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Literature summary for 1.1.1.9 extracted from

  • Watanabe, S.; Kodaki, T.; Makino, K.
    Complete reversal of coenzyme specificity of xylitol dehydrogenase and increase of thermostability by the introduction of structural zinc (2005), J. Biol. Chem., 280, 10340-10349.
    View publication on PubMed
Search for more literature for 1.1.1.9

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Scheffersomyces stipitis

Protein Variants

Protein Variants Comment Organism
D207A kcat/Km for NAD+ is 3.6fold lower than wild-type value, kcat/Km for NADP+ is 4.3fold higher than wild-type value Scheffersomyces stipitis
D207A/F209S kcat/Km for NAD+ is 2.2fold lower than wild-type value, kcat/Km for NADP+ is 745fold higher than wild-type value Scheffersomyces stipitis
D207A/I208R kcat/Km for NAD+ is 2.5fold lower than wild-type value, kcat/Km for NADP+ is 229fold higher than wild-type value Scheffersomyces stipitis
D207A/I208R/F209S kcat/Km for NAD+ is 15.2fold lower than wild-type value, kcat/Km for NADP+ is 4292fold higher than wild-type value, increased thermostability Scheffersomyces stipitis
D207A/I208R/F209S/N211R kcat/Km for NAD+ is 32.9fold lower than wild-type value, kcat/Km for NADP+ is 4292fold higher than wild-type value, increased thermostability Scheffersomyces stipitis
D207A/I208R/F209T kcat/Km for NAD+ is 2.4fold lower than wild-type value, kcat/Km for NADP+ is 4754fold higher than wild-type value Scheffersomyces stipitis
D207A/I208R/F209Y kcat/Km for NAD+ is 6.9fold lowerthan wild-type value, kcat/Km for NADP+ is 788fold higher than wild-type value Scheffersomyces stipitis
F209S kcat/Km for NAD+ is 1.9fold lower than wild-type value, kcat/Km for NADP+ is 31.4fold higher than wild-type value Scheffersomyces stipitis
I208R kcat/Km for NAD+ is nearly identical to wild-type value, kcat/Km for NADP+ is 44fold higher than wild-type value Scheffersomyces stipitis
I208R/F209S kcat/Km for NAD+ is 30.7fold lower than wild-type value, kcat/Km for NADP+ is 1.5fold higher than wild-type value Scheffersomyces stipitis
N211R kcat/Km for NAD+ is 1.1fold lower than wild-type value, kcat/Km for NADP+ is 7.6fold higher than wild-type value Scheffersomyces stipitis
S96C/S99C/Y102C/D207A/I208R/F209S kcat/Km for NAD+ is 36.7fold lower than wild-type value, kcat/Km for NADP+ 16462is fold higher than wild-type value Scheffersomyces stipitis
S96C/S99C/Y102C/D207A/I208R/F209S/N211R kcat/Km for NAD+ is 26.5fold lower than wild-type value, kcat/Km for NADP+ is 16154fold higher than wild-type value Scheffersomyces stipitis
S96C/S99CY102C kcat/Km for NAD+ is 1.1fold lower than wild-type value, kcat/Km for NADP+ is 8.8fold higher than wild-type value Scheffersomyces stipitis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.152
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209Y Scheffersomyces stipitis
0.162
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/F209S Scheffersomyces stipitis
0.265
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209T Scheffersomyces stipitis
0.381
-
 NAD+ pH 9.0, 35°, wild-type enzyme Scheffersomyces stipitis
0.403
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A Scheffersomyces stipitis
0.498
-
 NAD+ pH 9.0, 35°C, mutant enzyme I208R Scheffersomyces stipitis
0.538
-
 NAD+ pH 9.0, 35°C, mutant enzyme N211R Scheffersomyces stipitis
0.568
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R Scheffersomyces stipitis
0.638
-
 xylitol pH 9.0, 35°C, cofactor: NADP+, mutant enzyme D207A/I208R/F209T Scheffersomyces stipitis
0.638
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209T Scheffersomyces stipitis
0.665
-
 NAD+ pH 9.0, 35°C, mutant enzyme I208R/F209S Scheffersomyces stipitis
