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Literature summary for 1.1.1.67 extracted from

  • Lucas, J.; Siegel, J.
    Erratum: Quantitative functional characterization of conserved molecular interactions in the active site of mannitol 2-dehydrogenase (2015), Protein Sci., 24, 936-945.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BLR(DE3) cells Pseudomonas fluorescens

Protein Variants

Protein Variants Comment Organism
D230A the mutant shows reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens
E133A the mutant shows reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens
E133Q the mutant shows reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens
H303A the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens
H303A/R373A/K381A the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens
K295A inactive Pseudomonas fluorescens
K381A the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens
N191A the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens
N300A the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens
N300D the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens
N300S the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens
R373A the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme Pseudomonas fluorescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.56
-
D-mannitol mutant enzyme N191A, at pH 7.4 and 25°C Pseudomonas fluorescens
5.98
-
D-mannitol wild type enzyme, at pH 7.4 and 25°C Pseudomonas fluorescens
8.15
-
D-mannitol mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
14.96
-
D-mannitol mutant enzyme E133Q, at pH 7.4 and 25°C Pseudomonas fluorescens
17.49
-
D-mannitol mutant enzyme R373A, at pH 7.4 and 25°C Pseudomonas fluorescens
18.21
-
D-mannitol mutant enzyme E133A, at pH 7.4 and 25°C Pseudomonas fluorescens
20.36
-
D-arabitol mutant enzyme D230A, at pH 7.4 and 25°C Pseudomonas fluorescens
20.42
-
D-mannitol mutant enzyme N300D, at pH 7.4 and 25°C Pseudomonas fluorescens
23.89
-
D-mannitol mutant enzyme K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
33.15
-
D-arabitol mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
42.74
-
D-mannitol mutant enzyme D230A, at pH 7.4 and 25°C Pseudomonas fluorescens
49.95
-
D-mannitol mutant enzyme N300S, at pH 7.4 and 25°C Pseudomonas fluorescens
55.85
-
D-arabitol wild type enzyme, at pH 7.4 and 25°C Pseudomonas fluorescens
68.94
-
D-mannitol mutant enzyme N300A, at pH 7.4 and 25°C Pseudomonas fluorescens
93.85
-
D-mannitol mutant enzyme H303A, at pH 7.4 and 25°C Pseudomonas fluorescens
94.82
-
D-arabitol mutant enzyme N300D, at pH 7.4 and 25°C Pseudomonas fluorescens
125.5
-
D-arabitol mutant enzyme N191A, at pH 7.4 and 25°C Pseudomonas fluorescens
126.5
-
D-arabitol mutant enzyme N300S, at pH 7.4 and 25°C Pseudomonas fluorescens
126.9
-
D-arabitol mutant enzyme E133Q, at pH 7.4 and 25°C Pseudomonas fluorescens
132.1
-
D-arabitol mutant enzyme N300A, at pH 7.4 and 25°C Pseudomonas fluorescens
134.1
-
D-arabitol mutant enzyme E133A, at pH 7.4 and 25°C Pseudomonas fluorescens
144.5
-
D-arabitol mutant enzyme R373A, at pH 7.4 and 25°C Pseudomonas fluorescens
145.1
-
D-arabitol mutant enzyme H303A, at pH 7.4 and 25°C Pseudomonas fluorescens
176.9
-
D-arabitol mutant enzyme K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
1800
-
meso-erythritol Km far above 1800 mM, mutant enzyme E133A, at pH 7.4 and 25°C Pseudomonas fluorescens
1800
-
meso-erythritol Km far above 1800 mM, mutant enzyme E133Q, at pH 7.4 and 25°C Pseudomonas fluorescens
1800
-
meso-erythritol Km far above 1800 mM, mutant enzyme H303A, at pH 7.4 and 25°C Pseudomonas fluorescens
1800
-
meso-erythritol Km far above 1800 mM, mutant enzyme N191A, at pH 7.4 and 25°C Pseudomonas fluorescens
1800
-
meso-erythritol Km far above 1800 mM, mutant enzyme N300A, at pH 7.4 and 25°C Pseudomonas fluorescens
1800
-
meso-erythritol Km far above 1800 mM, mutant enzyme N300S, at pH 7.4 and 25°C Pseudomonas fluorescens
1800
-
meso-erythritol Km far above 1800 mM, wild type enzyme, at pH 7.4 and 25°C Pseudomonas fluorescens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-mannitol + NAD+ Pseudomonas fluorescens
-
D-fructose + NADH + H+
-
r

Organism

Organism UniProt Comment Textmining
Pseudomonas fluorescens O08355
-
-

Purification (Commentary)

Purification (Comment) Organism
HisPur cobalt resin column chromatography Pseudomonas fluorescens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-arabitol + NAD+
-
Pseudomonas fluorescens ?
-
?
D-mannitol + NAD+
-
Pseudomonas fluorescens D-fructose + NADH + H+
-
r
meso-erythritol + NAD+
-
Pseudomonas fluorescens ?
-
?
additional information no activity with glycerol, 1, 2-hexanediol, and 1,2,3-hexanetriol Pseudomonas fluorescens ?
-
?

