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Literature summary for 5.3.1.8 extracted from

  • Yeom, S.J.; Kim, Y.S.; Lim, Y.R.; Jeong, K.W.; Lee, J.Y.; Kim, Y.; Oh, D.K.
    Molecular characterization of a novel thermostable mannose-6-phosphate isomerase from Thermus thermophilus (2011), Biochimie, 93, 1659-1667.
    View publication on PubMed
Search for more literature for 5.3.1.8

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli ER2566 cells Thermus thermophilus

Protein Variants

Protein Variants Comment Organism
D138A the mutant shows 59% activity compared to the wild type enzyme Thermus thermophilus
E132A the mutant shows 1.9% activity compared to the wild type enzyme Thermus thermophilus
E132D the mutant shows 0.4% activity compared to the wild type enzyme Thermus thermophilus
E132K inactive Thermus thermophilus
E132Q inactive Thermus thermophilus
E132W inactive Thermus thermophilus
E67A the mutant shows 2.5% activity compared to the wild type enzyme Thermus thermophilus
E67D the mutant shows 3.7% activity compared to the wild type enzyme Thermus thermophilus
E67K inactive Thermus thermophilus
E67Q the mutant shows 0.8% activity compared to the wild type enzyme Thermus thermophilus
E67W inactive Thermus thermophilus
H122A the mutant shows 2.8% activity compared to the wild type enzyme Thermus thermophilus
H122D inactive Thermus thermophilus
H122K inactive Thermus thermophilus
H122Q the mutant shows 2.0% activity compared to the wild type enzyme Thermus thermophilus
H122W inactive Thermus thermophilus
H50A the mutant shows 2.2% activity compared to the wild type enzyme Thermus thermophilus
H50D inactive Thermus thermophilus
H50K inactive Thermus thermophilus
H50Q the mutant shows 2.0% activity compared to the wild type enzyme Thermus thermophilus
H50W inactive Thermus thermophilus
K37A the mutant shows 21% activity compared to the wild type enzyme Thermus thermophilus
K65A the mutant shows 15% activity compared to the wild type enzyme Thermus thermophilus
L18A the mutant shows 78% activity compared to the wild type enzyme Thermus thermophilus
L39A the mutant shows 84% activity compared to the wild type enzyme Thermus thermophilus
Q48A the mutant shows 2.0% activity compared to the wild type enzyme Thermus thermophilus
Q48D inactive Thermus thermophilus
Q48K inactive Thermus thermophilus
Q48N inactive Thermus thermophilus
Q48W inactive Thermus thermophilus
R11A the mutant shows 260% activity compared to the wild type enzyme Thermus thermophilus
R142A the mutant shows 2.6% activity compared to the wild type enzyme Thermus thermophilus
R142D inactive Thermus thermophilus
R142K inactive Thermus thermophilus
R142N inactive Thermus thermophilus
R142W inactive Thermus thermophilus
W13A the mutant shows 0.4% activity compared to the wild type enzyme Thermus thermophilus
W13D inactive Thermus thermophilus
W13F the mutant shows 41% activity compared to the wild type enzyme Thermus thermophilus
W13H the mutant shows 52% activity compared to the wild type enzyme Thermus thermophilus
W13K inactive Thermus thermophilus
W13Q inactive Thermus thermophilus
W13Y the mutant shows 68% activity compared to the wild type enzyme Thermus thermophilus
W69A the mutant shows 50% activity compared to the wild type enzyme Thermus thermophilus
Y124A the mutant shows 52% activity compared to the wild type enzyme Thermus thermophilus

Inhibitors

Inhibitors Comment Organism Structure
Ba2+ about 30% inhibition at 0.5 mM Thermus thermophilus
EDTA complete inhibition at 0.5 mM Thermus thermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
 D-mannose 6-phosphate mutant enzyme E132A, pH 7.0, 80°C Thermus thermophilus
0.18
-
 D-mannose 6-phosphate mutant enzyme E67A, pH 7.0, 80°C Thermus thermophilus
0.21
-
 D-mannose 6-phosphate wild type enzyme, pH 7.0, 80°C Thermus thermophilus
0.22
-
 D-fructose 6-phosphate wild type enzyme, pH 7.0, 80°C Thermus thermophilus
0.27
-
 D-mannose 6-phosphate mutant enzyme H122A, pH 7.0, 80°C Thermus thermophilus
0.33
-
 D-mannose 6-phosphate mutant enzyme H50A, pH 7.0, 80°C Thermus thermophilus
0.53
-
 D-mannose 6-phosphate mutant enzyme Q48A, pH 7.0, 80°C Thermus thermophilus
0.85
-
 D-mannose 6-phosphate mutant enzyme W13A, pH 7.0, 80°C Thermus thermophilus
1.29
-
 D-mannose 6-phosphate mutant enzyme R142A, pH 7.0, 80°C Thermus thermophilus

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ the enzyme is not stimulated by Ca2+ Thermus thermophilus
Co2+ about 230% activity at 0.5 mM Thermus thermophilus
Cu2+ about 135% activity at 0.5 mM Thermus thermophilus
Fe2+ about 125% activity at 0.5 mM Thermus thermophilus
Mg2+ about 175% activity at 0.5 mM Thermus thermophilus
Mn2+ about 175% activity at 0.5 mM Thermus thermophilus
Zn2+ Zn2+ is present at one molecule per monomer, the enzyme has about 240% activity in the presence of 0.5 mM Zn2+ Thermus thermophilus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
29000
-
 1 * 29000, SDS-PAGE Thermus thermophilus
29054
-
 1 * 29054, calculated from amino acid sequence Thermus thermophilus
29100
-
 gel filtration Thermus thermophilus

