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Literature summary for 1.2.1.11 extracted from

  • Xu, X.; Chen, J.; Wang, Q.; Duan, C.; Li, Y.; Wang, R.; Yang, S.
    Mutagenesis of key residues in the binding center of L-aspartate-beta-semialdehyde dehydrogenase from Escherichia coli enhances utilization of the cofactor NAD(H) (2016), ChemBioChem, 17, 56-64 .
    View publication on PubMed
Search for more literature for 1.2.1.11

Application

Application Comment Organism
biotechnology the modofied enzyme with altered substrate specificity using NAD(H) is preferred in biotechnological production of amino acids due to lower costs and higher stability Escherichia coli
synthesis the modofied enzyme with altered substrate specificity using NAD(H) is preferred in biotechnological production of amino acids due to lower costs and higher stability Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
gene asd, construction of a genetic ecASADH library by saturation mutagenesis, recombinant expression of His-tagged wild-type and selected mutants in Escherichia coli strain BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
A163S site-directed mutagenesis, the mutant shows almost unaltered cofactor specificity compared to the wild-type enzyme Escherichia coli
H171 site-directed mutagenesis, the mutant shows almost unaltered cofactor specificity compared to the wild-type enzyme Escherichia coli
L351V site-directed mutagenesis, the mutant shows unaltered cofactor specificity compared to the wild-type enzyme Escherichia coli
Q350N site-directed mutagenesis, the mutant shows 44fold increased activity with NAD+ compared to the wild-type enzyme and can also also utilize NADH efficiently. Unlike the wild-type enzyme, mutants Q350N and Q350N/H171A are able to synthesize L-homoserine from aspartate efficiently with NADH as a cofactor Escherichia coli
Q350N/H171A Q350N site-directed mutagenesis, the mutant shows 66fold increased activity with NAD+ compared to the wild-type enzyme and can also utilize NADH efficiently. Unlike the wild-type enzyme, mutants Q350N and Q350N/H171A are able to synthesize L-homoserine from aspartate efficiently with NADH as a cofactor Escherichia coli
S138Q site-directed mutagenesis, the mutant shows unaltered cofactor specificity compared to the wild-type enzyme Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0026
-
 NADP+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
0.0057
-
 NADP+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli
0.2
-
 NADP+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli
2.2
-
 NAD+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
2.5
-
 NAD+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli
11.4
-
 NAD+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-aspartate 4-semialdehyde + phosphate + NADP+ Escherichia coli
-
 L-4-aspartyl phosphate + NADPH + H+
-
 r

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A9Q9 MG1655
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutants from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-aspartate 4-semialdehyde + phosphate + NAD+
-
 Escherichia coli L-4-aspartyl phosphate + NADH + H+
-
 r
L-aspartate 4-semialdehyde + phosphate + NADP+
-
 Escherichia coli L-4-aspartyl phosphate + NADPH + H+
-
 r

Synonyms

Synonyms Comment Organism
ASADH
-
 Escherichia coli
L-aspartate-beta-semialdehyde dehydrogenase
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
9.5
-
 NAD+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli
13.4
-
 NADP+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
53.4
-
 NADP+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli
86.2
-
 NAD+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli
115.2
-
 NAD+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
258
-
 NADP+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
additional information cofactor modes of wild-type and mutant enzymes determined by molecular modeling Escherichia coli
NAD+ very low activity with the wild-type enzyme, but 44fold and 66fold higher activity with enzyme mutants Q350N and Q350N/H171A, respectively, compared to the wild-type Escherichia coli
NADH
-
 Escherichia coli
NADP+ highly preferred by the wild-type enzyme Escherichia coli
NADPH
-
 Escherichia coli

General Information

General Information Comment Organism
metabolism the enzyme has a rate-limiting key function in the biosynthesis of amino acids L-threonine, L-lysine, and L-isoleucine from L-aspartate via L-homoserine Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.8
-
 NAD+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli
35.1
-
 NAD+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli
53.3
-
 NAD+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
1124.9
-
 NADP+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli
5135.9
-
 NADP+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
9612.8
-
 NADP+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli