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Revision as of 11:31, 15 October 2016

Recovery

Recovery.

Project Achievement
1.

Constructed a multifunctional protein sequences of interest via molecular biological methods, and introduced the constructed plasmids into the engineered bacteria for protein expression. (Learn more)

2.

Searched for methods and the best conditions for the extraction of each protein. (Learn more)

3.

Demonstrated a quick and stable crosslinking process of Triple SpyTag-SUP and Triple SpyTag-mSA with Triple SpyCatcher via covalent bonds. We also optimized this reaction concerning the relevant parameters such as temperature, pH, etc., and confirmed that changes of pH or temperature would not interfere with the whole process. (Learn more)

4.

Demonstrated effective adsorption of uranyl ions by monomeric Triple SpyTag-SUP or protein networks containing the SUP module under a number of conditions. The adsorption was highly efficient and fast, not only under experimental conditions but also in simulated seawater or freshwater containing uranium pollution. (Learn more)

5.

Attached biotin to amino-coated magnetic beads and achieved retrieval of the hydrogel formed via the crosslinking of Triple SpyTag-SUP and Triple SpyTag-mSA with Triple SpyCatcher with a magnet. (Learn more)

6.

Set up a signal peptide library and screened for optimally suited signal peptides in order to efficiently secrete the proteins of interest. We found two signal peptides of high efficiency - those derived from OmpA and LtIIB. (Learn more)

7.

Used all the above-mentioned experiments together to demonstrate that the complete Uranium Reaper system, consisting of Triple SpyTag-SUP, Triple SpyTag-mSA, Triple SpyCatcher and biotin-coated magnetic beads, could effectively handle uranium pollution under simulated real-life conditions in about 2 hours. We aim to optimize this strategy and hope it can be implemented as a uranyl removal kit. (Learn more)

8.

We exchanged the Triple SpyTag-SUP monomer for Triple SpyTag-LBP or Triple SpyTag-CBP, and tried using the same strategy to adsorb lead and cadmium. The results were remarkable, clearly demonstrating that the Uranium Reaper strategy has much potential to be expanded to other heavy metals. (Learn more)

Design


References:

[1] Christopher, D. et al. Streptavidin–biotin technology: improvements and innovations in chemical and biological applications. Appl Microbiol Biotechnol 97, 9343–9353 (2013).

[2] Sau-Ching, W. et al. Engineering monomeric streptavidin and its ligands with infinite affinity in binding but reversibility in interaction. Proteins 77, 404-412 (2009).