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Literature summary extracted from

  • Krahulec, S.; Armao, G.C.; Klimacek, M.; Nidetzky, B.
    Enzymes of mannitol metabolism in the human pathogenic fungus Aspergillus fumigatus--kinetic properties of mannitol-1-phosphate 5-dehydrogenase and mannitol 2-dehydrogenase, and their physiological implications (2011), FEBS J., 278, 1264-1276.
    View publication on PubMed

Application

EC Number Application Comment Organism
1.1.1.17 pharmacology inhibition of AfM1PDH might be a useful target for therapy of Aspergillus fumigatus infections Aspergillus fumigatus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.17 recombinant expression in Escherichia coli Aspergillus fumigatus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.17 additional information poor effects by ATP, ADP and AMP Aspergillus fumigatus
1.1.1.17 NADH
-
 Aspergillus fumigatus
1.1.1.67 ADP
-
 Aspergillus fumigatus
1.1.1.67 AMP
-
 Aspergillus fumigatus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.17 additional information
-
 additional information steady-state kinetics and kinetic mechanism, binding of D-mannitol 1-phosphate and NAD+ is random, whereas D-fructose 6-phosphate binds only after NADH has bound to the enzyme. Hydride transfer is rate-determining for D-mannitol 1-phosphate oxidation by AfM1PDH. AfM1PDH behaves kinetically as a fructose 6-phosphate reductase, overview Aspergillus fumigatus
1.1.1.17 0.014
-
 NADH pH 10.0, 25°C, recombinant enzyme Aspergillus fumigatus
1.1.1.17 0.13
-
 D-mannitol 1-phosphate pH 7.1, 25°C, recombinant enzyme Aspergillus fumigatus
1.1.1.17 0.8
-
 NAD+ pH 7.1, 25°C, recombinant enzyme Aspergillus fumigatus
1.1.1.17 3.2
-
 D-fructose 6-phosphate pH 10.0, 25°C, recombinant enzyme Aspergillus fumigatus
1.1.1.67 0.015
-
 NADH pH 7.1, 25°C Aspergillus fumigatus
1.1.1.67 0.11
-
 NAD+ pH 7.1, 25°C Aspergillus fumigatus
1.1.1.67 0.6
-
 D-fructose pH 7.1, 25°C Aspergillus fumigatus
1.1.1.67 1.7
-
 D-xylulose pH 7.1, 25°C Aspergillus fumigatus
1.1.1.67 13
-
 D-mannitol pH 10.0, 25°C Aspergillus fumigatus
1.1.1.67 163
-
 D-arabinitol pH 10.0, 25°C Aspergillus fumigatus
1.1.1.67 680
-
 L-sorbitol pH 10.0, 25°C Aspergillus fumigatus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.17 D-mannitol 1-phosphate + NAD+ Aspergillus fumigatus AfM1PDH primarily functions as a D-fructose-6-phosphate reductase and is specific for its natural pair of substrates D-fructose 6-phosphate + NADH + H+
-
 r
1.1.1.67 D-fructose + NADH + H+ Aspergillus fumigatus
-
 D-mannitol + NAD+
-
 r
1.1.1.67 D-mannitol + NAD+ Aspergillus fumigatus free energy profiles for the enzymatic reaction suggest that enzyme primarily acts in D-mannitol oxidation D-fructose + NADH + H+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.17 Aspergillus fumigatus
-
-
-
1.1.1.67 Aspergillus fumigatus Q4WQY4
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.17 D-mannitol 1-phosphate + NAD+ = D-fructose 6-phosphate + NADH + H+ the phosphate moiety in Man-ol1P and Fru6P is essential for substrate recognition and/or catalysis by Aspergillus fumigatus M1PDH. Binding of D-mannitol 1-phosphate and NAD+ is random, whereas D-fructose 6-phosphate binds only after NADH has bound to the enzyme. Hydride transfer is rate-determining for D-mannitol 1-phosphate oxidation by AfM1PDH. The enzyme behaves kinetically as a fructose 6-phosphate reductase Aspergillus fumigatus
a
1.1.1.67 D-mannitol + NAD+ = D-fructose + NADH + H+ binding of substrate and NAD(H) is random for both D-mannitol oxidation and D-fructose reduction. Hydride transfer is rate-determining for D-fructose reduction. Product release steps control the maximum rates in the other direction of the enzymatic reaction Aspergillus fumigatus
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.17 D-mannitol 1-phosphate + NAD+ AfM1PDH primarily functions as a D-fructose-6-phosphate reductase and is specific for its natural pair of substrates Aspergillus fumigatus D-fructose 6-phosphate + NADH + H+
-
 r
1.1.1.17 additional information M1PDH does not catalyze the oxidation of D-mannitol, D-sorbitol, D-ribitol, xylitol, D-xylose, L-xylose, D-glucose, D-mannose, L-arabinose, D-arabinose, D-galactose, L-fucose, and D-lyxose. The enzyme is also inactive above a level of 1% activity with D-fructose 6-phosphate for reduction of D-fructose, L-sorbose, D-xylulose, D-fructose 1,6-bisphosphate, D-glucose 6-phosphate, and D-glucose 1-phosphate Aspergillus fumigatus ?
-
 ?
1.1.1.67 D-arabinitol + NAD+
-
 Aspergillus fumigatus D-xylulose + NADH + H+
-
 r
1.1.1.67 D-fructose + NADH + H+
-
 Aspergillus fumigatus D-mannitol + NAD+
-
 r
1.1.1.67 D-mannitol + NAD+ free energy profiles for the enzymatic reaction suggest that enzyme primarily acts in D-mannitol oxidation Aspergillus fumigatus D-fructose + NADH + H+
-
 r
1.1.1.67 D-xylulose + NADH + H+
-
 Aspergillus fumigatus D-arabinitol + NAD+
-
 ?
1.1.1.67 L-sorbitol + NAD+
-
 Aspergillus fumigatus L-sorbose + NADH + H+
-
 r
1.1.1.67 L-sorbose + NADH + H+
-
 Aspergillus fumigatus L-sorbitol + NAD+
-
 r
1.1.1.67 additional information a D-arabo configuration is required for a polyol substrate to become reactive. The C2 (R) configuration as in mannitol is preferred over the C2 (S) configuration as in D-sorbitol Aspergillus fumigatus ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.17 M1PDH
-
 Aspergillus fumigatus
1.1.1.17 NADH-dependent mannitol-1-phosphate 5-dehydrogenase
-
 Aspergillus fumigatus
1.1.1.67 M2DH
-
 Aspergillus fumigatus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.17 25
-
 assay at Aspergillus fumigatus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.17 40 50 half-life is 20 h at 40°C, the enzyme displays remarkable stability at 40°C and even at 50°C Aspergillus fumigatus
1.1.1.67 25
-
 half-life 3.6 h Aspergillus fumigatus
1.1.1.67 30
-
 half-life 0.42 h Aspergillus fumigatus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.17 10.6
-
 D-mannitol 1-phosphate pH 7.1, 25°C, recombinant enzyme Aspergillus fumigatus
1.1.1.17 132
-
 D-fructose 6-phosphate pH 10.0, 25°C, recombinant enzyme Aspergillus fumigatus
1.1.1.67 60
-
 L-sorbitol pH 10.0, 25°C Aspergillus fumigatus
1.1.1.67 64
-
 D-xylulose pH 7.1, 25°C Aspergillus fumigatus
1.1.1.67 86
-
 D-fructose pH 7.1, 25°C Aspergillus fumigatus
1.1.1.67 162
-
 D-arabinitol pH 10.0, 25°C Aspergillus fumigatus
1.1.1.67 212
-
 D-mannitol pH 10.0, 25°C Aspergillus fumigatus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.17 7.1
-
 assay at, reduction reaction Aspergillus fumigatus
1.1.1.17 10
-
 assay at, oxidation reaction Aspergillus fumigatus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.17 additional information NADP+ is a poor cofactor substrate Aspergillus fumigatus
1.1.1.17 NAD+
-
 Aspergillus fumigatus
1.1.1.17 NADH
-
 Aspergillus fumigatus
1.1.1.67 additional information only trace activity for utilization of NADP+ Aspergillus fumigatus
1.1.1.67 NAD+
-
 Aspergillus fumigatus
1.1.1.67 NADH
-
 Aspergillus fumigatus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.17 0.002
-
 NADH pH 10.0, 25°C, recombinant enzyme Aspergillus fumigatus
1.1.1.67 2.9
-
 AMP pH 7.1, 25°C, reduction of fructose Aspergillus fumigatus
1.1.1.67 4.8
-
 AMP pH 7.1, 25°C, oxidation of mannitol Aspergillus fumigatus
1.1.1.67 5.6
-
 ADP pH 7.1, 25°C, reduction of fructose Aspergillus fumigatus
1.1.1.67 5.7
-
 ADP pH 7.1, 25°C, oxidation of mannitol Aspergillus fumigatus

