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Literature summary extracted from

  • Bleicher, K.; Winter, J.
    Purification and properties of F420- and NADP(+)-dependent alcohol dehydrogenases of Methanogenium liminatans and Methanobacterium palustre, specific for secondary alcohols (1991), Eur. J. Biochem., 200, 43-51.
    View publication on PubMed

General Stability

EC Number General Stability Organism
1.1.1.184 an ammonium sulfate precipitate (3 M, pH 7) of the enzyme loses its activity completely within a few days Methanobacterium palustre
1.1.1.184 minor loss of activity in 50 mM phosphate buffer is observed during purification and storage at 4°C without additional precautions Methanobacterium palustre
1.1.1.184 no conditions to stabilize the enzyme by freezing at -20°C are found. Addition of 1-10% bovine serum albumin, 20-50% glycerol, 20% 1,2-ethanediol or 0.1% Triton X-100 to the enzyme solution in 40 mM phosphate or 40 mM Tris-HC1 pH 7 before freezing for 1 day or longer has no stabilizing effect Methanobacterium palustre
1.1.98.5 unstable in 40 mM phosphate or Tris-HC1 buffer at 4°C and also in low-ionic-strength buffers in which precipitation occurrs and enzyme activity is lost irreversibly. The enzyme can be stabilized by addition of 200 mM sodium sulfate and 10 mM 2-propanol to the buffer Methanofollis liminatans

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.184 1,10-phenanthroline 5 mM, 17% inhibition Methanobacterium palustre
1.1.1.184 4-hydroxymercuribenzoate 0.025 mM, complete inhibition Methanobacterium palustre
1.1.1.184 4-Methylpyrazole 2 mM, 17% inhibition Methanobacterium palustre
1.1.1.184 iodoacetate 2 mM, complete inhibition Methanobacterium palustre
1.1.1.184 additional information no inhibition by 5 mM 2,2'-dipyridyl, EDTA does not inhibit up to 5 mM Methanobacterium palustre
1.1.98.5 1,10-phenanthroline 5 mM, 87% inhibition Methanofollis liminatans
1.1.98.5 2,2'-dipyridyl 2 mM, 18% inhibition Methanofollis liminatans
1.1.98.5 4-hydroxymercuribenzoate 0.1 mM, complete inhibition Methanofollis liminatans
1.1.98.5 KCl
-
 Methanofollis liminatans
1.1.98.5 additional information no inhibition be EDTA (2-5 mM), iodoacetate (2-5 mM), or 4-hydroxymercurobenzoate (0.01-0.1 mM) Methanofollis liminatans
1.1.98.5 NaCl
-
 Methanofollis liminatans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.184 0.021
-
 NADP+ pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 0.025
-
 NADPH pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 0.4
-
 propanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 2.2
-
 (R)-2-butanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 3
-
 2-propanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 3
-
 diacetyl pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 3.5
-
 2-butanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 4.2
-
 Cyclopentanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 4.5
-
 (S)-2-butanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 4.5
-
 Cyclopentanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 6.5
-
 cyclohexanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 9.5
-
 2-butanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 14
-
 2-Pentanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 16
-
 acetoin pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 18
-
 2,3-Butanediol pH 7.5, 37°C Methanobacterium palustre
1.1.98.5 0.008
-
 reduced coenzyme F420 pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 0.018
-
 oxidized coenzyme F420 pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 0.5
-
 propanone pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 0.8
-
 (S)-2-butanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 1.2
-
 2-butanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 2.01
-
 (R)-2-butanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 2.2
-
 2-propanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 4.3
-
 Cyclopentanone pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 6.1
-
 Cyclopentanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 7.2
-
 2-Pentanol pH 5.8, 37°C Methanofollis liminatans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.184 KCl optimal activity is observed in the presence of 500 mM NaCl or KCl Methanobacterium palustre
1.1.1.184 NaCl optimal activity is observed in the presence of 500 mM NaCl or KCl Methanobacterium palustre
1.1.1.184 Zn2+ the enzyme contains 4-8 mol atoms zinc/mol enzyme Methanobacterium palustre
1.1.98.5 additional information the enzyme contains no zinc ions Methanofollis liminatans

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.184 44000
-
 4 * 44000, SDS-PAGE Methanobacterium palustre
1.1.1.184 175000
-
 gel filtration Methanobacterium palustre
1.1.98.5 39000
-
 4 * 39000, SDS-PAGE Methanofollis liminatans
1.1.98.5 150000
-
 gel filtration Methanofollis liminatans

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.184 Methanobacterium palustre
-
-
-
1.1.1.184 Methanobacterium palustre DSM 3108
-
-
-
1.1.98.5 Methanofollis liminatans
-
-
-
1.1.98.5 Methanofollis liminatans DSM 4140
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.184
-
 Methanobacterium palustre
1.1.98.5
-
 Methanofollis liminatans

