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Literature summary for 5.3.2.3 extracted from

  • Thoden, J.; Vinogradov, E.; Gilbert, M.; Salinger, A.; Holden, H.
    Bacterial sugar 3,4-ketoisomerases structural insight into product stereochemistry (2015), Biochemistry, 54, 4495-4506 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.3.2.3

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta2(DE3) cells Aneurinibacillus thermoaerophilus

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapor diffusion method, using 20% poly(ethylene glycol) 8000, 200 mM NaCl, and 100 mM MOPS (pH 7.0) Aneurinibacillus thermoaerophilus

Protein Variants

Protein Variants Comment Organism
H95R the mutant shows increased activity compared to the wild type enzyme Aneurinibacillus thermoaerophilus
R15Y the mutant shows reduced activity compared to the wild type enzyme Aneurinibacillus thermoaerophilus
R15Y/H95R the mutant shows reduced activity compared to the wild type enzyme Aneurinibacillus thermoaerophilus
Y35F the mutant shows about wild type activity Aneurinibacillus thermoaerophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.045
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme H95R, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
0.15
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme Y35F, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
0.37
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme R15Y/H95R, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
0.41
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme R15Y, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
0.5
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose wild type enzyme, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus

Organism

Organism UniProt Comment Textmining
Aneurinibacillus thermoaerophilus Q6T1W8
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Aneurinibacillus thermoaerophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dTDP-4-dehydro-6-deoxy-alpha-D-glucose
-
 Aneurinibacillus thermoaerophilus dTDP-3-dehydro-6-deoxy-alpha-D-galactose
-
 ?

Synonyms

Synonyms Comment Organism
FdtA
-
 Aneurinibacillus thermoaerophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.18
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme R15Y/H95R, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
0.19
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme R15Y, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
1.23
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme H95R, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
3.3
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme Y35F, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
10.5
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose wild type enzyme, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.46
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme R15Y, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
0.49
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme R15Y/H95R, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
21
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose wild type enzyme, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
22
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme Y35F, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus
27
-
 dTDP-4-dehydro-6-deoxy-alpha-D-glucose mutant enzyme H95R, at pH 7.5 and 37°C Aneurinibacillus thermoaerophilus