing the strips under a scanning electron microscope. This will allow us to compare the pitting in plastic samples treated with laccase, to the untreated samples, allowing us to determine the extent of plastic degradation. | ||
Revision as of 11:21, 21 September 2012
Laccase for Polyethylene Degradation
Laccase for the degradation of polyethylene and organic pollutants. Sequence pending.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 225
- 1000COMPATIBLE WITH RFC[1000]
Description
Characterisation
Plastic degradation is mediated via a laccase protein. As such, we will be using a laccase enzymatic activity assay to determine it's production. Laccase catalyses the oxidation of syringaldazine, a reaction that exhibits an observable OD change at 530nm. A sample of syringaldazine can be used as a blank in a spectrophotometer, against a sample containing syringaldazine and our laccase sample, allowing the rate of oxidation to be measured, and hence the enzymatic activity of laccase.
In order to determine the effectiveness of laccase in degrading plastic, we will expose strips of various types of plastic to the laccase expressing bacteria, before viewing the strips under a scanning electron microscope. This will allow us to compare the pitting in plastic samples treated with laccase, to the untreated samples, allowing us to determine the extent of plastic degradation.
Results
Our results indicate a significantly higher rate of oxidation for our BioBrick than the control. This indicates that our transformed E. Coli have successfully produced laccase, allowing it to oxidise the syringaldazine utilised in the laccase assay.