Team:Copenhagen/Project/Fungus

The source of CyperMans

fungus destroying power


Cytochrome p450 CYP 79
The Project:


The main goal is to standardize different plant CYP79s (A1, A2 and B1) and deliver them as BioBricks to iGEM. Introduktion

Indtroduction
The plant cytochrome p450 79 are the crucial starting point in the conversion of aminoacids into cyanogenic glycosides important for defense mechanisms in plants. In this biosynthetic pathway the CYP79s and the CYP71 subfamilies are part of a enzyme complex – a so-called metabolon. It has been proposed in the article Dynamic Metabolons [5] that the assembly and disassembly of the metabolons, may differentiate plant defense responses to insect attack and fungal infection.



When isolated, the CYP79 subfamily will produce an intermediate in the biosynthetic pathway, a substance called an oxime. This substance is generally toxic to fungi and is frequently used as chemical fungicides. They function by inhibiting mitochondrial oxidases and thereby promote lipid peroxidation and the production of toxic reactive oxygen species[5]. We will investigate following CYP79s: A1, A2 and B1.The CYP79A1 uses L-Tyrosine as a substrate, which it hydroxylates twice on the amino-group in the presence of NADPH and O2. The products of these reactions are quite unstable and dehydration and decarboxylation will then produce 4-hydroxyphenyl-acetaldehyde oxime. [6] The CYP79B1 isoform is specific for tryptophan and therefore it produces indole 3-acetaldoxime instead, while A2 uses phenylalanine and produces phenylacetaldoxime.

Experimental design:

Ecological and economical prospects:

Our future aim with our project is to determine whether it would be a viable solution to use an oxime producing agent as a type of fungicide. The best scenario would be if the plants themselves was able to produce the oximes and thus protect themselves and eliminating the need for fungicides.

Another use of CyperMan could be in wood protection. Oximes are already used in wood preservatives and paints to protect the wood from fungus and algae. The oximes in use are butanaldoximes, but the oximes produced by CyperMan should have the same effects. By having the cyp under a inducible promoter, induced by a cheap and accesseble compound such as salt water, we could provide a cheap and easy way to protect wood without having to use chemical paint. You could use the bacteria for oxime production and spray it on the wood, where it would kill fungus and algae.

References
[5]Moller, B. L. (2010) Plant science. Dynamic metabolons, Science 330, 1328-1329.
[6]Koch, B. M., Sibbesen, O., Halkier, B. A., Svendsen, I., and Moller, B. L. (1995) The primary sequence of cytochrome P450tyr, the multifunctional N-hydroxylase catalyzing the conversion of L-tyrosine to p-hydroxyphenylacetaldehyde oxime in the biosynthesis of the cyanogenic glucoside dhurrin in Sorghum bicolor (L.) Moench, Arch Biochem Biophys 323, 177-186.