Difference between revisions of "Part:BBa K4759205"

 
 
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<partinfo>BBa_K4759205 short</partinfo>
 
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The application of protein tags,we add a solubilization-promoting tag at the N-end of the P450 enzyme, and in the selected six solubilization-promoting tags (SUMO, GST, MBP, TF, TRX, Nus),
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We add a MBP(solubilization-promoting tag) at the N-terminal of the P450 enzyme
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===Usage and Biology===
 
===Usage and Biology===
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Maltose-binding protein (MBP) is a free protein in the E. coli K12 strain, encoded by the malE gene with a molecular weight of 42 kDa, which is a member of the uptake and breakdown of maltose pathway in E. coli. The most outstanding feature of the MBP label is its strong and highly soluble ability. It is mainly manifested in two aspects: wide range of dissolution and high efficiency. Another important reason for the widespread application of MBP tag is its ability to achieve monopolar purification by specifically binding tags and amylose and eluting under non-denaturing conditions. In addition, fused MBP to either the N terminus or the C terminus was prosoluble. A commercial MBP fusion tag vector is already available to accommodate the research needs.
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We add a solubilization-promoting tag at the N-terminal of the P450 enzyme, and in the selected six solubilization-promoting tags (SUMO, GST, MBP, TF, TRX, Nus), MBP and TF can significantly improve the solubility of Olep protein, but the enzyme activity of Olep is reduced, and the catalytic efficiency is lower than strain without solubilizing tag.
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https://static.igem.wiki/teams/4759/wiki/protein-tags.png
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Fig1: The application of protein tags
  
 
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Latest revision as of 08:20, 12 October 2023


T7-RBS4-MBP-linker-OleP

We add a MBP(solubilization-promoting tag) at the N-terminal of the P450 enzyme


Usage and Biology

Maltose-binding protein (MBP) is a free protein in the E. coli K12 strain, encoded by the malE gene with a molecular weight of 42 kDa, which is a member of the uptake and breakdown of maltose pathway in E. coli. The most outstanding feature of the MBP label is its strong and highly soluble ability. It is mainly manifested in two aspects: wide range of dissolution and high efficiency. Another important reason for the widespread application of MBP tag is its ability to achieve monopolar purification by specifically binding tags and amylose and eluting under non-denaturing conditions. In addition, fused MBP to either the N terminus or the C terminus was prosoluble. A commercial MBP fusion tag vector is already available to accommodate the research needs.

We add a solubilization-promoting tag at the N-terminal of the P450 enzyme, and in the selected six solubilization-promoting tags (SUMO, GST, MBP, TF, TRX, Nus), MBP and TF can significantly improve the solubility of Olep protein, but the enzyme activity of Olep is reduced, and the catalytic efficiency is lower than strain without solubilizing tag.


protein-tags.png

Fig1: The application of protein tags

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 446
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1229
    Illegal AgeI site found at 1327
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 144