0.731
-
 xylitol pH 9.0, 35°C, cofactor: NADP+, mutant enzyme D207A/I208R/F209Y Scheffersomyces stipitis
0.731
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209Y Scheffersomyces stipitis
0.739
-
 NAD+ pH 9.0, 35°C, mutant enzyme S96C/S99CY102C Scheffersomyces stipitis
0.848
-
 NAD+ pH 9.0, 35°C, mutant enzyme F209S Scheffersomyces stipitis
0.897
-
 xylitol pH 9.0, 35°C, cofactor: NADP+, mutant enzyme D207A/I208R/F209S Scheffersomyces stipitis
0.897
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209S Scheffersomyces stipitis
1.04
-
 xylitol pH 9.0, 35°C, cofactor: NADP+, mutant enzyme S96C/S99C/Y102C/D207A/I208R/F209S/N211R Scheffersomyces stipitis
1.04
-
 NADP+ pH 9.0, 35°C, mutant enzym S96C/S99C/Y102C/D207A/I208R/F209S/N211R Scheffersomyces stipitis
1.18
-
 xylitol pH 9.0, 35°C, cofactor: NADP+, mutant enzyme S96C/S99C/Y102C/D207A/I208R/F209S Scheffersomyces stipitis
1.18
-
 NADP+ pH 9.0, 35°C, mutant enzyme S96C/S99C/Y102C/D207A/I208R/F209S Scheffersomyces stipitis
1.3
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209S Scheffersomyces stipitis
1.38
-
 xylitol pH 9.0, 35°C, cofactor: NADP+, mutant enzyme D207A/I208R/F209S/N211R Scheffersomyces stipitis
1.38
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209S/N211R Scheffersomyces stipitis
4 5.4 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme I208R/F209S Scheffersomyces stipitis
7.6
-
 NAD+ pH 9.0, 35°C, mutant enzym S96C/S99C/Y102C/D207A/I208R/F209S/N211R Scheffersomyces stipitis
9.19
-
 NADP+ pH 9.0, 35°C, mutant enzyme I208R/F209S Scheffersomyces stipitis
9.56
-
 xylitol pH 9.0, 35°C, cofactor: NADP+, mutant enzyme S96C/S99CY102C Scheffersomyces stipitis
9.56
-
 NADP+ pH 9.0, 35°C, mutant enzyme S96C/S99CY102C Scheffersomyces stipitis
9.96
-
 xylitol pH 9.0, 35°C, cofactor: NADP+, mutant enzyme D207A/F209S Scheffersomyces stipitis
9.96
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/F209S Scheffersomyces stipitis
11.3
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R Scheffersomyces stipitis
17.3
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209S/N211R Scheffersomyces stipitis
21.1
-
 NADP+ pH 9.0, 35°C, mutant enzyme I208R Scheffersomyces stipitis
21.7
-
 xylitol pH 9.0, 35°, cofactor: NAD+, wild-type enzyme Scheffersomyces stipitis
22.2
-
 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme D207A Scheffersomyces stipitis
23.5
-
 NAD+ pH 9.0, 35°C, mutant enzyme S96C/S99C/Y102C/D207A/I208R/F209S Scheffersomyces stipitis
24.2
-
 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme D207A/I208R Scheffersomyces stipitis
27.4
-
 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme N211R Scheffersomyces stipitis
28.9
-
 NADP+ pH 9.0, 35°C, mutant enzyme F209S Scheffersomyces stipitis
29.5
-
 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme I208R Scheffersomyces stipitis
30.3
-
 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme S96C/S99CY102C Scheffersomyces stipitis
34.1
-
 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme F209S Scheffersomyces stipitis
45.2
-
 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme D207A/F209S Scheffersomyces stipitis
50.1
-
 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme D207A/I208R/F209Y Scheffersomyces stipitis
55.7
-
 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme D207A/I208R/F209S Scheffersomyces stipitis
56.5
-
 NADP+ pH 9.0, 35°C, mutant enzyme N211R Scheffersomyces stipitis
97.8
-
 xylitol pH 9.0, 35°C, cofactor: NAD+, mutant enzyme D207A/I208R/F209T Scheffersomyces stipitis
120
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A Scheffersomyces stipitis
170
-
 NADP+ pH 9.0, 35°, wild-type enzyme Scheffersomyces stipitis

