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Literature summary 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

Application

EC Number Application Comment Organism
1.2.1.11 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
1.2.1.11 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)

EC Number Cloned (Comment) Organism
1.2.1.11 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

EC Number Protein Variants Comment Organism
1.2.1.11 A163S site-directed mutagenesis, the mutant shows almost unaltered cofactor specificity compared to the wild-type enzyme Escherichia coli
1.2.1.11 H171 site-directed mutagenesis, the mutant shows almost unaltered cofactor specificity compared to the wild-type enzyme Escherichia coli
1.2.1.11 L351V site-directed mutagenesis, the mutant shows unaltered cofactor specificity compared to the wild-type enzyme Escherichia coli
1.2.1.11 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
1.2.1.11 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
1.2.1.11 S138Q site-directed mutagenesis, the mutant shows unaltered cofactor specificity compared to the wild-type enzyme Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.11 0.0026
-
 NADP+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
1.2.1.11 0.0057
-
 NADP+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli
1.2.1.11 0.2
-
 NADP+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli
1.2.1.11 2.2
-
 NAD+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
1.2.1.11 2.5
-
 NAD+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli
1.2.1.11 11.4
-
 NAD+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.11 Escherichia coli P0A9Q9 MG1655
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.2.1.11 30
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.2.1.11 9.5
-
 NAD+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli
1.2.1.11 13.4
-
 NADP+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
1.2.1.11 53.4
-
 NADP+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli
1.2.1.11 86.2
-
 NAD+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli
1.2.1.11 115.2
-
 NAD+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
1.2.1.11 258
-
 NADP+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.2.1.11 9
-
 assay at Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.11 additional information cofactor modes of wild-type and mutant enzymes determined by molecular modeling Escherichia coli
1.2.1.11 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
1.2.1.11 NADH
-
 Escherichia coli
1.2.1.11 NADP+ highly preferred by the wild-type enzyme Escherichia coli
1.2.1.11 NADPH
-
 Escherichia coli

General Information

EC Number General Information Comment Organism
1.2.1.11 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.2.1.11 0.8
-
 NAD+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli
1.2.1.11 35.1
-
 NAD+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli
1.2.1.11 53.3
-
 NAD+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
1.2.1.11 1124.9
-
 NADP+ recombinant wild-type enzyme, pH 9.0, 30°C Escherichia coli
1.2.1.11 5135.9
-
 NADP+ recombinant mutant Q350N/H171A, pH 9.0, 30°C Escherichia coli
1.2.1.11 9612.8
-
 NADP+ recombinant mutant Q350N, pH 9.0, 30°C Escherichia coli