Difference between revisions of "Part:BBa K4726000"
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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. | + | ===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. | 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 <i>Escherichia coli</i> BL21 (DE3). We thus recommended to use Escherichia coli Rosetta (DE3) pLysS which helps stabilizing pET recombinants encoding proteins. | 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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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
- 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.