Activating Compound | Comment | Organism | Structure |
---|---|---|---|
fumarate | - |
Azorhizobium caulinodans | |
fumarate | - |
Sinorhizobium sp. | |
succinate | - |
Azorhizobium caulinodans | |
succinate | - |
Sinorhizobium sp. |
Cloned (Comment) | Organism |
---|---|
gene dme or azc3656, DNA and amino acid sequence determination and analysis, expression as His-tagged enzyme in Escherichia coli | Sinorhizobium sp. |
Protein Variants | Comment | Organism |
---|---|---|
additional information | azc3656 mutants show about 4fold reduced NAD+-malic enzyme activity | Sinorhizobium sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
acetyl-CoA | - |
Azorhizobium caulinodans | |
acetyl-CoA | - |
Sinorhizobium sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.101 | - |
NAD+ | pH 7.8, 30°C, AZC3656 | Sinorhizobium sp. | |
0.59 | - |
(S)-malate | pH 7.8, 30°C, AZC3656, in presence of 1 mM fumarate | Sinorhizobium sp. | |
0.64 | - |
(S)-malate | pH 7.8, 30°C, AZC3656, in presence of 10 mM succinate | Sinorhizobium sp. | |
2.1 | - |
NADP+ | pH 7.8, 30°C, AZC3656 | Sinorhizobium sp. | |
2.7 | - |
(S)-malate | pH 7.8, 30°C, AZC3656 | Sinorhizobium sp. | |
27.6 | - |
(S)-malate | pH 7.8, 30°C, AZC3656, in presence of 0.05 mM acetyl-CoA | Sinorhizobium sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD(P)+ | Azorhizobium caulinodans | - |
pyruvate + CO2 + NAD(P)H | - |
? | |
(S)-malate + NAD+ | Sinorhizobium sp. | - |
pyruvate + CO2 + NADH | - |
? | |
(S)-malate + NAD+ | Sinorhizobium sp. NGR234 | - |
pyruvate + CO2 + NADH | - |
? | |
(S)-malate + NADP+ | Sinorhizobium sp. | - |
pyruvate + CO2 + NADPH | - |
? | |
(S)-malate + NADP+ | Sinorhizobium sp. NGR234 | - |
pyruvate + CO2 + NADPH | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Azorhizobium caulinodans | - |
gene azc3656 | - |
Sinorhizobium sp. | B6E9W4 | gene azc3656 or dme | - |
Sinorhizobium sp. NGR234 | B6E9W4 | gene azc3656 or dme | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme AZC3656 from Escherichia coli by nickel affinity chromatography | Sinorhizobium sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.201 | - |
NAD+-dependent ME activity in the wild-type strain, pH 7.8, 30°C, AZC3656 | Sinorhizobium sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD(P)+ | - |
Azorhizobium caulinodans | pyruvate + CO2 + NAD(P)H | - |
? | |
(S)-malate + NAD+ | - |
Sinorhizobium sp. | pyruvate + CO2 + NADH | - |
? | |
(S)-malate + NAD+ | - |
Sinorhizobium sp. NGR234 | pyruvate + CO2 + NADH | - |
? | |
(S)-malate + NADP+ | - |
Sinorhizobium sp. | pyruvate + CO2 + NADPH | - |
? | |
(S)-malate + NADP+ | - |
Sinorhizobium sp. NGR234 | pyruvate + CO2 + NADPH | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AZC3656 protein | - |
Azorhizobium caulinodans |
AZC3656 protein | - |
Sinorhizobium sp. |
DME | - |
Azorhizobium caulinodans |
DME | - |
Sinorhizobium sp. |
NAD(P)+-malic enzyme | - |
Azorhizobium caulinodans |
NAD(P)+-malic enzyme | - |
Sinorhizobium sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Sinorhizobium sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
AZC3656 shows high NAD+-ME activity at pH 7.8, with Michaelis-Menten-like kinetics, at various concentrations of malate, The enzyme exhibits only a very limited positive cooperativity with respect to malate | Sinorhizobium sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Sinorhizobium sp. | |
NAD+ | preferred cofactor | Azorhizobium caulinodans | |
NADP+ | - |
Azorhizobium caulinodans | |
NADP+ | - |
Sinorhizobium sp. |
General Information | Comment | Organism |
---|---|---|
malfunction | dme mutants of the broad-host-range Sinorhizobium sp. strain NGR234 form nodules whose level of N2 fixation vary from 27 to 83% (plant dry weight) of the wild-type level, depending on the host plant inoculated. The single dme mutant fixes N2 at reduced rate. A pckA dme double mutant has no N2-fixing activity, PCK is phosphoenolpyruvate carboxykinase. Symbiotic phenotypes of NGR234 and NGR234 dme mutants on different host plants, overview | Sinorhizobium sp. |
physiological function | AZC3656 protein is a NAD+-malic enzyme, i.e. DME, while AZC0119 protein is not a malic enzyme. DME is considered an important enzyme for regulating C4-dicarboxylic acid metabolism in N2-fixing bacteroids because its activity is strongly inhibited by acetyl-CoA and stimulated by fumarate and succinate. The NAD+-malic enzyme is required for N2 fixation, and this activity is thought to be required for the anaplerotic synthesis of pyruvate. But NGR234 bacteroids appear to synthesize pyruvate from TCA cycle intermediates via DME or PCK, phosphoenolpyruvate carboxykinase, pathways, overview | Sinorhizobium sp. |
physiological function | DME is considered an important enzyme for regulating C4-dicarboxylic acid metabolism in N2-fixing bacteroids because its activity is strongly inhibited by acetyl-CoA and stimulated by fumarate and succinate. The NAD+-malic enzyme is required for N2 fixation, and this activity is thought to be required for the anaplerotic synthesis of pyruvate | Azorhizobium caulinodans |