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Literature summary extracted from

  • Grahame, D.A.; Stadtman, T.C.
    Carbon monoxide dehydrogenase from Methanosarcina barkeri. Disaggregation, purification, and physicochemical properties of the enzyme (1987), J. Biol. Chem., 262, 3706-3712.
    View publication on PubMed

General Stability

EC Number General Stability Organism
1.2.7.4 CO renders the enzyme more susceptible to temperature inactivation Methanosarcina barkeri

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.2.7.4 2,3-Butanedione
-
Methanosarcina barkeri
1.2.7.4 CN-
-
Methanosarcina barkeri
1.2.7.4 Glyoxaldehyde inactivation requires enzymatic turnover; only with CO and methyl viologen as substrates Methanosarcina barkeri
1.2.7.4 KCN
-
Methanosarcina barkeri
1.2.7.4 additional information dimethylglyoxime is no inhibitor, formaldehyde and acetaldehyde does not inactivate the enzyme; not inhibited by 20 mM formaldehyde or acetaldehyde Methanosarcina barkeri
1.2.7.4 O2
-
Methanosarcina barkeri

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.7.4 0.5
-
2,3,5-triphenyltetrazolium chloride below 0.5 mM Methanosarcina barkeri
1.2.7.4 0.5
-
2,3,5-triphenyltetrazolium chloride pH 7.0, 25°C Methanosarcina barkeri
1.2.7.4 7.1
-
methyl viologen
-
Methanosarcina barkeri
1.2.7.4 7.1
-
methyl viologen pH 7.0, 25°C Methanosarcina barkeri

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.2.7.4 Fe2+ tightly bound by the enzyme Methanosarcina barkeri
1.2.7.4 Iron contains 15.6 mol of iron per mol of alpha2beta oligomer Methanosarcina barkeri
1.2.7.4 additional information no stimulation by MgSO4, CoCl2, NiCl2 and ZnSO4 Methanosarcina barkeri
1.2.7.4 Ni2+ tightly bound by the enzyme Methanosarcina barkeri
1.2.7.4 Nickel contains 1.3 mol of nickel per mol of alpha2beta2 oligomer Methanosarcina barkeri

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.2.7.4 19700
-
2 * 19700 + 2 * 84500, alpha2beta2, SDS-PAGE Methanosarcina barkeri
1.2.7.4 19700
-
2 * 84500 + 2 * 19700, alpha2beta2, SDS-PAGE Methanosarcina barkeri
1.2.7.4 84500
-
2 * 19700 + 2 * 84500, alpha2beta2, SDS-PAGE Methanosarcina barkeri
1.2.7.4 84500
-
2 * 84500 + 2 * 19700, alpha2beta2, SDS-PAGE Methanosarcina barkeri
1.2.7.4 161000
-
gel filtration Methanosarcina barkeri
1.2.7.4 161000
-
dialyzed enzyme, gel filtration Methanosarcina barkeri
1.2.7.4 205000
-
gradient gel electrophoresis Methanosarcina barkeri
1.2.7.4 3000000
-
high molecular weight form exists under conditions of high ionic strength, gel filtration Methanosarcina barkeri

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.2.7.4 CO + H2O + oxidized ferredoxin Methanosarcina barkeri may participate in methanogenesis by cleavage of acetate, reverse of the reaction in acetate biosynthesis CO2 + reduced ferredoxin
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.2.7.4 Methanosarcina barkeri
-
-
-

Oxidation Stability

EC Number Oxidation Stability Organism
1.2.7.4 extremely sensitive to air, most of the enzyme activity is lost upon exposure to air for 1 min, longer incubation results in complete inactivation Methanosarcina barkeri
1.2.7.4 strong inhibition by oxygen, extremely sensitive, exposed to air activity is lost in less than 1 min Methanosarcina barkeri

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.7.4
-
Methanosarcina barkeri

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.2.7.4 culture condition:acetate-grown cell
-
Methanosarcina barkeri
-
1.2.7.4 culture condition:ammonium malate-grown cell
-
Methanosarcina barkeri
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.2.7.4 133.6
-
-
Methanosarcina barkeri

