Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Scheffersomyces stipitis |
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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Scheffersomyces stipitis | P22144 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Scheffersomyces stipitis |
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 | 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 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 | 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 |