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Literature summary for 5.3.1.8 extracted from

  • Lim, Y.R.; Yeom, S.J.; Oh, D.K.
    Production of L-ribose from L-ribulose by a triple-site variant of mannose-6-phosphate isomerase from Geobacillus thermodenitrificans (2012), Appl. Environ. Microbiol., 78, 3880-3884.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli ER2566 cells Geobacillus thermodenitrificans

Protein Variants

Protein Variants Comment Organism
L129F the activity of the mutant is 204% of the activity of the wild type enzyme Geobacillus thermodenitrificans
L129W the activity of the mutant is 10% of the activity of the wild type enzyme Geobacillus thermodenitrificans
L129Y the activity of the mutant is 94% of the activity of the wild type enzyme Geobacillus thermodenitrificans
N90A/L129F the activity of the mutant is about 6fold higher than the activity of the wild type enzyme Geobacillus thermodenitrificans
W17Q/N90A/L129F the specific activity and catalytic efficiency for L-ribulose isomerization of this variant are 3.1 and 7.1fold higher, respectively, than those of the wild type enzyme at pH 7.0 and 70°C in the presence of 1mM Co2+ Geobacillus thermodenitrificans

Inhibitors

Inhibitors Comment Organism Structure
EDTA
-
Geobacillus thermodenitrificans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
8 9 D-mannose 6-phosphate mutant enzyme N90A/L129F, at pH 7.0 and 70°C Geobacillus thermodenitrificans
100
-
D-mannose 6-phosphate mutant enzyme W17Q/N90A/L129F, at pH 7.0 and 70°C Geobacillus thermodenitrificans
149
-
D-mannose 6-phosphate wild type enzyme, at pH 7.0 and 70°C Geobacillus thermodenitrificans
243
-
D-mannose 6-phosphate mutant enzyme L129F, at pH 7.0 and 70°C Geobacillus thermodenitrificans

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ strong stimulation of activity Geobacillus thermodenitrificans
Cu2+ significantly stimulated by Cu2+ Geobacillus thermodenitrificans
Mn2+ significantly stimulated by Mn2+ Geobacillus thermodenitrificans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-Mannose 6-phosphate Geobacillus thermodenitrificans
-
D-Fructose 6-phosphate
-
?

Organism

Organism UniProt Comment Textmining
Geobacillus thermodenitrificans A4ITT1
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-Mannose 6-phosphate
-
Geobacillus thermodenitrificans D-Fructose 6-phosphate
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
23550
-
D-mannose 6-phosphate wild type enzyme, at pH 7.0 and 70°C Geobacillus thermodenitrificans
55120
-
D-mannose 6-phosphate mutant enzyme L129F, at pH 7.0 and 70°C Geobacillus thermodenitrificans
85720
-
D-mannose 6-phosphate mutant enzyme N90A/L129F, at pH 7.0 and 70°C Geobacillus thermodenitrificans
112100
-
D-mannose 6-phosphate mutant enzyme W17Q/N90A/L129F, at pH 7.0 and 70°C Geobacillus thermodenitrificans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
158
-
D-mannose 6-phosphate wild type enzyme, at pH 7.0 and 70°C Geobacillus thermodenitrificans
227
-
D-mannose 6-phosphate mutant enzyme L129F, at pH 7.0 and 70°C Geobacillus thermodenitrificans
965
-
D-mannose 6-phosphate mutant enzyme N90A/L129F, at pH 7.0 and 70°C Geobacillus thermodenitrificans
1120
-
D-mannose 6-phosphate mutant enzyme W17Q/N90A/L129F, at pH 7.0 and 70°C Geobacillus thermodenitrificans