Expression

EC Number Organism Comment Expression
1.1.1.17 Aspergillus fumigatus M1PDH becomes strongly upregulated during heat shock up

General Information

EC Number General Information Comment Organism
1.1.1.17 additional information ATP, ADP and AMP do not affect the activity of AfM1PDH, suggesting the absence of flux control by cellular energy charge at the level of D-fructose 6-phosphate reduction Aspergillus fumigatus
1.1.1.17 physiological function formation of mannitol is an essential component of the temperature stress response of Aspergillus fumigatus. Enhanced biosynthesis of d-mannitol via AfM1PDH-catalyzed conversion of fructose 6-phosphate might contribute extra robustness to Aspergillus fumigatus under high temperature conditions Aspergillus fumigatus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.17 41
-
 D-fructose 6-phosphate pH 10.0, 25°C, recombinant enzyme Aspergillus fumigatus
1.1.1.17 80
-
 D-mannitol 1-phosphate pH 7.1, 25°C, recombinant enzyme Aspergillus fumigatus
1.1.1.67 0.088
-
 L-sorbitol pH 10.0, 25°C Aspergillus fumigatus
1.1.1.67 0.99
-
 D-arabinitol pH 10.0, 25°C Aspergillus fumigatus
1.1.1.67 1.1
-
 L-sorbose pH 7.1, 25°C Aspergillus fumigatus
1.1.1.67 17
-
 D-mannitol pH 10.0, 25°C Aspergillus fumigatus
1.1.1.67 39
-
 D-xylulose pH 7.1, 25°C Aspergillus fumigatus
1.1.1.67 140
-
 D-fructose pH 7.1, 25°C Aspergillus fumigatus