Storage Stability

EC Number Storage Stability Organism
1.1.1.184 4°C, pH 7, protein concentration above 1.5 mg/ml, strictly anaerobic conditions in 50 mM phosphate, stable for up to 1 year without significant loss of activity Methanobacterium palustre

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.184 (R)-2-butanol + NADP+
-
 Methanobacterium palustre 2-butanone + NADPH + H+
-
 ?
1.1.1.184 (R)-2-butanol + NADP+
-
 Methanobacterium palustre DSM 3108 2-butanone + NADPH + H+
-
 ?
1.1.1.184 (S)-2-butanol + NADP+
-
 Methanobacterium palustre 2-butanone + NADPH + H+
-
 ?
1.1.1.184 (S)-2-butanol + NADP+
-
 Methanobacterium palustre DSM 3108 2-butanone + NADPH + H+
-
 ?
1.1.1.184 2,3-butanediol + NADP+
-
 Methanobacterium palustre acetoin + NADPH + H+
-
 ?
1.1.1.184 2-butanol + NADP+
-
 Methanobacterium palustre 2-butanone + NADPH + H+
-
 r
1.1.1.184 2-pentanol + NADP+
-
 Methanobacterium palustre 2-pentanone + NADPH + H+
-
 r
1.1.1.184 2-propanol + NADP+
-
 Methanobacterium palustre propanone + NADPH + H+ 2-propanone i.e. acetone r
1.1.1.184 cycloheptanone + NADPH + H+
-
 Methanobacterium palustre cycloheptanol + NADP+
-
 ?
1.1.1.184 cyclohexanol + NADP+
-
 Methanobacterium palustre cyclohexanone + NADPH + H+
-
 r
1.1.1.184 cyclohexanol + NADP+
-
 Methanobacterium palustre DSM 3108 cyclohexanone + NADPH + H+
-
 r
1.1.1.184 cyclopentanol + NADP+
-
 Methanobacterium palustre cyclopentanone + NADPH + H+
-
 r
1.1.1.184 diacetyl + NADPH + H+
-
 Methanobacterium palustre acetoin + NADP+
-
 ?
1.1.1.184 diacetyl + NADPH + H+
-
 Methanobacterium palustre DSM 3108 acetoin + NADP+
-
 ?
1.1.1.184 additional information the enzyme catalyses the oxidation of various secondary and cyclic alcohols to the corresponding ketones and the reverse reaction. No primary alcohols are oxidized. 1,3-Butandiol, glycerol, methanol, ethanol, 1,2-ethandiol, 1-propanol, 1-amino-2-propanol, 1-butanol, 1-pentanol, 1-hexanol and benzylalcohol are not oxidized nor are the respective aldehydes or ketones reduced Methanobacterium palustre ?
-
 ?
1.1.1.184 additional information the enzyme catalyses the oxidation of various secondary and cyclic alcohols to the corresponding ketones and the reverse reaction. No primary alcohols are oxidized. 1,3-Butandiol, glycerol, methanol, ethanol, 1,2-ethandiol, 1-propanol, 1-amino-2-propanol, 1-butanol, 1-pentanol, 1-hexanol and benzylalcohol are not oxidized nor are the respective aldehydes or ketones reduced Methanobacterium palustre DSM 3108 ?
-
 ?
1.1.98.5 (R)-2-butanol + oxidized coenzyme F420
-
 Methanofollis liminatans 2-butanone + reduced coenzyme F420
-
 r
1.1.98.5 (R)-2-butanol + oxidized coenzyme F420
-
 Methanofollis liminatans DSM 4140 2-butanone + reduced coenzyme F420
-
 r
1.1.98.5 (S)-2-butanol + oxidized coenzyme F420
-
 Methanofollis liminatans 2-butanone + reduced coenzyme F420
-
 r
1.1.98.5 (S)-2-butanol + oxidized coenzyme F420
-
 Methanofollis liminatans DSM 4140 2-butanone + reduced coenzyme F420
-
 r
1.1.98.5 2-butanone + reduced coenzyme F420
-
 Methanofollis liminatans 2-butanol + oxidized coenzyme F420
-
 r
1.1.98.5 2-butanone + reduced coenzyme F420
-
 Methanofollis liminatans DSM 4140 2-butanol + oxidized coenzyme F420
-
 r
1.1.98.5 2-pentanol + oxidized coenzyme F420
-
 Methanofollis liminatans 2-pentanone + reduced coenzyme F420
-
 r
1.1.98.5 2-pentanol + oxidized coenzyme F420
-
 Methanofollis liminatans DSM 4140 2-pentanone + reduced coenzyme F420
-
 r
1.1.98.5 2-pentanone + reduced coenzyme F420
-
 Methanofollis liminatans 2-pentanol + oxidized coenzyme F420
-
 r
1.1.98.5 2-propanol + oxidized coenzyme F420
-
 Methanofollis liminatans 2-propanone + reduced coenzyme F420 2-propanone i.e. acetone r
1.1.98.5 cyclopentanol + oxidized coenzyme F420
-
 Methanofollis liminatans cyclopentanone + reduced coenzyme F420
-
 r
1.1.98.5 cyclopentanone + reduced coenzyme F420
-
 Methanofollis liminatans cyclopentanol + oxidized coenzyme F420
-
 r
1.1.98.5 additional information the enzyme catalysed the oxidation of various secondary and cyclic alcohols to the corresponding ketones and the reverse reaction. No primary alcohols are oxidized. 1,3-butandiol, glycerol, methanol, ethanol, 1,2-ethandiol, 1 -propanol, 1-amino-2-propanol, 1-butanol, 1-pentanol, 1-hexanol and benzylalcohol are not oxidized nor are the respective aldehydes or ketones reduced Methanofollis liminatans ?
-
 ?
1.1.98.5 additional information the enzyme catalysed the oxidation of various secondary and cyclic alcohols to the corresponding ketones and the reverse reaction. No primary alcohols are oxidized. 1,3-butandiol, glycerol, methanol, ethanol, 1,2-ethandiol, 1 -propanol, 1-amino-2-propanol, 1-butanol, 1-pentanol, 1-hexanol and benzylalcohol are not oxidized nor are the respective aldehydes or ketones reduced Methanofollis liminatans DSM 4140 ?
-
 ?
1.1.98.5 propanone + reduced coenzyme F420 2-propanone i.e. acetone Methanofollis liminatans 2-propanol + oxidized coenzyme F420
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.184 tetramer 4 * 44000, SDS-PAGE Methanobacterium palustre
1.1.98.5 tetramer 4 * 39000, SDS-PAGE Methanofollis liminatans

