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Literature summary for 1.4.1.16 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
Search for more literature for 1.4.1.16

Application

Application Comment Organism
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)

Cloned (Comment) Organism
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

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.9
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant wild-type enzyme Symbiobacterium thermophilum
2.5
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant mutant F1446W Symbiobacterium thermophilum
12.2
-
 pyruvate pH 8.5, 30°C, recombinant mutant F1446W Symbiobacterium thermophilum
13.5
-
 pyruvate pH 8.5, 30°C, recombinant wild-type enzyme Symbiobacterium thermophilum
15.7
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant mutant F146W/M152Q Symbiobacterium thermophilum
16.7
-
 meso-2,6-diaminoheptanedioate pH 8.5, 30°C, recombinant mutant M152Q Symbiobacterium thermophilum
23.6
-
 pyruvate pH 8.5, 30°C, recombinant mutant F146W/M152Q Symbiobacterium thermophilum
24.8
-
 pyruvate pH 8.5, 30°C, recombinant mutant M152Q Symbiobacterium thermophilum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
meso-2,6-diaminoheptanedioate + H2O + NADP+ Symbiobacterium thermophilum
-
 L-2-amino-6-oxoheptanedioate + NH3 + NADPH + H+
-
 r
additional information Symbiobacterium thermophilum the enzyme has a more relaxed substrate specificity and potential for D-amino acid synthesis ?
-
 ?

Organism

Organism UniProt Comment Textmining
Symbiobacterium thermophilum Q67PI3
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant soluble enzyme 3.4fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Symbiobacterium thermophilum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
meso-2,6-diaminoheptanedioate + H2O + NADP+
-
 Symbiobacterium thermophilum L-2-amino-6-oxoheptanedioate + NH3 + NADPH + H+
-
 r
additional information the enzyme has a more relaxed substrate specificity and potential for D-amino acid synthesis Symbiobacterium thermophilum ?
-
 ?
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 ?
-
 ?
pyruvate + NH3 + NADPH + H+
-
 Symbiobacterium thermophilum D-alanine + NADP+
-
 ?

Synonyms

Synonyms Comment Organism
meso-DAPDH
-
 Symbiobacterium thermophilum
meso-diaminopimelate dehydrogenase
-
 Symbiobacterium thermophilum

Temperature Optimum [°C]

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

Temperature Stability [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
 amination Symbiobacterium thermophilum
11.5
-
 deamination Symbiobacterium thermophilum

Cofactor

Cofactor Comment Organism Structure
NADP+
-
 Symbiobacterium thermophilum
NADPH
-
 Symbiobacterium thermophilum

General Information

General Information Comment Organism
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