Team:Copenhagen/Project

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<strong>Cytochrome p450</strong>

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Cytochromes p450 is one of the largest gene superfamilies coding for enzymes present in the genomes in all biological kingdoms [6]. The enzymatic activities of these proteins are extremely diverse with activity in biotransformation of drugs, bioconversion of xenobiotics, biosynthesis of compounds as steroids, fatty acids, eicosanoids, fat soluble vitamins and bile acids. Furthermore, cytochrome p450’s are involved in the conversion of alkanes, terpenes and aromatic compounds as well as degradation of herbecides and insecticides [6]. </p>

<p>We will exploit this activity of cytochromes p450 in the bioconversion of xenobiotics to neutralize damaging medical residues in waste water.</p>

<p>Cytochromes p450 are defined as heme-thiolate proteins featuring a particular spectral signature at 450 nm, thus the name [7]. Despite the label as cytochromes, these proteins are not involved in electron transfers, but act as monooxygenases in a wide range of reactions such as epoxidation, N-dealkylation, O-dealkylation, S-oxidation and hydroxylation [8]. </p>

<p>We will focus on the hydroxylating property of the cytochromes, which is also the defining reaction for these enzymes.  The reductive activation of molecular oxygen reduces one of the oxygen atoms to a molecule of water, as the other is inserted into the substrate [7].</p>

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<table class="https://static.igem.org/mediawiki/2011/c/cc/CYP_catalystisk_circle.PNG" align="center">

<caption align="bottom"><p align="justify"><b>Figure 1</b>The P450 catalytic cycle in hydroxylation</p></caption>

<tr><td><img src="https://static.igem.org/mediawiki/2011/c/cc/CYP_catalystisk_circle.PNG" width="500px"></td></tr>

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<strong>CyperMan - Saves the Fish</strong>

 

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We intend to transform E.coli bacteria with different combinations of the several membrane bound human cytochrome p450. Furthermore, our intension is to use different constitutive promoters in order to express the cytochromes at different levels. 

The next step will be to examine our different cytochrome p450 transformed E.coli bacteria and their ability to impede estrogenic activity and reduce carbamazepine concentration from water. We hope to find and possibly optimize a biological system which can be introduced in bacteria and lead to the removal of PPCP pollutants in water.

Our project has the aim to characterize Cytochrome p450 as a BioBrick and thus add it to the partsregistry of BioBricks, a continuously growing collection of genetic parts that can be mixed and matched to build synthetic biology devices and systems (partsregistry.org). Hence our project contributes to the expansion of the partsregistry, benefitting all researchers with interest in this particular protein, since cytochrome p450 doesn’t exist in the partsregistry. The partsregistry is founded on an open source philosophy, a philosophy we see as a great advantage for development and improvement of the number BioBricks already existing. The addition of cytochrome p450 to this catalogue will perhaps encourage other groups to look at other aspects of the many functions of p450 and thus contribute to the formation of new biological systems – further expanding the field of synthetic biology.