Part:BBa_K4726001
α-D-galactosamine galactosaminidase_Flavonifractor plautii
Usage and Biology
This encodes a α-D-galactosamine galactosaminidase from Flavonifractor plautii originating from the human gut microbiota. This enzyme is available on Uniprot under the entry P0DTR5. We added a 6X his-tag at the C-terminus end of the protein.
This enzyme works in pair with P0DTR4 (BBa_K4726000) to fully remove the A antigen. It removes the residual α-D-galactosamine that is left after P0DTR4 takes action. P0DTR5 is not active towards A antigen without prior reaction with P0DTR4. The use of both P0DTR4 and P0DTR5 leads to the production of 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
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 865
Illegal EcoRI site found at 1411 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 865
Illegal EcoRI site found at 1411 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 865
Illegal EcoRI site found at 1411 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 865
Illegal EcoRI site found at 1411 - 25INCOMPATIBLE 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 - 1000COMPATIBLE 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.
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