Application | Comment | Organism |
---|---|---|
synthesis | overexpressing NAD kinase is a useful metabolic engineering strategy to improve NADPH supply and isoleucine biosynthesis | Corynebacterium glutamicum |
synthesis | overexpressing NAD kinase is a useful metabolic engineering strategy to improve NADPH supply and isoleucine biosynthesis | Corynebacterium glutamicum subsp. lactofermentum |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Corynebacterium glutamicum |
expressed in Escherichia coli BL21(DE3) cells | Corynebacterium glutamicum subsp. lactofermentum |
Protein Variants | Comment | Organism |
---|---|---|
P117S | the mutation results in increased ATP-NAD+ kinase activity and ATP-NADH kinase activity | Corynebacterium glutamicum |
P117S | the mutation results in increased ATP-NAD+ kinase activity and ATP-NADH kinase activity | Corynebacterium glutamicum subsp. lactofermentum |
P57S | the mutation results in increased ATP-NAD+ kinase activity and ATP-NADH kinase activity | Corynebacterium glutamicum |
P57S | the mutation results in increased ATP-NAD+ kinase activity and ATP-NADH kinase activity | Corynebacterium glutamicum subsp. lactofermentum |
P57S/P117S | the double mutation only exhibits a little higher activity than P117S-single point mutation. The catalytic efficiency of the mutant improves greatly, which is 6.8 (for NAD+) and 3.2fold (for ATP) higher than that of the wild type enzyme | Corynebacterium glutamicum |
P57S/P117S | the double mutation only exhibits a little higher activity than P117S-single point mutation. The catalytic efficiency of the mutant improves greatly, which is 6.8 (for NAD+) and 3.2fold (for ATP) higher than that of the wild type enzyme | Corynebacterium glutamicum subsp. lactofermentum |
S57P | the variation is associated with the decreased enzyme activity | Corynebacterium glutamicum |
S57P | the variation is associated with the decreased enzyme activity | Corynebacterium glutamicum subsp. lactofermentum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Corynebacterium glutamicum | |
NADH | - |
Corynebacterium glutamicum subsp. lactofermentum | |
NADP+ | - |
Corynebacterium glutamicum | |
NADP+ | - |
Corynebacterium glutamicum subsp. lactofermentum | |
NADPH | - |
Corynebacterium glutamicum | |
NADPH | - |
Corynebacterium glutamicum subsp. lactofermentum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.07 | - |
ATP | mutant enzyme P57S/P117S, at pH 6.0 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
1.4 | - |
NAD+ | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum | |
1.41 | - |
NAD+ | mutant enzyme P57S/P117S, at pH 6.0 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
1.95 | - |
ATP | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
2.11 | - |
NAD+ | mutant enzyme P57S/P117S, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
2.15 | - |
ATP | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum | |
3.73 | - |
ATP | mutant enzyme P57S/P117S, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
4.02 | - |
NAD+ | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
36000 | - |
4 * 36000, SDS-PAGE | Corynebacterium glutamicum |
36000 | - |
4 * 36000, SDS-PAGE | Corynebacterium glutamicum subsp. lactofermentum |
140000 | - |
gel filtration | Corynebacterium glutamicum |
140000 | - |
gel filtration | Corynebacterium glutamicum subsp. lactofermentum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium glutamicum | - |
subsp. lactofermentum | - |
Corynebacterium glutamicum ATCC 13869 | - |
subsp. lactofermentum | - |
Corynebacterium glutamicum subsp. lactofermentum | - |
- |
- |
Corynebacterium glutamicum subsp. lactofermentum JHI3-156 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography, gel filtration | Corynebacterium glutamicum |
Ni-NTA column chromatography, gel filtration | Corynebacterium glutamicum subsp. lactofermentum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + NAD+ | ATP is the preferred phosphoryl donor and NAD+ is the preferred acceptor | Corynebacterium glutamicum | ADP + NADP+ | - |
? | |
ATP + NAD+ | ATP is the preferred phosphoryl donor and NAD+ is the preferred acceptor | Corynebacterium glutamicum subsp. lactofermentum | ADP + NADP+ | - |
? | |
ATP + NAD+ | ATP is the preferred phosphoryl donor and NAD+ is the preferred acceptor | Corynebacterium glutamicum ATCC 13869 | ADP + NADP+ | - |
