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Literature summary for 1.4.1.21 extracted from

  • Li, Y.; Kawakami, N.; Ogola, H.J.; Ashida, H.; Ishikawa, T.; Shibata, H.; Sawa, Y.
    A novel L-aspartate dehydrogenase from the mesophilic bacterium Pseudomonas aeruginosa PAO1: molecular characterization and application for L-aspartate production (2011), Appl. Microbiol. Biotechnol., 90, 1953-1962.
    View publication on PubMed
Search for more literature for 1.4.1.21

Cloned(Commentary)

Cloned (Comment) Organism
ORF PA3505, DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strain BL21 (DE3) Pseudomonas aeruginosa

Protein Variants

Protein Variants Comment Organism
additional information establishing of a L-Asp production system consisting of PaeAspDH, Bacillus subtilis malate dehydrogenase and Escherichia coli fumarase, achieving a high level of L-Asp production from fumarate in fed-batch process with a molar conversion yield of 89.4% in LB medium supplemented with fumarate, and 100 mM NH4Cl, overview, or in the same production system with glucose M9 minimal medium containing 50 mM glucose and 80 mM urea as carbon and nitrogen source, respectively Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Pseudomonas aeruginosa
0.045
-
 NADH pH 8.2, 37°C Pseudomonas aeruginosa
0.052
-
 NADPH pH 8.2, 37°C Pseudomonas aeruginosa
0.47
-
 NAD+ pH 9.8, 37°C Pseudomonas aeruginosa
0.47
-
 NADP+ pH 9.8, 37°C Pseudomonas aeruginosa
2.12
-
 oxaloacetate pH 8.2, 37°C, with NADH Pseudomonas aeruginosa
3.14
-
 oxaloacetate pH 8.2, 37°C, with NADPH Pseudomonas aeruginosa
4.74
-
 L-aspartate pH 9.8, 37°C, with NADP+ Pseudomonas aeruginosa
4.87
-
 L-aspartate pH 9.8, 37°C, with NAD+ Pseudomonas aeruginosa
10.1
-
 NH3 pH 8.2, 37°C, with NADH Pseudomonas aeruginosa
12.7
-
 NH3 pH 8.2, 37°C, with NADPH Pseudomonas aeruginosa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
27920
-
 2 * 27920, sequence calculation, 2 * 28000, SDS-PAGE Pseudomonas aeruginosa
28000
-
 2 * 27920, sequence calculation, 2 * 28000, SDS-PAGE Pseudomonas aeruginosa
60000
-
 gel filtration Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-aspartate + H2O + NAD(P)+ Pseudomonas aeruginosa
-
 oxaloacetate + NH3 + NAD(P)H + H+
-
 r

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa
-
 ORF PA3505
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli strain BL21 (DE3) to homogeneity Pseudomonas aeruginosa

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
127
-
 purified enzyme, substrate L-aspartate, pH 8.2, 37°C Pseudomonas aeruginosa
147
-
 purified enzyme, substrate oxaloacetate, pH 8.2, 37°C Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-aspartate + H2O + NAD(P)+
-
 Pseudomonas aeruginosa oxaloacetate + NH3 + NAD(P)H + H+
-
 r
L-aspartate + H2O + NAD+ the enzyme is capable of utilizing both NAD/H and NADP/H as coenzymes Pseudomonas aeruginosa oxaloacetate + NH3 + NADH + H+
-
 r
L-aspartate + H2O + NADP+ the enzyme is capable of utilizing both NAD/H and NADP/H as coenzymes Pseudomonas aeruginosa oxaloacetate + NH3 + NADPH + H+
-
 r
additional information the enzyme exhibits a very high specific activity for L-aspartate and oxaloacetate Pseudomonas aeruginosa ?
-
 ?
oxaloacetate + NH3 + NADH + H+ the enzyme is capable of utilizing both NAD/H and NADP/H as coenzymes Pseudomonas aeruginosa L-aspartate + H2O + NAD+
-
 r
oxaloacetate + NH3 + NADPH + H+ the enzyme is capable of utilizing both NAD/H and NADP/H as coenzymes Pseudomonas aeruginosa L-aspartate + H2O + NADP+
-
 r

Subunits

Subunits Comment Organism
dimer 2 * 27920, sequence calculation, 2 * 28000, SDS-PAGE Pseudomonas aeruginosa
More enzyme homology modeling, overview Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
L-aspDH
-
 Pseudomonas aeruginosa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Pseudomonas aeruginosa

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
48
-
 20 min, Tm of purified enzyme Pseudomonas aeruginosa
60
-
 20 min, Tm of purified enzyme in presence of 0.4 M NaCl or 30% glycerol Pseudomonas aeruginosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
42.7
-
 L-aspartate pH 9.8, 37°C, with NADP+ Pseudomonas aeruginosa
46.1
-
 NADP+ pH 9.8, 37°C Pseudomonas aeruginosa
59.7
-
 L-aspartate pH 9.8, 37°C, with NAD+ Pseudomonas aeruginosa
62
-
 NH3 pH 8.2, 37°C, with NADH Pseudomonas aeruginosa
62.9
-
 NAD+ pH 9.8, 37°C Pseudomonas aeruginosa
68.4
-
 oxaloacetate pH 8.2, 37°C, with NADH Pseudomonas aeruginosa
70.5
-
 NADH pH 8.2, 37°C Pseudomonas aeruginosa
71.6
-
 oxaloacetate pH 8.2, 37°C, with NADPH Pseudomonas aeruginosa
77.6
-
 NH3 pH 8.2, 37°C, with NADPH Pseudomonas aeruginosa
80.2
-
 NADPH pH 8.2, 37°C Pseudomonas aeruginosa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.2
-
 amination Pseudomonas aeruginosa
9.8
-
 deamination Pseudomonas aeruginosa

Cofactor

Cofactor Comment Organism Structure
additional information the enzyme is capable of utilizing both NAD/H and NADP/H as coenzymes Pseudomonas aeruginosa
NAD+
-
 Pseudomonas aeruginosa
NADH
-
 Pseudomonas aeruginosa
NADP+
-
 Pseudomonas aeruginosa
NADPH
-
 Pseudomonas aeruginosa

General Information

General Information Comment Organism
evolution L-aspartate dehydrogenase is a rare member of amino acid dehydrogenase superfamily Pseudomonas aeruginosa

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.11
-
 NH3 pH 8.2, 37°C, with NADPH Pseudomonas aeruginosa
6.14
-
 NH3 pH 8.2, 37°C, with NADH Pseudomonas aeruginosa
9.01
-
 L-aspartate pH 9.8, 37°C, with NADP+ Pseudomonas aeruginosa
12.3
-
 L-aspartate pH 9.8, 37°C, with NAD+ Pseudomonas aeruginosa
22.8
-
 oxaloacetate pH 8.2, 37°C, with NADPH Pseudomonas aeruginosa
32.3
-
 oxaloacetate pH 8.2, 37°C, with NADH Pseudomonas aeruginosa
98.1
-
 NADP+ pH 9.8, 37°C Pseudomonas aeruginosa
133.8
-
 NAD+ pH 9.8, 37°C Pseudomonas aeruginosa
1542
-
 NADPH pH 8.2, 37°C Pseudomonas aeruginosa
1567
-
 NADH pH 8.2, 37°C Pseudomonas aeruginosa