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Literature summary extracted from

  • Gao, X.; Huang, F.; Feng, J.; Chen, X.; Zhang, H.; Wang, Z.; Wu, Q.; Zhu, D.
    Engineering the meso-diaminopimelate dehydrogenase from Symbiobacterium thermophilum by site saturation mutagenesis for D-phenylalanine synthesis (2013), Appl. Environ. Microbiol., 79, 5078-5081.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.1.16 recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Symbiobacterium thermophilum

Protein Variants

EC Number Protein Variants Comment Organism
1.4.1.16 H227C site-directed saturation mutagenesis, the mutant shows 15.1fold increased activity with phenylpyruvate compared to the wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 H227V site-directed saturation mutagenesis, the mutant shows 35.1fold increased activity with phenylpyruvate compared to the wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 additional information in order to enlarge the substrate binding pocket of the meso-diaminopimelate dehydrogenase from Symbiobacterium thermophilum to accommodate larger 2-keto acids, e.g. phenylalanine, four amino acid residues, Phe146, Thr171, Arg181, and His227, are targeted for site saturation mutagenesis Symbiobacterium thermophilum
1.4.1.16 R181F site-directed saturation mutagenesis, the mutant shows 6.4fold increased activity with phenylpyruvate compared to the wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 R181F/H227V site-directed saturation mutagenesis, the mutant shows 19.3fold increased activity with phenylpyruvate compared to the wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 T171P site-directed saturation mutagenesis, the mutant shows 2.2fold increased activity with phenylpyruvate compared to the wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 T171S site-directed saturation mutagenesis, the mutant shows 2.8fold increased activity with phenylpyruvate compared to the wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 T171S/H227V site-directed saturation mutagenesis, the mutant shows 10.6fold increased activity with phenylpyruvate compared to the wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 T171S/R181F site-directed saturation mutagenesis, the mutant shows 4.0fold increased activity with phenylpyruvate compared to the wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 T171S/R181F/H227V site-directed saturation mutagenesis, the mutant shows 14.6fold increased activity with phenylpyruvate compared to the wild-type enzyme Symbiobacterium thermophilum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.1.16 11
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant T171S/R181F/H227V Symbiobacterium thermophilum
1.4.1.16 11.1
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant R181F Symbiobacterium thermophilum
1.4.1.16 12.5
-
 phenylpyruvate pH 8.5, 30°C, recombinant wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 13.9
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant T171S/H227V Symbiobacterium thermophilum
1.4.1.16 13.9
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant T171S/R181F Symbiobacterium thermophilum
1.4.1.16 15.5
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant H227C Symbiobacterium thermophilum
1.4.1.16 15.8
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutantT171P Symbiobacterium thermophilum
1.4.1.16 19.6
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant T171S Symbiobacterium thermophilum
1.4.1.16 20.8
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant R181F/H227V Symbiobacterium thermophilum
1.4.1.16 24.3
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant H227V Symbiobacterium thermophilum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.4.1.16 meso-2,6-diaminoheptanedioate + H2O + NADP+ Symbiobacterium thermophilum
-
 L-2-amino-6-oxoheptanedioate + NH3 + NADPH + H+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.4.1.16 Symbiobacterium thermophilum Q67PI3
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.1.16 recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) Symbiobacterium thermophilum

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4.1.16 0.07
-
 recombinant wild-type enzyme, pH 8.5, 30°C Symbiobacterium thermophilum
1.4.1.16 0.15
-
 recombinant mutant T171P, pH 8.5, 30°C Symbiobacterium thermophilum
1.4.1.16 0.19
-
 recombinant mutant T171S, pH 8.5, 30°C Symbiobacterium thermophilum
1.4.1.16 0.28
-
 recombinant mutant T171S/R181F, pH 8.5, 30°C Symbiobacterium thermophilum
1.4.1.16 0.44
-
 recombinant mutant R181F, pH 8.5, 30°C Symbiobacterium thermophilum
1.4.1.16 0.74
-
 recombinant mutant T171S/H227V, pH 8.5, 30°C Symbiobacterium thermophilum
1.4.1.16 1.02
-
 recombinant mutant T171S/R181F/H227V, pH 8.5, 30°C Symbiobacterium thermophilum
1.4.1.16 1.03
-
 recombinant mutant H227C, pH 8.5, 30°C Symbiobacterium thermophilum
1.4.1.16 1.35
-
 recombinant mutant R181F/H227V, pH 8.5, 30°C Symbiobacterium thermophilum
1.4.1.16 2.39
-
 recombinant mutant H227V, pH 8.5, 30°C Symbiobacterium thermophilum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.1.16 meso-2,6-diaminoheptanedioate + H2O + NADP+
-
 Symbiobacterium thermophilum L-2-amino-6-oxoheptanedioate + NH3 + NADPH + H+
-
 r
1.4.1.16 phenylpyruvate + NH3 + NADPH + H+
-
 Symbiobacterium thermophilum phenylalanine + NADP+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.4.1.16 meso-DAPDH
-
 Symbiobacterium thermophilum
1.4.1.16 meso-diaminopimelate dehydrogenase
-
 Symbiobacterium thermophilum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.4.1.16 30
-
 assay at Symbiobacterium thermophilum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.1.16 0.11
-
 phenylpyruvate pH 8.5, 30°C, recombinant wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 0.53
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutantT171P Symbiobacterium thermophilum
1.4.1.16 0.7
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant T171S/R181F Symbiobacterium thermophilum
1.4.1.16 0.88
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant T171S Symbiobacterium thermophilum
1.4.1.16 1.07
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant R181F Symbiobacterium thermophilum
1.4.1.16 2.09
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant T171S/R181F/H227V Symbiobacterium thermophilum
1.4.1.16 2.37
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant T171S/H227V Symbiobacterium thermophilum
1.4.1.16 2.48
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant H227C Symbiobacterium thermophilum
1.4.1.16 3.43
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant R181F/H227V Symbiobacterium thermophilum
1.4.1.16 3.98
-
 phenylpyruvate pH 8.5, 30°C, recombinant mutant H227V Symbiobacterium thermophilum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.1.16 8.5
-
 assay at Symbiobacterium thermophilum

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.1.16 NADP+
-
 Symbiobacterium thermophilum
1.4.1.16 NADPH
-
 Symbiobacterium thermophilum

General Information

EC Number General Information Comment Organism
1.4.1.16 additional information molecular dynamic simulation analysis of wild-type and variant H227V enzyme with the imine intermediate as the substrate, overview Symbiobacterium thermophilum