Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Mesorhizobium loti |
Crystallization (Comment) | Organism |
---|---|
homology modeling based in the Rhizobium loti structure | Mesorhizobium loti |
in complex with in complex with Co2+, 2-oxoglutarate as cofactors, and L-proline or L-pipecolic acid. The active site is composed of a distorted jelly roll beta-sheet core, which is sandwiched by the N-terminal and C-terminal alpha-helical domains. Co2+ is coordinated by residues H106, H154, and D108 | Mesorhizobium loti |
Protein Variants | Comment | Organism |
---|---|---|
V95A | no increase in hydroxylation activity nor any improvement in the cis-5/cis-3 product ratio | Mesorhizobium loti |
V95A/V97A | no increase in hydroxylation activity nor any improvement in the cis-5/cis-3 product ratio | Mesorhizobium loti |
V95W | mutation significantly reduces the amount of cis-3-hydroxy-L-pipecolic acid (<5%) production compared with that of cis-5-hydroxy-L-pipecolic acid | Mesorhizobium loti |
V97A | 2.5fold increase in production of cis-5-hydroxy-L-pipecolic acid, ratio of cis-5/cis-3 products is similar to wild-type | Mesorhizobium loti |
V97C | 4fold increase in production of cis-5-hydroxy-L-pipecolic acid, ration of §-cis/5cis products is similar to wild-type | Mesorhizobium loti |
V97F | the cis-5/cis-3 product ratio improves from 1.4 for the wild-type enzyme to 5.3, the total amount of product is similar to wild-type | Mesorhizobium loti |
V97F/V95W | significantly improved regioselectivity toward cis-5-hydroxy-L-pipecolic acid | Mesorhizobium loti |
V97F/V95W/E114G | significantly improved regioselectivity toward cis-5-hydroxy-L-pipecolic acid and high productivity | Mesorhizobium loti |
V97Y | the cis-5/cis-3 product ratio improves from 1.4 for the wild-type enzyme to 9.0, the total amount of product decreases | Mesorhizobium loti |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Iron | - |
Mesorhizobium loti |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mesorhizobium loti | Q92LF6 | - |
- |
Mesorhizobium loti | Q989T9 | - |
- |
Mesorhizobium loti 1021 | Q92LF6 | - |
- |
Mesorhizobium loti MAFF303099 | Q989T9 | - |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.1 | - |
mutant V97F, product cis-3-hydroxy-L-pipecolic acid, pH 7.2, 30°C | Mesorhizobium loti |
0.29 | - |
wild-type, product cis-5-hydroxy-L-pipecolic acid, pH 7.2, 30°C | Mesorhizobium loti |
0.34 | - |
wild-type, product cis-3-hydroxy-L-pipecolic acid, pH 7.2, 30°C | Mesorhizobium loti |
0.79 | - |
mutant V97F, product cis-5-hydroxy-L-pipecolic acid, pH 7.2, 30°C | Mesorhizobium loti |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-pipecolic acid + 2-oxoglutarate + O2 | - |
Mesorhizobium loti | cis-5-hydroxy-L-pipecolic acid + cis-3-hydroxy-L-pipecolic acid + succinate + CO2 | the ratio between cis-5/cis-3 isomers is about 1:7 | ? | |
L-pipecolic acid + 2-oxoglutarate + O2 | - |
Mesorhizobium loti | cis-5-hydroxy-L-pipecolic acid + cis-3-hydroxy-L-pipecolic acid + succinate + CO2 | the ratio between cis-5/cis-3 isomers is about 2:1 | ? | |
L-pipecolic acid + 2-oxoglutarate + O2 | - |
Mesorhizobium loti MAFF303099 | cis-5-hydroxy-L-pipecolic acid + cis-3-hydroxy-L-pipecolic acid + succinate + CO2 | the ratio between cis-5/cis-3 isomers is about 1:7 | ? | |
L-pipecolic acid + 2-oxoglutarate + O2 | - |
Mesorhizobium loti 1021 | cis-5-hydroxy-L-pipecolic acid + cis-3-hydroxy-L-pipecolic acid + succinate + CO2 | the ratio between cis-5/cis-3 isomers is about 2:1 | ? | |
L-proline + 2-oxoglutarate + O2 | - |
Mesorhizobium loti | cis-4-hydroxy-L-proline + succinate + CO2 | - |
? | |
L-proline + 2-oxoglutarate + O2 | - |
Mesorhizobium loti MAFF303099 | cis-4-hydroxy-L-proline + succinate + CO2 | - |
? | |
L-proline + 2-oxoglutarate + O2 | - |
Mesorhizobium loti 1021 | cis-4-hydroxy-L-proline + succinate + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
cis-P4H | - |
Mesorhizobium loti |
mlr6283 | - |
Mesorhizobium loti |
R03107 | - |
Mesorhizobium loti |