Difference between revisions of "Team:NRP-UEA-Norwich"
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<h2 class="title">RESULTS</h2> | <h2 class="title">RESULTS</h2> | ||
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| − | <p class="space20"> | + | <p class="space20">As of 18th September 2015 we have assembled parts and developed methods for testing enzymes that might make enzymatic modifications to glycogen in bacteria and to starch in plants. We are able to detect changes in branching but have not yet detected addition of new groups. This might be below the detection limit of our current assays – we are currently considering different methods for measuring acylation.</p> |
| + | |||
| + | <p class="space20">In plants we have been able to direct our putative modification enzymes to the chloroplast, the site of starch synthesis, and also to show that the quantity of starch produced is not affected. We can now progress to stable transformation of plants to allow us to obtain sufficient quantities of modified starch for analysis. We have also cloned the butyrate synthesis pathway, which requires eight coding sequences. At present the induction of the pathway is reducing the growth of the bacteria, we are currently troubleshooting this and testing the individual steps.</p> | ||
<p class="space20"> You can read more by click on the image on the right.</p> | <p class="space20"> You can read more by click on the image on the right.</p> | ||
Revision as of 13:05, 16 September 2015
ENGINEERING NUTRITION TO INCREASE COLONIC BUTYRATE
THE BACKGROUND
Colon cancer is the second most common cause of cancer death in England and Wales, with 30,000 new cases diagnosed every year and a registered cause of death in half that number.
Recent studies have suggested that high dietary intake of resistant starch may reduce colon cancer and inflammatory bowel disease. Resistant starches escape digestion in the small intestine and are fermented by microbiota in the colon. A small proportion of these colonic bacteria produce short chain fatty acids such as butyrate, which can activate apoptosis in colon cancer cells.
Our project is focused on increasing the amount of butyrate in the colon.
You can read more by clicking on the image on the right.
OUR SOLUTION
We are taking two approaches to increase butyrate levels in the colon.
The first approach is to produce modified starches that might be consumed as a prebiotic. We will test the capability of various putative acyltransferases to transfer acyl groups such as butyryl to the alpha-1,4 carbohydrates produced in bacteria and plants. This will involve modelling carbohydrate molecules to find optimal branching and solubility states.
The second approach is to transfer the butyrate biosynthetic pathway into culturable bacterial species with the long-term aim of producing a probiotic.
You can read more by click on the image on the right.
MODELLING AND SOFTWARE
We used glycogen as a model system to gain a better understanding of the parameters involved in carbohydrate structure, branching and modification. We worked with an existing mathematical model to create structural predictions of carbohydrates.
We developed software to model the putative changes in glycogen structure depending on the position of the butyrate group in the glucose molecule, as growth or branching might be disrupted.
We also built 3D models of carbohydrates to show the importance of the branching degree and number of tiers in the final molecule conformation and we 3D printed them!
Finally, we followed a deterministic approach to model the enzyme kinetics of glycogen branching and debranching.
You can read more by clicking on the image on the right.
RESULTS
As of 18th September 2015 we have assembled parts and developed methods for testing enzymes that might make enzymatic modifications to glycogen in bacteria and to starch in plants. We are able to detect changes in branching but have not yet detected addition of new groups. This might be below the detection limit of our current assays – we are currently considering different methods for measuring acylation.
In plants we have been able to direct our putative modification enzymes to the chloroplast, the site of starch synthesis, and also to show that the quantity of starch produced is not affected. We can now progress to stable transformation of plants to allow us to obtain sufficient quantities of modified starch for analysis. We have also cloned the butyrate synthesis pathway, which requires eight coding sequences. At present the induction of the pathway is reducing the growth of the bacteria, we are currently troubleshooting this and testing the individual steps.
You can read more by click on the image on the right.
PARTS
Add in brief overview of parts
You can read more by click on the image on the right.
PRACTICES
Brief overview of practices
You can read more by clicking on the image on the right.