Storage Stability

EC Number Storage Stability Organism
1.2.7.4 -70°C, samples thawed under anaerobic conditions exhibits full activity even after 6 months of storage Methanosarcina barkeri

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.7.4 CO + H2O + 2,3,5-triphenyltetrazolium chloride
-
Methanosarcina barkeri CO2 + reduced 2,3,5-triphenyltetrazolium chloride
-
?
1.2.7.4 CO + H2O + FAD
-
Methanosarcina barkeri CO2 + FADH2
-
?
1.2.7.4 CO + H2O + ferredoxin
-
Methanosarcina barkeri CO2 + reduced ferredoxin
-
?
1.2.7.4 CO + H2O + FMN
-
Methanosarcina barkeri CO2 + FMNH2
-
?
1.2.7.4 CO + H2O + methyl viologen the enzyme fails to reduce NAD+, NADP+, or 8-hydroxy-5-deazaflavin cofactor 420 Methanosarcina barkeri CO2 + reduced methyl viologen
-
?
1.2.7.4 CO + H2O + oxidized 2,3,5-triphenyltetrazolium chloride
-
Methanosarcina barkeri CO2 + reduced 2,3,5-triphenyltetrazolium chloride
-
?
1.2.7.4 CO + H2O + oxidized ferredoxin
-
Methanosarcina barkeri CO2 + reduced ferredoxin
-
?
1.2.7.4 CO + H2O + oxidized ferredoxin may participate in methanogenesis by cleavage of acetate, reverse of the reaction in acetate biosynthesis Methanosarcina barkeri CO2 + reduced ferredoxin
-
?
1.2.7.4 CO + H2O + oxidized flavodoxin
-
Methanosarcina barkeri CO2 + reduced flavodoxin
-
?
1.2.7.4 CO + H2O + oxidized methyl viologen
-
Methanosarcina barkeri CO2 + reduced methyl viologen
-
?
1.2.7.4 CO + H2O + oxidized phenazine methosulfate
-
Methanosarcina barkeri CO2 + reduced phenazine methosulfate
-
?
1.2.7.4 CO + H2O + phenazine methosulfate
-
Methanosarcina barkeri CO2 + reduced phenazine methosulfate
-
?
1.2.7.4 additional information enzyme fails to reduce NAD+, NADP+ or the 8-hydroxy-5-deazaflavin factor F420 Methanosarcina barkeri ?
-
?

Subunits

EC Number Subunits Comment Organism
1.2.7.4 heterotetramer 2 * 84500 + 2 * 19700, alpha2beta2, SDS-PAGE Methanosarcina barkeri
1.2.7.4 tetramer 2 * 19700 + 2 * 84500, alpha2beta2, SDS-PAGE Methanosarcina barkeri

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.2.7.4 additional information
-
CO renders the enzyme more susceptible to temperature inactivation Methanosarcina barkeri
1.2.7.4 24
-
10 min, about 30% loss of activity in presence of CO, stable in absence of CO Methanosarcina barkeri
1.2.7.4 39
-
10 min, 45% loss of activity in presence of CO, stable in absence of CO Methanosarcina barkeri
1.2.7.4 62
-
10 min, about 65% loss of activity in presence of CO, about 5% loss of activity in absence of CO Methanosarcina barkeri
1.2.7.4 80
-
10 min, about 85% loss of activity in presence of CO, about 20% loss of activity in absence of CO Methanosarcina barkeri
1.2.7.4 80 104 most of the activity remains after heating at 80°C, nearly complete inactivation after heating at 104°C, greater loss of activity at all temperatures when heated in presence of CO Methanosarcina barkeri
1.2.7.4 104
-
10 min, complete loss of activity in presence or in absence of CO Methanosarcina barkeri

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.2.7.4 7 9
-
Methanosarcina barkeri

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.2.7.4 5 11 very little activity outside this range Methanosarcina barkeri