Team:Copenhagen/Project/Fungus

The source of CyperMans

fungus destroying power


Cytochrome p450 CYP 79
Background information:

It has been proposed in the article Dynamic Metabolons [5] that the assembly and disassembly of enzymes complexes, so-called metabolons, may differentiate plant defense responses to insect attack and fungal infection. The two CYP79 and CYP71 subfamilies are part of a metabolon complex that shifts between assembly and disassembly, according to interactions with proteins or from specific metabolic demands. The CYP79 subfamily can by itself produce a substance called 4-hydroxyphenyl-acetaldeyde oxime. This substance is generally toxic to fungi and 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]. 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-acetaldeyde oxime. [6] The CYP79B1 isoform is specific for tryptophan and therefore it produces indole 3-acetaldoxime instead, while A2 uses phenylalanine and produces phenylacetaldoxime.



The Project:
Our goal is to express different CYP79’s (more specifically from the subfamilies A1, A2 and B1) in E. Coli, to determine whether they can actually be expressed as working proteins in them. Then our idea is introduce a promoter to the system, which reacts to a specific chemical signal that is sent out by fungi.

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.