Coding

Part:BBa_K3945002

Designed by: Ramin Kahidi   Group: iGEM21_Calgary   (2021-10-20)
Revision as of 03:39, 22 October 2021 by ArshiaMostoufi (Talk | contribs) (→‎Design)

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FC-Mutated Lanmodulin (FC LanM)


Usage and Biology

Lanmodulin is recently a novel lanthanide binding protein from Methylobacterium extorquens that displays more than 100-million fold selectivity for rare earth elements [1]. Such affinity and selectivity have not been observed in any previously studied macromolecule. In addition, lanmodulin is extremely robust, capable of withstanding temperatures as high as 95 °C and pH levels as low as 2.5 [2]. Thus, allowing it to be the perfect molecular tool for us in an efficient rare earth recovery system.

We have mutated the lanmodulin sequence using Family Conservation Algorithm to improve the binding capacity of lanmodulin's fourth binding pocket.

Design

Family conservation mutated lanmodulin was generated using a brute force exploration into mutants that were similar in structure and charge distribution but different in the particular amino acids used in each. This was conducted by comprehensively generating sequences for the fourth pocket composed of amino acids that are within the same family of the wildtype lanmodulin. This identified if the interactions causing the unviable binding pocket were due to amino acids structure, not charge distribution.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 111
    Illegal BamHI site found at 186
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

References

1. JA C, ER F, JA M, JV H, TN L. Lanmodulin: A Highly Selective Lanthanide-Binding Protein from a Lanthanide-Utilizing Bacterium. Journal of the American Chemical Society. 2018 [accessed 2021 Sep 17];140(44):15056–15061. https://pubmed.ncbi.nlm.nih.gov/30351021/. doi:10.1021/JACS.8B09842

2. GJ D, JA M, DM P, DW R, JA C, Y J. Selective and Efficient Biomacromolecular Extraction of Rare-Earth Elements using Lanmodulin. Inorganic chemistry. 2020 [accessed 2021 Sep 17];59(17):11855–11867. https://pubmed.ncbi.nlm.nih.gov/32686425/. doi:10.1021/ACS.INORGCHEM.0C01303

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