Activating Compound | Comment | Organism | Structure |
---|---|---|---|
ascorbate | - |
Homo sapiens |
Cloned (Comment) | Organism |
---|---|
gene TMLH, recombinant overexpression of active and stable TMLH as a His-tagged maltose-binding protein fusion in Escherichia coli strain BL21-AI. Coexpression of the enzyme with chaperones GroES/EL increases the expression level by one order of magnitude | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Homo sapiens | |
0.636 | - |
N6,N6,N6-Trimethyl-L-lysine | recombinant MBP-tagged enzyme, pH 7.0, 37°C | Homo sapiens | |
0.696 | - |
N6,N6,N6-Trimethyl-L-lysine | recombinant detagged enzyme, pH 7.0, 37°C | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | dependent on, required for catalysis | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
N6,N6,N6-trimethyl-L-lysine + 2-oxoglutarate + O2 | Homo sapiens | - |
(3S)-3-hydroxy-N6,N6,N6-trimethyl-L-lysine + succinate + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q9NVH6 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant active and stable His- and MBP-tagged TMLH from Escherichia coli strain BL21-AI by nickel or amylose affinity chromatography. It is possible that the His6-tag at the N-terminus of the fusion protein is masked and cannot be accessed by the Ni-beads. Thus, MBP affinity purification is selected as a first purification step, followed by gel filtration, His6-MBP-tag cleavage by recombinant TEV protease | Homo sapiens |
Storage Stability | Organism |
---|---|
4°C, recombinant enzyme, after storage at 4°C for 4 days, MBP-TMLH retains 96% of its activity, while detagged TMLH shows only 77% activity compared to fresh enzyme preparation | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | determination of the (2S,3S)-3-hydroxy-N6,N6,N6-trimethyl-L-lysine and N6,N6,N6-trimethyl-L-lysine is performed by ultraperformance liquid chromatographytandem mass spectrometry (UPLC/MS/MS) in positive ion electrospray mode | Homo sapiens | ? | - |
? | |
N6,N6,N6-trimethyl-L-lysine + 2-oxoglutarate + O2 | - |
Homo sapiens | (3S)-3-hydroxy-N6,N6,N6-trimethyl-L-lysine + succinate + CO2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 44870, recombinant detagged enzyme, SDS-PAGE | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
TMLH | - |
Homo sapiens |
trimethyllysine hydroxylase | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
metabolism | trimethyllysine hydroxylase (TMLH) catalyses the first step in carnitine biosynthesis, the conversion of N6,N6,N6-trimethyl-L-lysine to 3-hydroxy-N6,N6,N6-trimethyl-L-lysine | Homo sapiens |