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Literature summary for 3.5.4.27 extracted from

  • Breitung, J.; Schmitz, R.A.; Stetter, K.O.; Thauer, R.K.
    N5,N10-Methenyltetrahydromethanopterin cyclohydrolase from the extreme thermophile Methanopyrus kandleri: increase of the catalytic efficiency (kcat/KM) and thermostability in the presence of salts (1991), Arch. Microbiol., 156, 517-524.
No PubMed abstract available
Search for more literature for 3.5.4.27

General Stability

General Stability Organism
addition of K2HPO4 or other salts prevents inactivation, relatively low concentrations of the salt, below 0.1 mM, are required for the stabilization, potassium salts being more effective than ammonium and sodium salts, at concentrations above 0.5 M, which are required for maximal activity the thermostability decreases again Methanopyrus kandleri

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information effect of salts on Km Methanopyrus kandleri
0.1
-
 N5,N10-methenyl-5,6,7,8-tetrahydromethanopterin 1 M K2HPO4 Methanopyrus kandleri
0.14
-
 N5,N10-methenyl-5,6,7,8-tetrahydromethanopterin 0.6 M K2HPO4 Methanopyrus kandleri
0.2
-
 N5,N10-methenyl-5,6,7,8-tetrahydromethanopterin 0.3 M K2HPO4 Methanopyrus kandleri
2
-
 N5,N10-methenyl-5,6,7,8-tetrahydromethanopterin
-
 Methanopyrus kandleri

Metals/Ions

Metals/Ions Comment Organism Structure
(NH4)2SO4 1.5 M, stimulates Methanopyrus kandleri
ammonium sulfate 1.5 M, maximal 100fold stimulation Methanopyrus kandleri
K2HPO4 maximal activation by 1.5 M K2HPO4, 1 M Na2HPO4, or 1 M Na2SO4 Methanopyrus kandleri
K2HPO4 200fold increase of activity at 1.5 M Methanopyrus kandleri
K2SO4 0.8 M stimulates Methanopyrus kandleri
K2SO4 0.8 M, maximal 100fold stimulation Methanopyrus kandleri
KCl 2.5 M stimulates Methanopyrus kandleri
KCl 2.5 M, maximal 100fold stimulation Methanopyrus kandleri
Na2HPO4 maximal activation by 1.5 M K2HPO4, 1 M Na2HPO4 or 1 M Na2SO4 Methanopyrus kandleri
Na2SO4 maximal activation by 1.5 M K2HPO4, 1 M Na2HPO4 or 1 M Na2SO4 Methanopyrus kandleri
NaCl 2.0 M, stimulates Methanopyrus kandleri
NaCl 2 M, maximal 100fold stimulation Methanopyrus kandleri
NH4Cl optimal concentration is 0.3 M Methanopyrus kandleri
Sodium phosphate 1 M, 200fold stimulation Methanopyrus kandleri

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
41500
-
 1 * 41500, SDS-PAGE Methanopyrus kandleri
42000
-
 non-denaturing PAGE Methanopyrus kandleri

Organism

Organism UniProt Comment Textmining
Methanopyrus kandleri
-
-
-

Oxidation Stability

Oxidation Stability Organism
the purified enzyme is more resistant to inactivation by O2 than the enzyme in cell extract Methanopyrus kandleri

Purification (Commentary)

Purification (Comment) Organism
-
 Methanopyrus kandleri

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5026
-
-
 Methanopyrus kandleri

Storage Stability

Storage Stability Organism
-20°C, under N2, 50 mM Tris/HCl buffer, pH 7, containing 1.3 M KCl, no loss of activity for several months Methanopyrus kandleri
-20°C, under N2, in 50 mM Tris/HCl, pH 7, containing 1.3 M KCl, stable for several months Methanopyrus kandleri
0°C, both under anaeobic and aerobic conditions, the enzyme also loses very little activity within 24 h Methanopyrus kandleri
0°C, both under anaerobic and anaerobic conditions, stable for 24 h Methanopyrus kandleri

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,10-methenyl-5,6,7,8-tetrahydromethanopterin + H2O
-
 Methanopyrus kandleri N5-formyl-5,6,7,8-tetrahydromethanopterin
-
 r

Subunits

Subunits Comment Organism
monomer 1 * 41500, SDS-PAGE Methanopyrus kandleri

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
95
-
-
 Methanopyrus kandleri

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
90
-
 thermostable up to 90°C at 0.04 M K2HPO4 Methanopyrus kandleri
90
-
 60 min, 50 mM tricine/KOH, pH 8, stable Methanopyrus kandleri

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
 Methanopyrus kandleri