? | |
ATP + NAD+ | ATP is the preferred phosphoryl donor and NAD+ is the preferred acceptor | Corynebacterium glutamicum subsp. lactofermentum JHI3-156 | ADP + NADP+ | - |
? | |
hexaphosphate + NAD+ | - |
Corynebacterium glutamicum subsp. lactofermentum | pentaphosphate + NADP+ | - |
? | |
hexaphosphate + NAD+ | - |
Corynebacterium glutamicum subsp. lactofermentum JHI3-156 | pentaphosphate + NADP+ | - |
? | |
hexapolyphosphate + NAD+ | - |
Corynebacterium glutamicum | pentapolyphosphate + NADP+ | - |
? | |
hexapolyphosphate + NAD+ | - |
Corynebacterium glutamicum ATCC 13869 | pentapolyphosphate + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | 4 * 36000, SDS-PAGE | Corynebacterium glutamicum |
homotetramer | 4 * 36000, SDS-PAGE | Corynebacterium glutamicum subsp. lactofermentum |
Synonyms | Comment | Organism |
---|---|---|
NAD kinase | - |
Corynebacterium glutamicum |
NAD kinase | - |
Corynebacterium glutamicum subsp. lactofermentum |
poly(P)/ATPdependent NAD kinase | - |
Corynebacterium glutamicum |
poly(P)/ATPdependent NAD kinase | - |
Corynebacterium glutamicum subsp. lactofermentum |
Ppnk | - |
Corynebacterium glutamicum |
Ppnk | - |
Corynebacterium glutamicum subsp. lactofermentum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
preincubating the enzyme for 2 h at 70°C results in a loss of 50% of its activity | Corynebacterium glutamicum |
70 | - |
preheating the enzyme for 2 h at temperatures up to 70°C does not alter the ATP-NAD+ kinase activity significantly (less than 10% of the activity lost) | Corynebacterium glutamicum subsp. lactofermentum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.5 | - |
NAD+ | mutant enzyme P57S/P117S, at pH 6.0 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
0.52 | - |
ATP | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum | |
0.54 | - |
NAD+ | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum | |
0.6 | - |
ATP | mutant enzyme P57S/P117S, at pH 6.0 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
2.54 | - |
ATP | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
3.8 | - |
NAD+ | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
15.42 | - |
NAD+ | mutant enzyme P57S/P117S, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
20.51 | - |
ATP | mutant enzyme P57S/P117S, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Corynebacterium glutamicum subsp. lactofermentum |
8 | - |
- |
Corynebacterium glutamicum |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | - |
the enzyme remains stable at pH 7.0 | Corynebacterium glutamicum subsp. lactofermentum |
7 | 8 | the enzyme is precipitated and denatured at pH 7.0. Even at pH 8.0, the enzyme is depolymerized and denatured after long-time storing | Corynebacterium glutamicum |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.17 | - |
NADPH | at pH 8.0 and 30°C | Corynebacterium glutamicum | |
0.21 | - |
NADP+ | at pH 8.0 and 30°C | Corynebacterium glutamicum | |
0.22 | - |
NADPH | at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
0.24 | - |
NADH | at pH 8.0 and 30°C | Corynebacterium glutamicum | |
0.43 | - |
NADP+ | at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
0.62 | - |
NADH | at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum |
General Information | Comment | Organism |
---|---|---|
metabolism | NAD kinase is the key enzyme for the de novo biosynthesis of NADP+ and NADPH | Corynebacterium glutamicum |
metabolism | NAD kinase is the key enzyme for the de novo biosynthesis of NADP+ and NADPH | Corynebacterium glutamicum subsp. lactofermentum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.24 | - |
ATP | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum | |
0.36 | - |
NAD+ | mutant enzyme P57S/P117S, at pH 6.0 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
0.39 | - |
NAD+ | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum | |
0.56 | - |
ATP | mutant enzyme P57S/P117S, at pH 6.0 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
0.94 | - |
NAD+ | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
1.3 | - |
ATP | wild type enzyme, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
5.5 | - |
ATP | mutant enzyme P57S/P117S, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum | |
7.32 | - |
NAD+ | mutant enzyme P57S/P117S, at pH 7.5 and 30°C | Corynebacterium glutamicum subsp. lactofermentum |