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Literature summary for 5.3.1.5 extracted from

  • Waltman, M.; Yang, Z.; Langan, P.; Graham, D.; Kovalevsky, A.
    Engineering acidic Streptomyces rubiginosus D-xylose isomerase by rational enzyme design (2014), Protein Eng., 27, 59-64.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Streptomyces rubiginosus

Protein Variants

Protein Variants Comment Organism
D287N inactive Streptomyces rubiginosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1 4 D-xylose recombinant His12-tagged wild type enzyme, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
1 1.2 D-xylose mutant enzyme E186Q/N215D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
3
-
D-xylose native wild type enzyme, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
3.4
-
D-xylose mutant enzyme N215D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
3.8
-
D-xylose recombinant His12-tagged wild type enzyme, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
4 5 D-xylose mutant enzyme D57N, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
4.3
-
D-xylose mutant enzyme K289H, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
5
-
D-xylose recombinant His6-tagged wild type enzyme, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
5.3
-
D-xylose mutant enzyme D57N, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
6
-
D-xylose mutant enzyme N247D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
6.5
-
D-xylose mutant enzyme Q256D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
7.4
-
D-xylose mutant enzyme D57H, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
8
-
D-xylose mutant enzyme K289E, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
10
-
D-xylose mutant enzyme N215D, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
15
-
D-xylose mutant enzyme E186Q/N215D, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
27
-
D-xylose mutant enzyme K289H, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
33
-
D-xylose recombinant His6-tagged wild type enzyme, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
37
-
D-xylose mutant enzyme K289E, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
42
-
D-xylose mutant enzyme N247D, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
83
-
D-xylose native wild type enzyme, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
548
-
D-xylose mutant enzyme D57H, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-Xylose Streptomyces rubiginosus
-
D-Xylulose
-
r

Organism

Organism UniProt Comment Textmining
Streptomyces rubiginosus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap crude nickel affinity column chromatography Streptomyces rubiginosus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-Xylose
-
Streptomyces rubiginosus D-Xylulose
-
r

Synonyms

Synonyms Comment Organism
D-xylose isomerase
-
Streptomyces rubiginosus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.007
-
D-xylose mutant enzyme D57H, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.056
-
D-xylose mutant enzyme E186Q/N215D, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.064
-
D-xylose mutant enzyme K289H, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.07
-
D-xylose mutant enzyme K289E, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.074
-
D-xylose mutant enzyme N247D, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.11
-
D-xylose mutant enzyme N247D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.14
-
D-xylose recombinant His12-tagged wild type enzyme, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.15
-
D-xylose mutant enzyme Q256D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.18
-
D-xylose mutant enzyme D57H, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.19
-
D-xylose mutant enzyme N215D, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.31
-
D-xylose recombinant His6-tagged wild type enzyme, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.33
-
D-xylose mutant enzyme D57N, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.41
-
D-xylose native wild type enzyme, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.51
-
D-xylose mutant enzyme K289E, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.66
-
D-xylose mutant enzyme E186Q/N215D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.8
-
D-xylose recombinant His12-tagged wild type enzyme, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.96
-
D-xylose mutant enzyme D57N, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
1.22
-
D-xylose recombinant His6-tagged wild type enzyme, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
1.38
-
D-xylose mutant enzyme K289H, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
2.3
-
D-xylose mutant enzyme N215D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
5.52
-
D-xylose native wild type enzyme, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.00013
-
D-xylose mutant enzyme D57H, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.0018
-
D-xylose mutant enzyme N247D, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.0019
-
D-xylose mutant enzyme K289E, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.0024
-
D-xylose mutant enzyme K289H, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.0037
-
D-xylose mutant enzyme E186Q/N215D, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.0049
-
D-xylose native wild type enzyme, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.007
-
D-xylose mutant enzyme D57N, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.0094
-
D-xylose recombinant His6-tagged wild type enzyme, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.01
-
D-xylose recombinant His12-tagged wild type enzyme, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.018
-
D-xylose mutant enzyme N247D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.019
-
D-xylose mutant enzyme N215D, in 100 mM MES-NaOH (pH 5.8), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.023
-
D-xylose mutant enzyme Q256D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.024
-
D-xylose mutant enzyme D57H, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.059
-
D-xylose mutant enzyme E186Q/N215D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.064
-
D-xylose mutant enzyme K289E, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.181
-
D-xylose mutant enzyme D57N, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.211
-
D-xylose recombinant His12-tagged wild type enzyme, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.24
-
D-xylose recombinant His6-tagged wild type enzyme, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.321
-
D-xylose mutant enzyme K289H, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
0.676
-
D-xylose mutant enzyme N215D, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus
1.84
-
D-xylose native wild type enzyme, in 100 mM HEPES-NaOH (pH 7.7), 20 mM MgCl2, at 25°C Streptomyces rubiginosus