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Studies on the enzymic reduction of amino acids. II. Purification and properties of a D-proline reductase and a proline racemase from Clostridium sticklandii

Stadtman, T.C.; Elliott, P.; J. Biol. Chem. 228, 983-997 (1957)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.21.4.1
NAD+
-
Acetoanaerobium sticklandii
5.1.1.4
2-mercaptoethanol
activates
Acetoanaerobium sticklandii
5.1.1.4
additional information
pyridoxal 5'-phosphate is not involved in racemization
Acetoanaerobium sticklandii
5.1.1.4
SH compounds
activate, e.g. 2-mercaptoethanol, 1,3-dimercaptoethanol
Acetoanaerobium sticklandii
General Stability
EC Number
General Stability
Organism
1.21.4.1
withstands ethanol and acetone fractionation at -20°C, does not lose activity as a result of prolonged dialysis
Acetoanaerobium sticklandii
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.21.4.1
alkaline phosphomonoesterase
preparation from calf intestine, causes extensive inactivation
Acetoanaerobium sticklandii
1.21.4.1
arsenite
-
Acetoanaerobium sticklandii
1.21.4.1
Co2+
-
Acetoanaerobium sticklandii
1.21.4.1
Delta-Aminovalerate
-
Acetoanaerobium sticklandii
1.21.4.1
Fe2+
-
Acetoanaerobium sticklandii
1.21.4.1
human seminal plasma
50% inactivation by incubation at pH 5.5 for 90 min
Acetoanaerobium sticklandii
5.1.1.4
lipoic acid
weak
Acetoanaerobium sticklandii
5.1.1.4
reduced glutathione
weak
Acetoanaerobium sticklandii
5.1.1.4
thioglycolate
weak
Acetoanaerobium sticklandii
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.21.4.1
Ca2+
dialyzed enzyme stimulated by
Acetoanaerobium sticklandii
1.21.4.1
Mg2+
depend on
Acetoanaerobium sticklandii
1.21.4.1
Mn2+
dialyzed enzyme stimulated by
Acetoanaerobium sticklandii
5.1.1.4
Mg2+
37 mM, 25% inhibition
Acetoanaerobium sticklandii
Organic Solvent Stability
EC Number
Organic Solvent
Commentary
Organism
1.21.4.1
Acetone
withstands fractionation at -20°C, does not lose activity as a result of prolonged dialysis
Acetoanaerobium sticklandii
1.21.4.1
Ethanol
withstands fractionation at -20°C, does not lose activity as a result of prolonged dialysis
Acetoanaerobium sticklandii
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.21.4.1
Acetoanaerobium sticklandii
-
formerly referred to as 'an amino acid-fermenting Clostridium, strain HF'
-
1.21.4.1
Clostridium sporogenes
-
-
-
5.1.1.4
Acetoanaerobium sticklandii
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.21.4.1
-
Acetoanaerobium sticklandii
5.1.1.4
-
Acetoanaerobium sticklandii
Reaction
EC Number
Reaction
Commentary
Organism
5.1.1.4
L-proline = D-proline
racemization is accompanied by deuterium incorporation from the solvent into the alpha position of Pro, participation of two equivalent hydrogen acceptor sites
Acetoanaerobium sticklandii
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.21.4.1
20.4
-
-
Acetoanaerobium sticklandii
5.1.1.4
4.8
-
-
Acetoanaerobium sticklandii
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.21.4.1
D-allohydroxyproline + dihydrolipoate
barely detectable activity
2121
Acetoanaerobium sticklandii
5-amino-4-hydroxypentanoic acid + lipoate
-
-
-
ir
1.21.4.1
D-proline + 1,3-dimercaptopropanol
-
2121
Acetoanaerobium sticklandii
5-aminopentanoic acid + ?
-
-
-
ir
1.21.4.1
D-proline + 1,3-dimercaptopropanol
oxidation of 1,3-dimercaptopropanol cannot be replaced by molecular hydrogen, NADH+ or a number of other reducing agents
2121
Acetoanaerobium sticklandii
5-aminopentanoic acid + ?
-
-
-
ir
1.21.4.1
D-proline + 6,8-dimercaptooctanoate
-
2121
Acetoanaerobium sticklandii
5-aminopentanoic acid + ?
-
-
-
ir
1.21.4.1
D-proline + beta-mercaptoethanol
-
2121
Acetoanaerobium sticklandii
?
-
-
-
ir
1.21.4.1
D-proline + dihydrolipoate
-
2121
Acetoanaerobium sticklandii
delta-aminovaleric acid + lipoate
-
-
-
ir
1.21.4.1
D-proline + dihydrolipoate
-
2121
Clostridium sporogenes
delta-aminovaleric acid + lipoate
-
-
-
ir
1.21.4.1
D-proline + dithiol (reduced)
-
2121
Acetoanaerobium sticklandii
5-aminopentanoic acid + oxidized dithiol
-
2121
Acetoanaerobium sticklandii
ir
1.21.4.1
D-proline + DL-lipoic acid
-
2121
Acetoanaerobium sticklandii
5-aminopentanoic acid + ?
-
-
-
ir
5.1.1.4
L-Pro
-
2121
Acetoanaerobium sticklandii
D-Pro
-
2121
Acetoanaerobium sticklandii
-
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
5.1.1.4
24
37
at 37°C the activity is 35-40% greater than at 24°C
Acetoanaerobium sticklandii
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
5.1.1.4
6.6
8.1
pH 5.5: no detectable activity, rapid fall of activity below pH 6.5, activity is relatively stable between pH 6.7 and 8.1
Acetoanaerobium sticklandii
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.21.4.1
NAD+
-
Acetoanaerobium sticklandii
5.1.1.4
2-mercaptoethanol
activates
Acetoanaerobium sticklandii
5.1.1.4
additional information
pyridoxal 5'-phosphate is not involved in racemization
Acetoanaerobium sticklandii
5.1.1.4
SH compounds
activate, e.g. 2-mercaptoethanol, 1,3-dimercaptoethanol
Acetoanaerobium sticklandii
General Stability (protein specific)
EC Number
General Stability
Organism
1.21.4.1
withstands ethanol and acetone fractionation at -20°C, does not lose activity as a result of prolonged dialysis
Acetoanaerobium sticklandii
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.21.4.1
alkaline phosphomonoesterase
preparation from calf intestine, causes extensive inactivation
Acetoanaerobium sticklandii
1.21.4.1
arsenite
-
Acetoanaerobium sticklandii
1.21.4.1
Co2+
-
Acetoanaerobium sticklandii
1.21.4.1
Delta-Aminovalerate
-
Acetoanaerobium sticklandii
1.21.4.1
Fe2+
-
Acetoanaerobium sticklandii
1.21.4.1
human seminal plasma
50% inactivation by incubation at pH 5.5 for 90 min
Acetoanaerobium sticklandii
5.1.1.4
lipoic acid
weak
Acetoanaerobium sticklandii
5.1.1.4
reduced glutathione
weak
Acetoanaerobium sticklandii
5.1.1.4
thioglycolate
weak
Acetoanaerobium sticklandii
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.21.4.1
Ca2+
dialyzed enzyme stimulated by
Acetoanaerobium sticklandii
1.21.4.1
Mg2+
depend on
Acetoanaerobium sticklandii
1.21.4.1
Mn2+
dialyzed enzyme stimulated by
Acetoanaerobium sticklandii
5.1.1.4
Mg2+
37 mM, 25% inhibition
Acetoanaerobium sticklandii
Organic Solvent Stability (protein specific)
EC Number
Organic Solvent
Commentary
Organism
1.21.4.1
Acetone
withstands fractionation at -20°C, does not lose activity as a result of prolonged dialysis
Acetoanaerobium sticklandii
1.21.4.1
Ethanol
withstands fractionation at -20°C, does not lose activity as a result of prolonged dialysis
Acetoanaerobium sticklandii
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.21.4.1
-
Acetoanaerobium sticklandii
5.1.1.4
-
Acetoanaerobium sticklandii
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.21.4.1
20.4
-
-
Acetoanaerobium sticklandii
5.1.1.4
4.8
-
-
Acetoanaerobium sticklandii
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.21.4.1
D-allohydroxyproline + dihydrolipoate
barely detectable activity
2121
Acetoanaerobium sticklandii
5-amino-4-hydroxypentanoic acid + lipoate
-
-
-
ir
1.21.4.1
D-proline + 1,3-dimercaptopropanol
-
2121
Acetoanaerobium sticklandii
5-aminopentanoic acid + ?
-
-
-
ir
1.21.4.1
D-proline + 1,3-dimercaptopropanol
oxidation of 1,3-dimercaptopropanol cannot be replaced by molecular hydrogen, NADH+ or a number of other reducing agents
2121
Acetoanaerobium sticklandii
5-aminopentanoic acid + ?
-
-
-
ir
1.21.4.1
D-proline + 6,8-dimercaptooctanoate
-
2121
Acetoanaerobium sticklandii
5-aminopentanoic acid + ?
-
-
-
ir
1.21.4.1
D-proline + beta-mercaptoethanol
-
2121
Acetoanaerobium sticklandii
?
-
-
-
ir
1.21.4.1
D-proline + dihydrolipoate
-
2121
Acetoanaerobium sticklandii
delta-aminovaleric acid + lipoate
-
-
-
ir
1.21.4.1
D-proline + dihydrolipoate
-
2121
Clostridium sporogenes
delta-aminovaleric acid + lipoate
-
-
-
ir
1.21.4.1
D-proline + dithiol (reduced)
-
2121
Acetoanaerobium sticklandii
5-aminopentanoic acid + oxidized dithiol
-
2121
Acetoanaerobium sticklandii
ir
1.21.4.1
D-proline + DL-lipoic acid
-
2121
Acetoanaerobium sticklandii
5-aminopentanoic acid + ?
-
-
-
ir
5.1.1.4
L-Pro
-
2121
Acetoanaerobium sticklandii
D-Pro
-
2121
Acetoanaerobium sticklandii
-
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
5.1.1.4
24
37
at 37°C the activity is 35-40% greater than at 24°C
Acetoanaerobium sticklandii
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
5.1.1.4
6.6
8.1
pH 5.5: no detectable activity, rapid fall of activity below pH 6.5, activity is relatively stable between pH 6.7 and 8.1
Acetoanaerobium sticklandii