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 (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 | 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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Symbiobacterium thermophilum | Q67PI3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant soluble enzyme 3.4fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Symbiobacterium thermophilum |
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 | Comment | Organism |
---|---|---|
meso-DAPDH | - |
Symbiobacterium thermophilum |
meso-diaminopimelate dehydrogenase | - |
Symbiobacterium thermophilum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Symbiobacterium thermophilum |
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 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
amination | Symbiobacterium thermophilum |
11.5 | - |
deamination | Symbiobacterium thermophilum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Symbiobacterium thermophilum | |
NADPH | - |
Symbiobacterium thermophilum |
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 |