Synonyms

Synonyms Comment Organism
MDH
-
Pseudomonas fluorescens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.00083
-
D-mannitol mutant enzyme N300D, at pH 7.4 and 25°C Pseudomonas fluorescens
0.001
-
D-mannitol mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.0012
-
meso-erythritol mutant enzyme N191A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.0013
-
D-arabitol mutant enzyme N300D, at pH 7.4 and 25°C Pseudomonas fluorescens
0.0017
-
D-arabitol mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.0024
-
meso-erythritol mutant enzyme N300S, at pH 7.4 and 25°C Pseudomonas fluorescens
0.0029
-
D-arabitol mutant enzyme D230A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.0036
-
meso-erythritol mutant enzyme N300A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.0159
-
meso-erythritol mutant enzyme H303A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.018
-
D-mannitol mutant enzyme R373A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.034
-
D-arabitol mutant enzyme R373A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.044
-
D-arabitol mutant enzyme N191A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.06
-
D-arabitol mutant enzyme N300S, at pH 7.4 and 25°C Pseudomonas fluorescens
0.092
-
D-arabitol mutant enzyme N300A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.096
-
D-mannitol mutant enzyme N300S, at pH 7.4 and 25°C Pseudomonas fluorescens
0.19
-
D-mannitol mutant enzyme K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.209
-
D-mannitol mutant enzyme N191A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.21
-
D-mannitol mutant enzyme N300A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.29
-
D-arabitol mutant enzyme H303A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.37
-
D-arabitol mutant enzyme K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.39
-
D-mannitol mutant enzyme D230A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.489
-
meso-erythritol mutant enzyme E133Q, at pH 7.4 and 25°C Pseudomonas fluorescens
0.49
-
D-mannitol mutant enzyme H303A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.538
-
meso-erythritol mutant enzyme E133A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.599
-
meso-erythritol wild type enzyme, at pH 7.4 and 25°C Pseudomonas fluorescens
3.22
-
D-mannitol mutant enzyme E133Q, at pH 7.4 and 25°C Pseudomonas fluorescens
3.99
-
D-mannitol mutant enzyme E133A, at pH 7.4 and 25°C Pseudomonas fluorescens
4.39
-
D-mannitol wild type enzyme, at pH 7.4 and 25°C Pseudomonas fluorescens
5.5
-
D-arabitol mutant enzyme E133Q, at pH 7.4 and 25°C Pseudomonas fluorescens
6.59
-
D-arabitol wild type enzyme, at pH 7.4 and 25°C Pseudomonas fluorescens
7.3
-
D-arabitol mutant enzyme E133A, at pH 7.4 and 25°C Pseudomonas fluorescens

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Pseudomonas fluorescens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0000021
-
meso-erythritol mutant enzyme N191A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.0000044
-
meso-erythritol mutant enzyme N300S, at pH 7.4 and 25°C Pseudomonas fluorescens
0.0000065
-
meso-erythritol mutant enzyme N300A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.000014
-
D-arabitol mutant enzyme N300D, at pH 7.4 and 25°C Pseudomonas fluorescens
0.000029
-
meso-erythritol mutant enzyme H303A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00004
-
D-mannitol mutant enzyme N300D, at pH 7.4 and 25°C Pseudomonas fluorescens
0.000051
-
D-arabitol mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00014
-
D-arabitol mutant enzyme D230A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00016
-
D-mannitol mutant enzyme H303A/R73A/K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00024
-
D-arabitol mutant enzyme R373A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00035
-
D-arabitol mutant enzyme N191A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00047
-
D-arabitol mutant enzyme N300S, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00069
-
D-arabitol mutant enzyme N300A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00088
-
meso-erythritol mutant enzyme E133Q, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00097
-
meso-erythritol mutant enzyme E133A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00102
-
D-mannitol mutant enzyme R373A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.0018
-
meso-erythritol wild type enzyme, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00192
-
D-mannitol mutant enzyme N300S, at pH 7.4 and 25°C Pseudomonas fluorescens
0.002
-
D-arabitol mutant enzyme H303A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00206
-
D-arabitol mutant enzyme K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00311
-
D-mannitol mutant enzyme N300A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00458
-
D-mannitol mutant enzyme N191A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00522
-
D-mannitol mutant enzyme H303A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00775
-
D-mannitol mutant enzyme K381A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.00908
-
D-mannitol mutant enzyme D230A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.04332
-
D-arabitol mutant enzyme E133Q, at pH 7.4 and 25°C Pseudomonas fluorescens
0.05448
-
D-arabitol mutant enzyme E133A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.118
-
D-arabitol wild type enzyme, at pH 7.4 and 25°C Pseudomonas fluorescens
0.215
-
D-mannitol mutant enzyme E133Q, at pH 7.4 and 25°C Pseudomonas fluorescens
0.219
-
D-mannitol mutant enzyme E133A, at pH 7.4 and 25°C Pseudomonas fluorescens
0.734
-
D-mannitol wild type enzyme, at pH 7.4 and 25°C Pseudomonas fluorescens