Team:Imperial College London/M1/PAH

From 2009.igem.org

(Difference between revisions)
m (Phenylketonuria)
(Phenylalanine Hydroxylase Detail)
 
(4 intermediate revisions not shown)
Line 4: Line 4:
[[Image:II09_PAH_Action.png|right|250px]]
[[Image:II09_PAH_Action.png|right|250px]]
-
Phenylketonuria (PKU) is an autosomal recessive genetic disorder where the body is unable to break down the amino acid phenylalanine. This is because of a deficiency of an enzyme called phenylalanine hydroxylase (PAH), which converts phenylalanine to tyrosine. Absence of PAH causes phenylalanine to break down into phenylketone instead. A chronically high level of phenylalanine, called hyperphenylalaninemia will result. Phenylalanine then accumulates in blood and tissues and results in characteristic symptoms of PKU.  Left untreated, PKU can lead to problems with brain development resulting in progressive mental retardation and brain damage.<br><br><br>
+
Phenylketonuria (PKU) is an autosomal recessive genetic disorder where the body is unable to break down the amino acid phenylalanine. This is because of a deficiency of an enzyme called phenylalanine hydroxylase (PAH), which converts phenylalanine to tyrosine. Absence of PAH causes phenylalanine to break down into phenylketone instead. A chronically high level of phenylalanine, called hyperphenylalaninemia will result. Phenylalanine then accumulates in blood and tissues and results in characteristic symptoms of PKU.  Left untreated, PKU can lead to problems with brain development resulting in progressive mental retardation and brain damage.<br><br>
-
[[Image:[[Image:II09_PKU_Testing.jpg|right|300px]]
+
[[Image:II09_PKU_Testing.jpg|left|300px]]
PKU is usually detected at birth, and all new born babies are tested for the disease using a heel prick test. However, there is currently no cure for the condition. Current treatments are centred around following a very strict low-protein diet. By engineering the <i>E.ncapsulator</i> to manufacture PAH, we hope that our cells can be taken as a dietary supplement to replace the missing PAH. This will break down phenylalanine from the diet in the gut, before it is absorbed into the bloodstream.
PKU is usually detected at birth, and all new born babies are tested for the disease using a heel prick test. However, there is currently no cure for the condition. Current treatments are centred around following a very strict low-protein diet. By engineering the <i>E.ncapsulator</i> to manufacture PAH, we hope that our cells can be taken as a dietary supplement to replace the missing PAH. This will break down phenylalanine from the diet in the gut, before it is absorbed into the bloodstream.
 +
<br><br><br>
-
= Phenylalanine Hydroxylase Detail =
+
== Phenylalanine Hydroxylase Detail ==
[[Image:II09_PAH_Action.png|right|300px]]
[[Image:II09_PAH_Action.png|right|300px]]
<br>
<br>
Line 17: Line 18:
We have also modified the PAH sequence on two accounts, in order to remove restriction enzyme recognition sites that would otherwise not allow for the biobricking of the enzyme.
We have also modified the PAH sequence on two accounts, in order to remove restriction enzyme recognition sites that would otherwise not allow for the biobricking of the enzyme.
-
 
+
=== How it was modified ===
 +
The PAH enzyme in its normal form contains 2 restriction enzyme sites that make it unsuitable for submitting directly to the BioBrick registry. For this reason we used <b>Site Directed Mutagenesis</b> to modify one amino acid in this sequence, thereby removing the restriction enzyme recognition sites and making the DNA sequence suitable for submission to the registry.<br><br>
 +
We also submitted a seperate DNA part to the registry that has been modified to make
=== Useful Links ===
=== Useful Links ===
Line 23: Line 26:
[http://www.nspku.org/ NSPKU: National Society for Phenylketonuria]
[http://www.nspku.org/ NSPKU: National Society for Phenylketonuria]
<br><br>
<br><br>
-
<html><center><a href="https://2009.igem.org/Team:Imperial_College_London/M1/EnzymeDelivery#Phenylalanine_Hydroxylase_(PAH)"><img width=150px src="https://static.igem.org/mediawiki/2009/8/8e/II09_Back2Enz.png"></a></center>
+
<html><center><a href="https://2009.igem.org/Team:Imperial_College_London/M1"><img width=150px src="https://static.igem.org/mediawiki/2009/1/10/II09_TourArrow.png"></a>
</html>
</html>
{{Imperial/09/TemplateBottom}}
{{Imperial/09/TemplateBottom}}

Latest revision as of 09:51, 20 October 2009

Contents

Phenylketonuria

II09 PAH Action.png

Phenylketonuria (PKU) is an autosomal recessive genetic disorder where the body is unable to break down the amino acid phenylalanine. This is because of a deficiency of an enzyme called phenylalanine hydroxylase (PAH), which converts phenylalanine to tyrosine. Absence of PAH causes phenylalanine to break down into phenylketone instead. A chronically high level of phenylalanine, called hyperphenylalaninemia will result. Phenylalanine then accumulates in blood and tissues and results in characteristic symptoms of PKU. Left untreated, PKU can lead to problems with brain development resulting in progressive mental retardation and brain damage.

II09 PKU Testing.jpg

PKU is usually detected at birth, and all new born babies are tested for the disease using a heel prick test. However, there is currently no cure for the condition. Current treatments are centred around following a very strict low-protein diet. By engineering the E.ncapsulator to manufacture PAH, we hope that our cells can be taken as a dietary supplement to replace the missing PAH. This will break down phenylalanine from the diet in the gut, before it is absorbed into the bloodstream.


Phenylalanine Hydroxylase Detail

II09 PAH Action.png


Phenylalanine Hydroxylase is an enzyme that catalyses the converstion of phenylalanine to tyrosine. This is conversion is by the addition of a hydroxyl group to the end of the 6-carbon ring of phenylalanine, to form tyrosine.

Since PAH is usually found in the liver, it unsurprisingly lacks any natural resistance to proteases found in the intestine. In order to overcome this problem we introduced a mutation into the structure of PAH to increase its resistance to proteolytic degradation. The delivery of PAH by The Encapsulator is particularly relevant on two accounts. Firstly, it represents a landmark in metabolic subcontraction and secondly, it offers a treatment for a genetic disease.

We have also modified the PAH sequence on two accounts, in order to remove restriction enzyme recognition sites that would otherwise not allow for the biobricking of the enzyme.

How it was modified

The PAH enzyme in its normal form contains 2 restriction enzyme sites that make it unsuitable for submitting directly to the BioBrick registry. For this reason we used Site Directed Mutagenesis to modify one amino acid in this sequence, thereby removing the restriction enzyme recognition sites and making the DNA sequence suitable for submission to the registry.

We also submitted a seperate DNA part to the registry that has been modified to make

Useful Links

[http://www.nspku.org/ NSPKU: National Society for Phenylketonuria]


Mr. Gene   Geneart   Clontech   Giant Microbes