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Team:Heidelberg/Tour
From 2013.igem.org
Project
Notebook
Human Practice
Take the Tour! Take Your Chance and Join Our Quest!
Overview
The iGEM Team Heidelberg introduces NRPS to the iGEM community:
- Proved modular principle of Non-Ribosomal Peptide Synthetase by successful shuffling domains and modules
- Developed a method for easy Non-Ribosomal Peptide-detection
- Employed and tested a concept for the recycling of gold from electronic waste usingNon-Ribosomal Peptides
- Implemented 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
- Creation of fusion NRPS producing peptides labelled with the Indigoidine-Tag
- High-throughput protocols for design, construction and evaluation of combinatorial NRPS libraries (RFC99 and RFC100
- Optimization of the indigoidine synthetase by domain exchanges and coexpression with different PPTases
- Creation of functional synthetic NRPS domains based on multiple sequence alignments
Software: NRPSDesigner
Features:
- Computer aided design of fully synthetic NRPS determined to produce a user-defined short peptide
- Optimal domain assembly based on evolutionary distance
- The curated database stores information of 658 Domains encoded on 99 DNA sequences
- Automated domain recognition for newly entered NRPS sequences
- Integration of Gibthon to facilitate implementation of cloning strategy
- Parts registry interface and SBOL output format
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!
