Application | Comment | Organism |
---|---|---|
analysis | estimation of the content of L-hydroxyprolines using coupling systems with metabolic enzymes of the trans-4-hydroxy-L-proline pathway (hydroxyproline 2-epimerase (HypE) and cis-4-hydroxy-D-proline dehydrogenase (HypDH)) and the trans-3-hydroxy-L-proline pathway (trans-3-hydroxy-L-proline dehydratase (T3LHypD) and DELTA1-pyrroline-2-carboxylate reductase (Pyr2CR)) from microorganisms. A functional expression system of recombinant HypDH with a heterooligomeric structure is constructed in Escherichia coli cells. Enzymological characterization reveals that the beta-subunit acts as a catalytic subunit, and also that assembly with other subunit(s) improves the kinetics for cis-4-hydroxy-D-proline and thermostability. By using a spectrophotometric assay with different wavelengths, the contents of trans-4-hydroxy-L-proline and trans-3-hydroxy-L-proline are successfully estimated within the ranges of 0.004-1 mM and 0.05-1 mM, respectively, and are consistent with the contents determined by HPLC. This enzymatic method is used to measure the content of trans-4-hydroxy-L-proline in the acid-hydrolysate of collagen, and blood plasma | Colwellia psychrerythraea |
analysis | estimation of the content of L-hydroxyprolines using coupling systems with metabolic enzymes of the trans-4-hydroxy-L-proline pathway (hydroxyproline 2-epimerase (HypE) and cis-4-hydroxy-D-proline dehydrogenase (HypDH)) and the trans-3-hydroxy-L-proline pathway (trans-3-hydroxy-L-proline dehydratase (T3LHypD) and DELTA1-pyrroline-2-carboxylate reductase (Pyr2CR)) from microorganisms. A functional expression system of recombinant HypDH with a heterooligomeric structure is constructed in Escherichia coli cells. Enzymological characterization reveals that the beta-subunit acts as a catalytic subunit, and also that assembly with other subunit(s) improves the kinetics for cis-4-hydroxy-D-proline and thermostability. By using a spectrophotometric assay with different wavelengths, the contents of trans-4-hydroxy-L-proline and trans-3-hydroxy-L-proline are successfully estimated within the ranges of 0.004-1 mM and 0.05-1 mM, respectively, and are consistent with the contents determined by HPLC. This enzymatic method is used to measure the content of trans-4-hydroxy-L-proline in the acid-hydrolysate of collagen, and blood plasma | Azospirillum brasilense |
analysis | estimation of the content of L-hydroxyprolines using coupling systems with metabolic enzymes of the trans-4-hydroxy-L-proline pathway (hydroxyproline 2-epimerase (HypE) and cis-4-hydroxy-D-proline dehydrogenase (HypDH)) and the trans-3-hydroxy-L-proline pathway (trans-3-hydroxy-L-proline dehydratase (T3LHypD) and DELTA1-pyrroline-2-carboxylate reductase (Pyr2CR)) from microorganisms. A functional expression system of recombinant HypDH with a heterooligomeric structure is constructed in Escherichia coli cells. Enzymological characterization reveals that the beta-subunit acts as a catalytic subunit, and also that assembly with other subunit(s) improves the kinetics for cis-4-hydroxy-D-proline and thermostability. By using a spectrophotometric assay with different wavelengths, the contents of trans-4-hydroxy-L-proline and trans-3-hydroxy-L-proline are successfully estimated within the ranges of 0.004-1 mM and 0.05-1 mM, respectively, and are consistent with the contents determined by HPLC. This enzymatic method is used to measure the content of trans-4-hydroxy-L-proline in the acid-hydrolysate of collagen, and blood plasma | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.79 | - |
trans-3-hydroxy-L-proline | pH 8.0, 30°C | Colwellia psychrerythraea | |
9.34 | - |
trans-3-hydroxy-L-proline | pH 8.0, 30°C | Homo sapiens | |
14.3 | - |
trans-3-hydroxy-L-proline | pH 8.0, 30°C | Azospirillum brasilense |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Azospirillum brasilense | V5YXI5 | - |
- |
Colwellia psychrerythraea | Q485S0 | - |
- |
Homo sapiens | Q96EM0 | - |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
7.37 | - |
substrate: trans-3-hydroxy-L-proline, 30°C, pH 8.0 | Homo sapiens |
25.4 | - |
substrate: trans-3-hydroxy-L-proline, 30°C, pH 8.0 | Colwellia psychrerythraea |
26.1 | - |
substrate: trans-3-hydroxy-L-proline, 30°C, pH 8.0 | Azospirillum brasilense |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
trans-3-hydroxy-L-proline | - |
Colwellia psychrerythraea | 1-pyrroline 2-carboxylate + H2O | - |
? | |
trans-3-hydroxy-L-proline | - |
Azospirillum brasilense | 1-pyrroline 2-carboxylate + H2O | - |
? | |
trans-3-hydroxy-L-proline | - |
Homo sapiens | 1-pyrroline 2-carboxylate + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AbLhpH | - |
Azospirillum brasilense |
CpLhpH | - |
Colwellia psychrerythraea |
HsLhpH | - |
Homo sapiens |
T3LHyp dehydratase | - |
Colwellia psychrerythraea |
T3LHyp dehydratase | - |
Azospirillum brasilense |
T3LHyp dehydratase | - |
Homo sapiens |
T3LHypD | - |
Colwellia psychrerythraea |
T3LHypD | - |
Azospirillum brasilense |
T3LHypD | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Colwellia psychrerythraea |
30 | - |
assay at | Azospirillum brasilense |
30 | - |
assay at | Homo sapiens |