Difference between revisions of "Part:BBa K620001"
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[[Image:Blank BPA-2.jpg|frame|center|HPLC of BPA, without enzyme added]] | [[Image:Blank BPA-2.jpg|frame|center|HPLC of BPA, without enzyme added]] | ||
[[Image:WT-F87A-2.jpg|frame|center|HPLC of BPA degraded, with p450 Wt-F87A]] | [[Image:WT-F87A-2.jpg|frame|center|HPLC of BPA degraded, with p450 Wt-F87A]] | ||
− | As shown in the HPLC results, the[[:Image: | + | As shown in the HPLC results, the[[:Image:Blank BPA-2.jpg| plain BPA HPLC]] has only one peak, while the [[:Image:WT-F87A-2.jpg|HPLC analysis of BPA degraded with the WT-F87A cytochrome p450]] has two. |
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> |
Latest revision as of 03:55, 29 September 2011
WT-F87A (p450)
A wild-type p450 with a mutation at the 87th amino acid. It is confirmed to degrade bisphenol A (BPA), but as it is a promiscuous protein, it can likely bind and degrade other organic molecules.
Usage and Biology
This image shows the GCMS of BPA before degradation, in which the mass of BPA (227g/mole) is clearly visible.
This image shows BPA after a reaction was conducted with this p450. The p450 was expressed in cells and extracted. Then a reaction was prepared with 5 microliters of 20mM BPA in DMS0, 12 microliters of a 5 micromolar p450 solution, 20 microliters glucose, and 2 microliters GDH, in 161 microliters 1M KPI buffer. This reaction was performed for 4 hours and analyzed using GCMS. Here, there is an apparent large 205g/mole peak, and the 227 g/mole peak is greatly reduced. This indicates that BPA is being reduced to several structures weighing 205 g/mole.
As shown in the HPLC results, the plain BPA HPLC has only one peak, while the HPLC analysis of BPA degraded with the WT-F87A cytochrome p450 has two. Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 75
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 1219
Illegal SapI.rc site found at 2242
Illegal SapI.rc site found at 2842