Team:Bonn-Rheinbach/Contribution
Project
Contribution
BBa_K4102000
For future iGEM teams, we made the lanmodulin (LanM) protein available as a Basic Part in the registry. BBa_K4102000 functions as a coding part and encodes this protein which has the unique ability to act as a chelator for Rare-Earth Elements. For a complete and detailed characterization see our Part Page. This includes details of our production, purification, and example processes for the application of the protein.
Isolation of REEs
Recombinant LanM can be applied to REE-containing solutions to bind these and potentially separate REEs from any other substance present. Our project has a geological framework and we characterized LanM’s ability by using lanthanide-containing minerals which can be compared to standard REE-Minerals used for mining and refining these elements. Based on its high affinity for REE down to the picomolar range, LanM is a promising chelating agent to extract lanthanides from ore leachates with comparably low REE concentrations. This can also make mining low REE-minerals more attractive.
Purification & Outlook
This basic part was tested by adding the LanM sequence into a pET-19b(+) vector to give us a pET-19b(+)::lanM vector which was then introduced into a bacterial chassis to produce the recombinant protein. IPTG induction and subsequent production of a crude cell extract followed as preparation for the purification step of LanM from the bacteria. On an industrial level, affinity chromatography-free approaches for recombinant protein purification from cell lysates is attractive due to lowering process costs. Both a temperature and pH-based purification was tested and compared due to the useful characteristic of LanM being rather acid and temperature resistant compared to other proteins. Remarkable performance was shown in an acidification-based method. More relevant lab results and details concerning using LanM and how activity can be assessed can be found on our Experience page for our part BBa_K4102000.
Giving as much information as possible while still being transparent about what processes did not work as well should provide the best documentation possible for future iGEM teams also working with LanM or similar proteins that can act as chelators for biomining. Using and testing temperature and pH levels to purify a target protein successfully also shows that making use of unique physicochemical properties of a protein can also lead to good purification levels - something that can be usable for future igem teams operating with exotic proteins.
written by Robin Axt and Friedrich Ehinger