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Literature summary for 1.1.1.39 extracted from

  • Zhang, Y.; Aono, T.; Poole, P.; Finan, T.M.
    NAD(P)+-malic enzyme mutants of Sinorhizobium sp. strain NGR234, but not Azorhizobium caulinodans ORS571, maintain symbiotic N2 fixation capabilities (2012), Appl. Environ. Microbiol., 78, 2803-2812.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
fumarate
-
Azorhizobium caulinodans
fumarate
-
Sinorhizobium sp.
succinate
-
Azorhizobium caulinodans
succinate
-
Sinorhizobium sp.

Cloned(Commentary)

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

Protein Variants Comment Organism
additional information azc3656 mutants show about 4fold reduced NAD+-malic enzyme activity Sinorhizobium sp.

Inhibitors

Inhibitors Comment Organism Structure
acetyl-CoA
-
Azorhizobium caulinodans
acetyl-CoA
-
Sinorhizobium sp.

KM Value [mM]

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Azorhizobium caulinodans
-
gene azc3656
-
Sinorhizobium sp. B6E9W4 gene azc3656 or dme
-
Sinorhizobium sp. NGR234 B6E9W4 gene azc3656 or dme
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme AZC3656 from Escherichia coli by nickel affinity chromatography Sinorhizobium sp.

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Sinorhizobium sp.

pH Optimum

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

Cofactor Comment Organism Structure
NAD+
-
Sinorhizobium sp.
NAD+ preferred cofactor Azorhizobium caulinodans
NADP+
-
Azorhizobium caulinodans
NADP+
-
Sinorhizobium sp.

General Information

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