Synonyms

EC Number Synonyms Comment Organism
1.1.1.184 NADP+-dependent ADH
-
 Methanobacterium palustre
1.1.98.5 F420-dependent alcohol dehydrogenase
-
 Methanofollis liminatans
1.1.98.5 secondary alcohol:F420 oxidoreductase
-
 Methanofollis liminatans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.184 37
-
 assay at Methanobacterium palustre
1.1.1.184 60
-
-
 Methanobacterium palustre
1.1.98.5 42
-
-
 Methanofollis liminatans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.184 5
-
 Cyclohexanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 5
-
 2-Pentanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 6
-
 Cycloheptanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 18
-
 2-Pentanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 26
-
 2,3-Butanediol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 80
-
 Cyclopentanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 86
-
 2-butanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 86
-
 cyclohexanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 106
-
 Cyclopentanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 123
-
 (S)-2-butanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 129
-
 NADPH pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 129
-
 propanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 149
-
 2-butanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 177
-
 (R)-2-butanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 183
-
 acetoin pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 243
-
 NADP+ pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 243
-
 2-propanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 609
-
 diacetyl pH 7.5, 37°C Methanobacterium palustre
1.1.98.5 1.3
-
 2-Pentanone pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 2.1
-
 2-butanone pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 4.4
-
 Cyclopentanone pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 12.8
-
 Cyclopentanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 23.1
-
 2-Pentanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 38.5
-
 (S)-2-butanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 49
-
 2-butanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 51.3
-
 (R)-2-butanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 85
-
 2-propanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 85
-
 oxidized coenzyme F420 pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 161.5
-
 reduced coenzyme F420 pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 161.5
-
 propanone pH 5.8, 37°C Methanofollis liminatans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.184 8
-
-
 Methanobacterium palustre
1.1.98.5 6
-
-
 Methanofollis liminatans

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.184 1.3
-
 2-Pentanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 1.4
-
 2,3-Butanediol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 9
-
 2-butanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 11
-
 acetoin pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 13
-
 cyclohexanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 19
-
 Cyclopentanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 23
-
 Cyclopentanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 27
-
 (S)-2-butanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 42
-
 2-butanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 80
-
 (R)-2-butanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 81
-
 2-propanol pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 203
-
 diacetyl pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 321
-
 propanone pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 5200
-
 NADPH pH 7.5, 37°C Methanobacterium palustre
1.1.1.184 11600
-
 NADP+ pH 7.5, 37°C Methanobacterium palustre
1.1.98.5 1
-
 Cyclopentanone pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 2.1
-
 Cyclopentanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 3.2
-
 2-Pentanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 26
-
 (R)-2-butanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 38.6
-
 2-propanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 40.8
-
 2-butanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 48
-
 (S)-2-butanol pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 323
-
 2-Pentanone pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 4700
-
 oxidized coenzyme F420 pH 5.8, 37°C Methanofollis liminatans
1.1.98.5 20200
-
 reduced coenzyme F420 pH 5.8, 37°C Methanofollis liminatans