Cloned (Comment) | Organism |
---|---|
expression in Pichia pastoris | Eschscholzia californica |
Protein Variants | Comment | Organism |
---|---|---|
E417Q | solvent isotope effects on kred are equal to 1 for both wild-type and the E417Q mutant, indicating that solvent exchangeable protons are not in flight during or before flavin reduction, thus eliminating a fully concerted mechanism | Eschscholzia californica |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00027 | - |
(S)-reticuline | wild-type, 25°C, pH 9.0 | Eschscholzia californica | |
0.28 | - |
O2 | wild-type, 25°C, pH 9.0 | Eschscholzia californica |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Eschscholzia californica | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-coreximine + O2 | - |
Eschscholzia californica | (13aS)-2,11-dihydroxy-3,10-dimethoxy-5,8,13,13a-tetrahydroisoquinolino[3,2-a]isoquinolin-7-ium + H2O2 | - |
? | |
(S)-laudanosine + H2O2 | - |
Eschscholzia californica | ? + O2 | - |
? | |
(S)-N-methylcoclaurine + O2 | - |
Eschscholzia californica | (S)-coclaurine + H2O2 | - |
? | |
(S)-norsteponine + H2O2 | - |
Eschscholzia californica | ? + O2 | - |
? | |
(S)-reticuline + O2 | - |
Eschscholzia californica | (S)-scoulerine + H2O2 | - |
r | |
(S)-scoulerine + H2O2 | - |
Eschscholzia californica | (S)-reticuline + O2 | - |
r | |
6,7-dimethoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline + H2O2 | - |
Eschscholzia californica | ? + O2 | - |
? | |
additional information | C-H bond cleavage is rate-limiting during flavin reduction. Solvent isotope effects on kred indicate that solvent exchangeable protons are not in flight during or before flavin reduction, thus eliminating a fully concerted mechanism. Deprotonation is not occurring before or during C-H bond cleavage, and a hydroxyl group must be present at C3 for cyclization. This could be due to the methoxy group being less electron-donating than a hydroxyl substituent or to disruption of the interaction between the glutamate and the substrate, causing altered binding. A concerted attack of a methyl amine by the C2-atom and hydride transfer is less likely than attack on a methylene iminium ion intermediate by the C2-atom, indicating that a stepwise mechanism is the likely mechanism | Eschscholzia californica | ? | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.4 | - |
(S)-reticuline | wild-type, 25°C, pH 9.0 | Eschscholzia californica | |
10.5 | - |
O2 | wild-type, 25°C, pH 9.0 | Eschscholzia californica |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
37 | - |
O2 | wild-type, 25°C, pH 9.0 | Eschscholzia californica | |
20000 | - |
(S)-reticuline | wild-type, 25°C, pH 9.0 | Eschscholzia californica |