Organism

Organism UniProt Comment Textmining
Thermus thermophilus Q5SIM4
-
-
Thermus thermophilus KCCM 40879 Q5SIM4
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap HP column chromatography Thermus thermophilus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.01
-
 wild type enzyme, using ribose 5-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
0.3
-
 mutant enzyme E132D, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
0.3
-
 mutant enzyme W13A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
0.6
-
 mutant enzyme E67Q, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
0.74
-
 wild type enzyme, using ribulose 5-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
1.4
-
 mutant enzyme E132A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
1.5
-
 mutant enzyme H122Q, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
1.5
-
 mutant enzyme H50Q, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
1.5
-
 mutant enzyme Q48A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
1.7
-
 mutant enzyme H50A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
1.9
-
 mutant enzyme E67A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
1.9
-
 mutant enzyme R142A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
2.1
-
 mutant enzyme H122A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
2.8
-
 mutant enzyme E67D, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
3 8 mutant enzyme W69A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
4.5
-
 wild type enzyme, using D-fructose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
11
-
 mutant enzyme K65A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
16
-
 mutant enzyme K37A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
31
-
 mutant enzyme W13F, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
39
-
 mutant enzyme W13H, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
39
-
 mutant enzyme Y124A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
40
-
 mutant enzyme L39A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
44
-
 mutant enzyme D138A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
51
-
 mutant enzyme W13Y, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
58
-
 mutant enzyme L18A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
75
-
 wild type enzyme, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus
194
-
 mutant enzyme R11A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C Thermus thermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-fructose 6-phosphate
-
 Thermus thermophilus D-mannose 6-phosphate
-
 r
D-fructose 6-phosphate
-
 Thermus thermophilus KCCM 40879 D-mannose 6-phosphate
-
 r
D-Mannose 6-phosphate best substrate Thermus thermophilus D-Fructose 6-phosphate
-
 r
D-Mannose 6-phosphate best substrate Thermus thermophilus KCCM 40879 D-Fructose 6-phosphate
-
 r
D-ribose 5-phosphate very low activity Thermus thermophilus D-ribulose 5-phosphate
-
 r
D-ribose 5-phosphate very low activity Thermus thermophilus KCCM 40879 D-ribulose 5-phosphate
-
 r
additional information no activity with D-glucose 6-phosphate and arabinose 5-phosphate Thermus thermophilus ?
-
 ?
additional information no activity with D-glucose 6-phosphate and arabinose 5-phosphate Thermus thermophilus KCCM 40879 ?
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 29000, SDS-PAGE Thermus thermophilus
monomer 1 * 29054, calculated from amino acid sequence Thermus thermophilus

Synonyms

Synonyms Comment Organism
mannose-6-phosphate isomerase
-
 Thermus thermophilus
MPI
-
 Thermus thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
-
 Thermus thermophilus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
70 90
-
 Thermus thermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
64 85 the half-lives of the enzyme at 65, 70, 75, 80, and 85°C are 13, 6.5, 3.7, 1.8, and 0.2 h, respectively Thermus thermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
11
-
 D-mannose 6-phosphate mutant enzyme W13A, pH 7.0, 80°C Thermus thermophilus
19
-
 D-mannose 6-phosphate mutant enzyme E132A, pH 7.0, 80°C Thermus thermophilus
31
-
 D-mannose 6-phosphate mutant enzyme E67A, pH 7.0, 80°C Thermus thermophilus
37
-
 D-mannose 6-phosphate mutant enzyme Q48A, pH 7.0, 80°C Thermus thermophilus
54
-
 D-mannose 6-phosphate mutant enzyme H50A, pH 7.0, 80°C Thermus thermophilus
70
-
 D-mannose 6-phosphate mutant enzyme R142A, pH 7.0, 80°C Thermus thermophilus
75
-
 D-mannose 6-phosphate mutant enzyme H122A, pH 7.0, 80°C Thermus thermophilus
141
-
 D-fructose 6-phosphate wild type enzyme, pH 7.0, 80°C Thermus thermophilus
1371
-
 D-mannose 6-phosphate wild type enzyme, pH 7.0, 80°C Thermus thermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
 Thermus thermophilus

pH Range

pH Minimum pH Maximum Comment Organism
6.5 8.5
-
 Thermus thermophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
13
-
 D-mannose 6-phosphate mutant enzyme W13A, pH 7.0, 80°C Thermus thermophilus
54
-
 D-mannose 6-phosphate mutant enzyme R142A, pH 7.0, 80°C Thermus thermophilus
69
-
 D-mannose 6-phosphate mutant enzyme Q48A, pH 7.0, 80°C Thermus thermophilus
109
-
 D-mannose 6-phosphate mutant enzyme E132A, pH 7.0, 80°C Thermus thermophilus
166
-
 D-mannose 6-phosphate mutant enzyme H50A, pH 7.0, 80°C Thermus thermophilus
174
-
 D-mannose 6-phosphate mutant enzyme E67A, pH 7.0, 80°C Thermus thermophilus
278
-
 D-mannose 6-phosphate mutant enzyme H122A, pH 7.0, 80°C Thermus thermophilus
641
-
 D-fructose 6-phosphate wild type enzyme, pH 7.0, 80°C Thermus thermophilus
6685
-
 D-mannose 6-phosphate wild type enzyme, pH 7.0, 80°C Thermus thermophilus