Organism

Organism UniProt Comment Textmining
Scheffersomyces stipitis P22144
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Scheffersomyces stipitis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 specific activity of wild-type and mutant enzymes Scheffersomyces stipitis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
xylitol + NAD+
-
 Scheffersomyces stipitis D-xylulose + NADH + H+
-
 ?
xylitol + NADP+
-
 Scheffersomyces stipitis D-xylulose + NADPH + H+
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.017
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209T Scheffersomyces stipitis
0.052 2.1 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R Scheffersomyces stipitis
0.145
-
 NADP+ pH 9.0, 35°C, mutant enzyme I208R/F209S Scheffersomyces stipitis
0.5
-
 NAD+ pH 9.0, 35°C, mutant enzym S96C/S99C/Y102C/D207A/I208R/F209S/N211R Scheffersomyces stipitis
0.91
-
 NADP+ pH 9.0, 35°C, mutant enzyme S96C/S99CY102C Scheffersomyces stipitis
1
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209Y Scheffersomyces stipitis
1
-
 NAD+ pH 9.0, 35°C, mutant enzyme I208R/F209S Scheffersomyces stipitis
1.83
-
 NADP+ pH 9.0, 35°, wild-type enzyme Scheffersomyces stipitis
2 8 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R Scheffersomyces stipitis
3.33
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/F209S Scheffersomyces stipitis
4
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209S Scheffersomyces stipitis
4.67
-
 NADP+ pH 9.0, 35°C, mutant enzyme N211R Scheffersomyces stipitis
5.17
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A Scheffersomyces stipitis
5.17
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209T Scheffersomyces stipitis
5.7
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A Scheffersomyces stipitis
6.17
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209Y Scheffersomyces stipitis
8
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/F209S Scheffersomyces stipitis
9.83
-
 NADP+ pH 9.0, 35°C, mutant enzyme F209S Scheffersomyces stipitis
10
-
 NADP+ pH 9.0, 35°C, mutant enzyme I208R Scheffersomyces stipitis
10.33
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R Scheffersomyces stipitis
13.17
-
 NAD+ pH 9.0, 35°C, mutant enzym S96C/S99C/Y102C/D207A/I208R/F209S/N211R Scheffersomyces stipitis
16.9
-
 NAD+ pH 9.0, 35°C, mutant enzyme N211R Scheffersomyces stipitis
17.5
-
 NAD+ pH 9.0, 35°, wild-type enzyme Scheffersomyces stipitis
18
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209S/N211R Scheffersomyces stipitis
20.33
-
 NAD+ pH 9.0, 35°C, mutant enzyme F209S Scheffersomyces stipitis
22.83
-
 NAD+ pH 9.0, 35°C, mutant enzyme N211R Scheffersomyces stipitis
23.8
-
 NAD+ pH 9.0, 35°C, mutant enzyme I208R Scheffersomyces stipitis
23.83
-
 NAD+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209S/N211R Scheffersomyces stipitis
29.5
-
 NAD+ pH 9.0, 35°C, mutant enzyme S96C/S99C/Y102C/D207A/I208R/F209S Scheffersomyces stipitis
30.33
-
 NAD+ pH 9.0, 35°C, mutant enzyme S96C/S99CY102C Scheffersomyces stipitis
32.83
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209T Scheffersomyces stipitis
41.7
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209S Scheffersomyces stipitis
64
-
 NADP+ pH 9.0, 35°C, mutant enzyme D207A/I208R/F209S/N211R Scheffersomyces stipitis
183.3
-
 NADP+ pH 9.0, 35°C, mutant enzym S96C/S99C/Y102C/D207A/I208R/F209S/N211R Scheffersomyces stipitis
210
-
 NADP+ pH 9.0, 35°C, mutant enzyme S96C/S99C/Y102C/D207A/I208R/F209S Scheffersomyces stipitis

Cofactor

Cofactor Comment Organism Structure
NAD+ kcat/Km for wild-type enzyme is 2760/min*mM Scheffersomyces stipitis
NADP+ kcat/Km for wild-type enzyme is 0.65/min*mM Scheffersomyces stipitis