Difference between revisions of "Part:BBa K4726000"

 
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<partinfo>BBa_K4726000 short</partinfo>
 
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===Usage and Biology===
 
===Usage and Biology===
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This encodes a N-acetyl-α-D-galactosamine deacetylase from Flavonifractor plautii originating from the human gut microbiota. This enzyme is available on Uniprot with the entry P0DTR4. We added a 6X his-tag on the C-terminus of the protein.
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This enzyme removes the acetyl moiety of N-acetyl-α-D-galactosamine (A antigen). This however does not result in full degradation of A antigen a α-D-galactosamine residue remains. This enzyme works in pair with the α-D-galactosamine galactosaminidase (Part BBa_K4726001) to fully remove the A antigen, producing the H antigen, most commonly known as O type blood.
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The expression of this enzyme led to significant proteotoxicity shown by an impaired growth of <i>Escherichia coli</i> BL21 (DE3). We thus recommended to use Escherichia coli Rosetta (DE3) pLysS which helps stabilizing pET recombinants encoding proteins.
  
 
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'><b>Sequence and Features</b></span>
 
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<partinfo>BBa_K4726000 parameters</partinfo>
 
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===References===
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[1] Rahfeld P, Sim L, Moon H, Constantinescu I, Morgan-Lang C, Hallam SJ, Kizhakkedathu JN, Withers SG. An enzymatic pathway in the human gut microbiome that converts A to universal O type blood. Nat Microbiol. 2019 Sep;4(9):1475-1485. doi: 10.1038/s41564-019-0469-7. Epub 2019 Jun 10. PMID: 31182795.

Latest revision as of 00:35, 10 October 2023


N-acetyl-α-D-galactosamine deacetylase_Flavonifractor plautii

Usage and Biology

This encodes a N-acetyl-α-D-galactosamine deacetylase from Flavonifractor plautii originating from the human gut microbiota. This enzyme is available on Uniprot with the entry P0DTR4. We added a 6X his-tag on the C-terminus of the protein.

This enzyme removes the acetyl moiety of N-acetyl-α-D-galactosamine (A antigen). This however does not result in full degradation of A antigen a α-D-galactosamine residue remains. This enzyme works in pair with the α-D-galactosamine galactosaminidase (Part BBa_K4726001) to fully remove the A antigen, producing the H antigen, most commonly known as O type blood.

The expression of this enzyme led to significant proteotoxicity shown by an impaired growth of Escherichia coli BL21 (DE3). We thus recommended to use Escherichia coli Rosetta (DE3) pLysS which helps stabilizing pET recombinants encoding proteins.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 865
    Illegal EcoRI site found at 1411
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 865
    Illegal EcoRI site found at 1411
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 865
    Illegal EcoRI site found at 1411
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 865
    Illegal EcoRI site found at 1411
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 865
    Illegal EcoRI site found at 1411
    Illegal NgoMIV site found at 1584
    Illegal NgoMIV site found at 1894
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[1] Rahfeld P, Sim L, Moon H, Constantinescu I, Morgan-Lang C, Hallam SJ, Kizhakkedathu JN, Withers SG. An enzymatic pathway in the human gut microbiome that converts A to universal O type blood. Nat Microbiol. 2019 Sep;4(9):1475-1485. doi: 10.1038/s41564-019-0469-7. Epub 2019 Jun 10. PMID: 31182795.