Part:BBa_K3945007

Lanmodulin with Cex cellulose binding domain (LanM-Cex)

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.

A novel fusion protein with lanmodulin fused to the N-terminus of Cellulomonas fimi’s Cex cellulose binding module CBMcex. CBMcex has a strong affinity for a variety of cellulose types, especially microcrystalline cellulose over a wide variety of conditions and pH ranges [2]. Thus, allowing for efficient one-step immobilization and purification of lanmodulin onto cellulose support.

Design

With a flexible and inert Threonine-Proline linker joining the two proteins, the immobilized lanmodulin will be able to operate freely on the surface of the cellulose support. The sequence of both proteins was also codon-optimized for expression in E. coli. A Factor Xa recognition site has been incorporated between the CBM and lanmodulin to cleave the lanmodulin protein free after purification on cellulose

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 88
  
• 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

3.Improved immobilization of fusion proteins via cellulose‐binding domains - Linder - 1998 - Biotechnology and Bioengineering - Wiley Online Library. [accessed 2021 Oct 14]. https://onlinelibrary.wiley.com/doi/pdf/10.1002/(SICI)1097-0290(19981205)60:5%3C642::AID-BIT15%3E3.0.CO;2-8