Team:Heidelberg/Tour
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- | + | Non-Ribosomal Peptide Synthetases are composed of building blocks, which are called modules. Each module shows a distinct specificity for a large variety of different monomers assembling them chain-like to a protein. Therefore, the order of modules determines the sequence of the final proteins. We used the tyrocidine synthetase from <i>Brevibacillus parabrevis</i> to:</p> | |
- | + | <ul> | |
+ | <li style="font-size.14px">employ this modularity to investigate the interchangeability and therefore compatibility of modules</li> | ||
+ | <li style="font-size.14px">engineer custom synthetic peptides <i>in vivo</i> by shuffling modules</li> | ||
+ | <li style="font-size.14px">demonstrate the broad variety of possible products</li> | ||
+ | </ul> | ||
+ | <p style="font-size:14px; text-align:justify"> | ||
+ | Understanding this modularity could provide new insights into combinatorial biology and approaches for creating compound libraries for screening purposes.</p> | ||
<a href="/Team:Heidelberg/Project/Tyrocidine" style="margin-bottom:5%"><button typ="button" class="btn btn-default">Discover more!</button></a> | <a href="/Team:Heidelberg/Project/Tyrocidine" style="margin-bottom:5%"><button typ="button" class="btn btn-default">Discover more!</button></a> | ||
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Revision as of 00:39, 29 October 2013
Take the Tour! Take Your Chance and Join Our Quest!
Overview
We wish to open up the great field of NRPS to the iGEM community.
Therefore, we:
- Proved that Non-Ribosomal Peptide Synthetase modules behave in a modular manner and can be shuffled
- Developed a method for easy Non-Ribosomal Peptide-detection
- Employed a Non-Ribosomal Peptide for the recycling of gold from electronic waste
- Implement a software which enables everyone to design synthetic Non-Ribosomal Peptides
Recycling Gold from Electronic Waste Using the NRP Delftibactin
Due to the fast turn-over of today’s high-tech equipment, millions of tons of electronic waste accumulate each year. It contains tons of gold which is very valuable to the society, not only because of its use in jewelry but also for medical applications. The main approach nowadays is to recycle gold by electrolysis which is highly inefficient and expensive, preventing most of the gold from being recovered. In our project we worked on finding a more efficient and environmentally friendly method to recover this precious metal:
- We managed to recover gold from electronic waste using delftibactin, a NRP produced by Delftia acidovorans
- We managed to clone all genes needed for delftibactin production into E. coli
- We managed to recombinantly express the NRPS responsible for delftibactin production in E. coli
Synthetic Peptides
Non-Ribosomal Peptide Synthetases are composed of building blocks, which are called modules. Each module shows a distinct specificity for a large variety of different monomers assembling them chain-like to a protein. Therefore, the order of modules determines the sequence of the final proteins. We used the tyrocidine synthetase from Brevibacillus parabrevis to:
- employ this modularity to investigate the interchangeability and therefore compatibility of modules
- engineer custom synthetic peptides in vivo by shuffling modules
- demonstrate the broad variety of possible products
Understanding this modularity could provide new insights into combinatorial biology and approaches for creating compound libraries for screening purposes.
Non-Ribosomal Peptide Synthetases are composed of building blocks, which are called modules. Each module shows a distinct specificity for a large variety of different monomers assembling them chain-like to a protein. Therefore, the order of modules determines the sequence of the final proteins. We used the tyrocidine synthetase from Brevibacillus parabrevis to:
- employ this modularity to investigate the interchangeability and therefore compatibility of modules
- engineer custom synthetic peptides in vivo by shuffling modules
- demonstrate the broad variety of possible products
Understanding this modularity could provide new insights into combinatorial biology and approaches for creating compound libraries for screening purposes.
Indigoidine-Tag
Indigoidine is a blue pigment produced by the NRPS module indC. We fused indC with other NRPS modules to yield an assembly line producing peptides labelled with the Indigoidine-Tag. This procedure opens up the possibility for the high-throughput generation of combinatorial NRPS libraries due to its simplicity in assembly, detection, validation and purification. We propose such high-throughput protocols in our RFC100. We optimized the indigoidine synthetase by replacing single domains with both domains from NRPS modules and fully synthetic domains and coexpression with different enzymes required for NRPS activation (see Tag-Optimization).
Software: NRPSDesigner
Features:
- User input: A desired peptide consisting of more than 50 amino acids with different chirality, which are currently present in our database
- The software calculates a NRPS domain sequence, composed of domains from different species, which is capable of producing the desired non-ribosomal peptide
- Via the integrated Software "Gibthon", a former iGEM project, a Gibson cloning strategy is designed
- to make sure our tool can be used beyond this years iGEM competition we furthermore implemented the possibility of adding new domains to our database by entering a DNA sequence
- On the basis of this DNA sequence a domain prediction is carried out, which is based on antismash2
- This de facto enables everyone to design his or her non-ribosomal peptide of interest!
Modeling the Feasibility of Gold Recycling with Delftibactin
In oder to find out whether the use of delftibactin would be feasible for industrial-scale recycling of gold from electronic waste we designed a model for the cost development of such a aproach.
Modeling the Indigoidine Production
This text show only be displayed under model.
Human Practice
The aim of science and synthetic biology in particular is to improve lives by solving problems. We as researchers (-to-be) are therefore working for society. Yet, we can only offer solutions, which have to be approved and applied by the public.
We as iGEM Team Heidelberg have therefore put great effort in communicating with many groups within society to open minds, broaden horizons as well as minimize prejudices and concerns:
- Involving experts
- Engaging the broad public
- Getting inspired by artists
- Intruiging the next generation of scientists
And finally bringing them all together to an open talk evening addressing the question "On the Way to a Synthetic Future?"
Results
- We opened up the usage of NRPS to the iGEM community
- We proved modularity of NRPS on different hierachical layers
- We established a universal and specific tag for the labeling of NRPs
- We submitted a standardized assembly line production procedure for synthetic NRPs: RFC 99 and RFC 100
- We implemented a software which enables everyone to create synthetic NRPS!
- We had a great summer with lots of fun and new experiences!