Difference between revisions of "Part:BBa K4726002"
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<partinfo>BBa_K4726002 short</partinfo>
 
<partinfo>BBa_K4726002 short</partinfo>
  
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This encodes a α-galactosidase from <i>Bacteroides fragilis</i>. This enzyme is available on Uniprot with the entry Q5LGZ8. 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 Escherichia coli BL21 (DE3). We thus recommended to use Escherichia coli Rosetta (DE3) pLysS which helps stabilizing pET recombinants encoding proteins.
  
 
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Revision as of 00:53, 10 October 2023


α-galactosidase GH110b_Bacteroides fragilis

This encodes a α-galactosidase from Bacteroides fragilis. This enzyme is available on Uniprot with the entry Q5LGZ8. 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 1213
    Illegal EcoRI site found at 1714
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1213
    Illegal EcoRI site found at 1714
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1213
    Illegal EcoRI site found at 1714
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1213
    Illegal EcoRI site found at 1714
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1213
    Illegal EcoRI site found at 1714
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[1] Liu QP, Yuan H, Bennett EP, Levery SB, Nudelman E, Spence J, Pietz G, Saunders K, White T, Olsson ML, Henrissat B, Sulzenbacher G, Clausen H. Identification of a GH110 subfamily of alpha 1,3-galactosidases: novel enzymes for removal of the alpha 3Gal xenotransplantation antigen. J Biol Chem. 2008 Mar 28;283(13):8545-54. doi: 10.1074/jbc.M709020200. Epub 2008 Jan 28. PMID: 18227066; PMCID: PMC2417185.