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Literature summary for 5.3.1.5 extracted from

  • Patel, D.H.; Cho, E.J.; Kim, H.M.; Choi, I.S.; Bae, H.J.
    Engineering of the catalytic site of xylose isomerase to enhance bioconversion of a non-preferential substrate (2012), Protein Eng. Des. Sel., 25, 331-336.
    View publication on PubMed
Search for more literature for 5.3.1.5

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Thermus thermophilus

Protein Variants

Protein Variants Comment Organism
D254R/D256R complete loss of activity Thermus thermophilus
D256R the mutant shows an increase in the specificity on D-lyxose, L-arabinose and D-mannose Thermus thermophilus

Inhibitors

Inhibitors Comment Organism Structure
Co2+ about 58% residual activity at 1 mM Thermus thermophilus
Cu2+ less than 10% residual activity at 1 mM Thermus thermophilus
Mg2+ about 35% residual activity at 1 mM Thermus thermophilus
Ni2+ less than 5% residual activity at 1 mM Thermus thermophilus
Zn2+ complete inhibition at 1 mM Thermus thermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
138
-
 L-arabinose mutant enzyme D256R, at pH 7.0 and 85°C Thermus thermophilus
185
-
 D-mannose mutant enzyme D256R, at pH 7.0 and 85°C Thermus thermophilus
215
-
 D-Lyxose mutant enzyme D256R, at pH 7.0 and 85°C Thermus thermophilus
605
-
 D-xylose wild type enzyme, at pH 7.0 and 85°C Thermus thermophilus
1005
-
 D-mannose wild type enzyme, at pH 7.0 and 85°C Thermus thermophilus
1450
-
 L-arabinose wild type enzyme, at pH 7.0 and 85°C Thermus thermophilus

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ maximal activity with 1 mM Mn2+ Thermus thermophilus
additional information not stimulated by K+, Ni2+ and Zn2+ Thermus thermophilus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
44000
-
 x * 44000, SDS-PAGE Thermus thermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-Xylose Thermus thermophilus
-
 D-Xylulose
-
 r
D-Xylose Thermus thermophilus HB8 / ATCC 27634 / DSM 579
-
 D-Xylulose
-
 r

Organism

Organism UniProt Comment Textmining
Thermus thermophilus P26997
-
-
Thermus thermophilus HB8 / ATCC 27634 / DSM 579 P26997
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Thermus thermophilus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
132
-
 wild type enzyme, with L-arabinose as substrate, at pH 7.0 and 85°C Thermus thermophilus
273
-
 wild type enzyme, with D-mannose as substrate, at pH 7.0 and 85°C Thermus thermophilus
330
-
 mutant enzyme D256R, with L-arabinose as substrate, at pH 7.0 and 85°C Thermus thermophilus
380
-
 wild type enzyme, with D-lyxose as substrate, at pH 7.0 and 85°C Thermus thermophilus
682
-
 mutant enzyme D256R, with D-mannose as substrate, at pH 7.0 and 85°C Thermus thermophilus
950
-
 mutant enzyme D256R, with D-lyxose as substrate, at pH 7.0 and 85°C Thermus thermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-fructose
-
 Thermus thermophilus D-mannose
-
 r
D-Glucose
-
 Thermus thermophilus D-Fructose
-
 r
D-Glucose
-
 Thermus thermophilus HB8 / ATCC 27634 / DSM 579 D-Fructose
-
 r
D-Lyxose
-
 Thermus thermophilus D-Xylulose
-
 r
D-Lyxose
-
 Thermus thermophilus HB8 / ATCC 27634 / DSM 579 D-Xylulose
-
 r
D-Mannose
-
 Thermus thermophilus D-Fructose
-
 r
D-Mannose
-
 Thermus thermophilus HB8 / ATCC 27634 / DSM 579 D-Fructose
-
 r
D-Xylose
-
 Thermus thermophilus D-Xylulose
-
 r
D-Xylose
-
 Thermus thermophilus HB8 / ATCC 27634 / DSM 579 D-Xylulose
-
 r
L-arabinose
-
 Thermus thermophilus L-ribulose
-
 r
L-arabinose
-
 Thermus thermophilus HB8 / ATCC 27634 / DSM 579 L-ribulose
-
 r
L-ribulose
-
 Thermus thermophilus L-ribose
-
 r

Subunits

Subunits Comment Organism
? x * 44000, SDS-PAGE Thermus thermophilus

Synonyms

Synonyms Comment Organism
D-xylose: ketol-isomerase
-
 Thermus thermophilus
XYLA
-
 Thermus thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
85
-
-
 Thermus thermophilus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
55 95 about 52% activity at 55°C, about 65% activity at 60°C, about 75% activity at 65°C, about 78% activity at 70°C, about 85% activity at 75°C, about 95% activity at 80°C, about 78% activity at 90°C, and about 52% activity at 95°C Thermus thermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2 8 D-mannose mutant enzyme D256R, at pH 7.0 and 85°C Thermus thermophilus
6.5
-
 L-arabinose wild type enzyme, at pH 7.0 and 85°C Thermus thermophilus
13
-
 L-arabinose mutant enzyme D256R, at pH 7.0 and 85°C Thermus thermophilus
18
-
 D-mannose wild type enzyme, at pH 7.0 and 85°C Thermus thermophilus
25
-
 D-Lyxose wild type enzyme, at pH 7.0 and 85°C Thermus thermophilus
38
-
 D-Lyxose mutant enzyme D256R, at pH 7.0 and 85°C Thermus thermophilus

pH Optimum

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

pH Range

pH Minimum pH Maximum Comment Organism
5 9 about 40% activity at pH 5.0, about 60% activity at pH 5.5, about 88% activity at pH 6.0 and 8.0, about 92% activity at pH 6.5 and 7.5, about 80% activity at pH 8.5, about 75% activity at pH 9.0 Thermus thermophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.008
-
 L-arabinose wild type enzyme, at pH 7.0 and 85°C Thermus thermophilus
0.017
-
 D-mannose wild type enzyme, at pH 7.0 and 85°C Thermus thermophilus
0.041
-
 D-Lyxose wild type enzyme, at pH 7.0 and 85°C Thermus thermophilus
0.09
-
 L-arabinose mutant enzyme D256R, at pH 7.0 and 85°C Thermus thermophilus
0.15
-
 D-mannose mutant enzyme D256R, at pH 7.0 and 85°C Thermus thermophilus
0.17
-
 D-Lyxose mutant enzyme D256R, at pH 7.0 and 85°C Thermus thermophilus