Difference between revisions of "Part:BBa K5136006"

Revision as of 12:57, 30 September 2024

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

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal XbaI site found at 150
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 831
Illegal XhoI site found at 1231
23
INCOMPATIBLE WITH RFC[23]
Illegal XbaI site found at 150
25
INCOMPATIBLE WITH RFC[25]
Illegal XbaI site found at 150
Illegal AgeI site found at 691
1000
COMPATIBLE WITH RFC[1000]

@@ Line 4: / Line 4: @@
 ===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…