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Literature summary extracted from

  • Gao, X.; Chen, X.; Liu, W.; Feng, J.; Wu, Q.; Hua, L.; Zhu, D.
    A novel meso-Diaminopimelate dehydrogenase from Symbiobacterium thermophilum: overexpression, characterization, and potential for D-amino acid synthesis (2012), Appl. Environ. Microbiol., 78, 8595-8600.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.4.1.16 biotechnology high thermostability and relaxed substrate profile of Symbiobacterium thermophilum meso-DAPDH warrant it as an excellent starting enzyme for creating effective D-amino acid dehydrogenases by protein engineering Symbiobacterium thermophilum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.1.16 DNA and amino acid sequence determmination and analysis, expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3), equally as a soluble protein and in inclusion bodies Symbiobacterium thermophilum

Protein Variants

EC Number Protein Variants Comment Organism
1.4.1.16 F146W site-directed mutagenesis, the mutant shows altered activity levels with meso-2,6-diaminoheptanedioate and pyruvate as substrates Symbiobacterium thermophilum
1.4.1.16 F146W/M152Q site-directed mutagenesis, the mutant shows altered activity levels with meso-2,6-diaminoheptanedioate and pyruvate as substrates Symbiobacterium thermophilum
1.4.1.16 M152Q site-directed mutagenesis, the mutant shows altered activity levels with meso-2,6-diaminoheptanedioate and pyruvate as substrates Symbiobacterium thermophilum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.1.16 1.9
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 2.5
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant mutant F1446W Symbiobacterium thermophilum
1.4.1.16 12.2
-
 pyruvate pH 8.5, 30°C, recombinant mutant F1446W Symbiobacterium thermophilum
1.4.1.16 13.5
-
 pyruvate pH 8.5, 30°C, recombinant wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 15.7
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant mutant F146W/M152Q Symbiobacterium thermophilum
1.4.1.16 16.7
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant mutant M152Q Symbiobacterium thermophilum
1.4.1.16 23.6
-
 pyruvate pH 8.5, 30°C, recombinant mutant F146W/M152Q Symbiobacterium thermophilum
1.4.1.16 24.8
-
 pyruvate pH 8.5, 30°C, recombinant mutant M152Q 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
1.4.1.16 additional information Symbiobacterium thermophilum the enzyme has a more relaxed substrate specificity and potential for D-amino acid synthesis ?
-
 ?

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 soluble enzyme 3.4fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography 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 additional information the enzyme has a more relaxed substrate specificity and potential for D-amino acid synthesis Symbiobacterium thermophilum ?
-
 ?
1.4.1.16 additional information meso-diaminopimelate dehydrogenase is an NADP-dependent enzyme which catalyzes the reversible oxidative deamination on the D-configuration of meso-2,6-diaminopimelate to produce L-2-amino-6-oxopimelate. The purified enzyme also shows activity toward D-alanine, D-valine, and D-lysine. This enzyme catalyzes the reductive amination of 2-keto acids such as pyruvic acid to generate D-amino acids in up to 99% conversion and 99% enantiomeric excess. Substrate specificities of recombinant wild-type and mutant emzymes in both amination and deamination, overview Symbiobacterium thermophilum ?
-
 ?
1.4.1.16 pyruvate + NH3 + NADPH + H+
-
 Symbiobacterium thermophilum D-alanine + 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

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.4.1.16 30 60 purified recombinant enzyme, 30 min, fully stable in deamination activity Symbiobacterium thermophilum
1.4.1.16 65
-
 purified recombinant enzyme, 30, loss of 50% amination activity Symbiobacterium thermophilum
1.4.1.16 70
-
 purified recombinant enzyme, 30, loss of 6% deamination activity Symbiobacterium thermophilum
1.4.1.16 75
-
 purified recombinant enzyme, 1 h, rapid inactivation Symbiobacterium thermophilum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.1.16 664
-
 pyruvate pH 8.5, 30°C, recombinant mutant F146W/M152Q Symbiobacterium thermophilum
1.4.1.16 1010
-
 pyruvate pH 8.5, 30°C, recombinant mutant M152Q Symbiobacterium thermophilum
1.4.1.16 6780
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant mutant F146W/M152Q Symbiobacterium thermophilum
1.4.1.16 7850
-
 pyruvate pH 8.5, 30°C, recombinant wild-type enzyme Symbiobacterium thermophilum
1.4.1.16 10100
-
 pyruvate pH 8.5, 30°C, recombinant mutant F1446W Symbiobacterium thermophilum
1.4.1.16 15100
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant mutant F1446W Symbiobacterium thermophilum
1.4.1.16 35700
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant mutant M152Q Symbiobacterium thermophilum
1.4.1.16 127000
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant wild-type enzyme Symbiobacterium thermophilum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.1.16 9
-
 amination Symbiobacterium thermophilum
1.4.1.16 11.5
-
 deamination 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 amino acid residues F146 and M152 are located in the substrate binding site, but are not critical in determining the enzyme's substrate specificity Symbiobacterium thermophilum