Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Anisodus acutangulus |
Protein Variants | Comment | Organism |
---|---|---|
K97A | the shows a 3.3fold improved hydroxylation activity compared with the wild-type enzyme | Anisodus acutangulus |
K97E | the shows a 2.7fold improved hydroxylation activity compared with the wild-type enzyme | Anisodus acutangulus |
K97G | the shows a 1.7fold improved hydroxylation activity compared with the wild-type enzyme | Anisodus acutangulus |
S14H | the shows a 2.4fold improved hydroxylation activity compared with the wild-type enzyme | Anisodus acutangulus |
S14P | the shows a 3.7fold improved hydroxylation activity compared with the wild-type enzyme | Anisodus acutangulus |
S14P/K97A | double mutant shows a 4.4fold improved hydroxylation activity compared with the wild-type enzyme, and the in vivo epoxidation activity is also improved by 2.3times. With this mutant, in a 5 l bioreactor with a working volume of 3 l, scopolamine is produced via a single-enzyme-mediated two-step transformation from hyoscyamine of 500 mg/l in 97% yield (1.068 g) | Anisodus acutangulus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0348 | - |
L-hyoscyamine | pH 8.0, 35°C, mutant enzyme S14P/K97A | Anisodus acutangulus | |
0.0416 | - |
L-hyoscyamine | pH 8.0, 35°C | Anisodus acutangulus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-hyoscyamine + 2-oxoglutarate + O2 | Anisodus acutangulus | the enzyme catalyzes the hydroxylation of (-)-hyoscyamine and the subsequent epoxidation of 6beta-hydroxyhyoscyamine to form scopolamine, a natural alkaloid | (6S)-hydroxyhyoscyamine + succinate + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Anisodus acutangulus | A2IBI0 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-hyoscyamine + 2-oxoglutarate + O2 | - |
Anisodus acutangulus | (6S)-hydroxyhyoscyamine + succinate + CO2 | - |
? | |
L-hyoscyamine + 2-oxoglutarate + O2 | the enzyme catalyzes the hydroxylation of (-)-hyoscyamine and the subsequent epoxidation of 6beta-hydroxyhyoscyamine to form scopolamine, a natural alkaloid | Anisodus acutangulus | (6S)-hydroxyhyoscyamine + succinate + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AaH6H | - |
Anisodus acutangulus |
hyoscyamine 6beta-hydroxylase | - |
Anisodus acutangulus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
assay at | Anisodus acutangulus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1 | - |
L-hyoscyamine | pH 8.0, 35°C, wild-type enzyme | Anisodus acutangulus | |
3.9 | - |
L-hyoscyamine | pH 8.0, 35°C, mutant enzyme S14P/K97A | Anisodus acutangulus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Anisodus acutangulus |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme catalyzes the hydroxylation of (-)-hyoscyamine and the subsequent epoxidation of 6beta-hydroxyhyoscyamine to form scopolamine, a natural alkaloid | Anisodus acutangulus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
24.04 | - |
L-hyoscyamine | pH 8.0, 35°C | Anisodus acutangulus | |
112.1 | - |
L-hyoscyamine | pH 8.0, 35°C, mutant enzyme S14P/K97A | Anisodus acutangulus |