Difference between revisions of "Part:BBa K3945007"
 
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===Usage and Biology===
 
===Usage and Biology===
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<p>Lanmodulin is recently a novel lanthanide binding protein from <I>Methylobacterium extorquens</i> 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.</p>
  
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<p> A novel fusion protein with lanmodulin fused to the N-terminus of <I> Cellulomonas fimi’s </i> 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. </p>
  
 
===Design===
 
===Design===
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<p>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 </p>
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===Experimental Workflow===
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===Sequence and Features===
 
===Sequence and Features===
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===References===
 
===References===
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<p>
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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 </p>
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<p> 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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</p>
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<p> 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 </p>

Latest revision as of 23:25, 21 October 2021

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