Part:BBa_K5136006
CYP199A4 T253D-His tag
Biology
CYP199A4 is a NADH-dependent cytochrome P450 monooxygenase from Rhodopseudomonas palustris cytochrome P450, a heme-dependent enzyme that is a versatile bio-oxidation catalyst for C–X (e.g., X = H, N, S) bond oxidations (1). CYP199A4 can also function as peroxygenase. The engineered CYP199A4 peroxygenases showed good functional group tolerance and preferential O-demethylation at the meta- or para-position, indicating potential O-demethylation of H- and G-type lignin monomers (1).
Usage and design
CYP199A4 can cause alkaline fracture of conjugated side chains of lignin and other colored substances such as azo dyes through nucleophilic reaction, increasing the hydrophilicity of the reaction products, which can be easily removed in the subsequent washing process to achieve the purpose of bleaching (2). It has been pointed out that position T253 of CYP199A4 can affect the coordination environment of the active center. (1) Therefore, we carried out saturation mutagenesis on this site, hoping to screen out enzymes with higher activity. Partial amino acid sequence of CYP199A4 T253D: …AGLDDTVNGI…
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 150
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 831
Illegal XhoI site found at 1231 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 150
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 150
Illegal AgeI site found at 691 - 1000COMPATIBLE WITH RFC[1000]
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