Literature summary for 4.2.1.77 extracted from

Watanabe, S.; Hiraoka, Y.; Endo, S.; Tanimoto, Y.; Tozawa, Y.; Watanabe, Y.
An enzymatic method to estimate the content of L-hydroxyproline (2015), J. Biotechnol., 199, 9-16 .

Search for more literature for 4.2.1.77

Application

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 [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

Organism	UniProt	Comment	Textmining
Azospirillum brasilense	V5YXI5	-	-
Colwellia psychrerythraea	Q485S0	-	-
Homo sapiens	Q96EM0	-	-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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 [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Colwellia psychrerythraea
30	-	assay at	Azospirillum brasilense
30	-	assay at	Homo sapiens

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Release 2023.1 (January 2023)