Activating Compound | Comment | Organism | Structure |
---|---|---|---|
coenzyme A | CoA can mimic D-alanyl carrier protein DltC. The resulting Michaelis constants in the presence of saturating CoA for both ATP and D-alanine are reduced more than 10fold as compared to the values obtained in the absence of CoA. The presence of CoA also made DltA about 100fold more selective on D-alanine over L-alanine | Bacillus cereus |
Cloned (Comment) | Organism |
---|---|
- |
Bacillus cereus |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, using 0.1 M MgCl2, 0.5 M KCl, 16% (w/v) polyethylene glycol 3350 and 0.05 M HEPES-NaOH buffer at pH 7.2 | Bacillus cereus |
substrate-free form, to 1.9 A resolution, space group P21 | Bacillus cereus |
Protein Variants | Comment | Organism |
---|---|---|
C269A | the mutant shows strongly reduced activity compared to the wild type enzyme | Bacillus cereus |
C269A | mutant displays a relaxed preference for D-alanine over L-alanine | Bacillus cereus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5'-O-[N-(D-alanyl)-sulfamoyl]adenosine | - |
Bacillus cereus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
D-alanine | wild-type, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.03 | - |
D-alanine | wild type enzyme, at pH 7.2 and 37°C | Bacillus cereus | |
0.03 | - |
D-alanine | wild-type, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.109 | - |
L-alanine | wild-type, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.46 | - |
ATP | wild type enzyme, at pH 7.2 and 37°C | Bacillus cereus | |
0.5 | - |
D-alanine | mutant C269A, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.5 | - |
D-alanine | mutant enzyme C269A, at pH 7.2 and 37°C | Bacillus cereus | |
1 | - |
D-alanine | mutant C269A, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
8.8 | - |
L-alanine | mutant C269A, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
8.8 | - |
L-alanine | mutant enzyme C269A, at pH 7.2 and 37°C | Bacillus cereus | |
20 | - |
L-alanine | mutant C269A, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
48 | - |
L-alanine | wild-type, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
109 | - |
L-alanine | wild type enzyme, at pH 7.2 and 37°C | Bacillus cereus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + D-alanine + poly(ribitol phosphate) | Bacillus cereus | - |
AMP + diphosphate + O-D-alanyl-poly(ribitol phosphate) | - |
? | |
ATP + D-alanine + poly(ribitol phosphate) | Bacillus cereus DSM 31 | - |
AMP + diphosphate + O-D-alanyl-poly(ribitol phosphate) | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus cereus | Q81G39 | - |
- |
Bacillus cereus DSM 31 | Q81G39 | - |
- |
Purification (Comment) | Organism |
---|---|
nickel affinity column chromatography and gel filtration | Bacillus cereus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(D-alanyl)adenylate + holo-[D-alanyl-carrier protein] | - |
Bacillus cereus | AMP + D-alanyl-[D-alanyl-carrier protein] | - |
? | |
(D-alanyl)adenylate + holo-[D-alanyl-carrier protein] | - |
Bacillus cereus DSM 31 | AMP + D-alanyl-[D-alanyl-carrier protein] | - |
? | |
ATP + D-alanine | - |
Bacillus cereus | (D-alanyl)adenylate + diphosphate | - |
? | |
ATP + D-alanine | - |
Bacillus cereus DSM 31 | (D-alanyl)adenylate + diphosphate | - |
? | |
ATP + D-alanine + holo-[D-alaninyl-carrier protein] | - |
Bacillus cereus | AMP + diphosphate + D-alanyl-[D-alanyl-carrier protein] | - |
? | |
ATP + D-alanine + holo-[D-alaninyl-carrier protein] | - |
Bacillus cereus DSM 31 | AMP + diphosphate + D-alanyl-[D-alanyl-carrier protein] | - |
? | |
ATP + D-alanine + poly(ribitol phosphate) | - |
Bacillus cereus | AMP + diphosphate + O-D-alanyl-poly(ribitol phosphate) | - |
? | |
ATP + D-alanine + poly(ribitol phosphate) | - |
Bacillus cereus DSM 31 | AMP + diphosphate + O-D-alanyl-poly(ribitol phosphate) | - |
? | |
ATP + L-alanine + holo-[D-alaninyl-carrier protein] | - |
Bacillus cereus | AMP + diphosphate + L-alanyl-[D-alanyl-carrier protein] | - |
? | |
ATP + L-alanine + poly(ribitol phosphate) | low activity | Bacillus cereus | AMP + diphosphate + O-L-alanyl-poly(ribitol phosphate) | - |
? | |
additional information | the ATP and D-alanine substrates of the adenylation reaction are incorporated when the DltA enzyme cycles through its thiolation conformation | Bacillus cereus | ? | - |
? | |
additional information | the ATP and D-alanine substrates of the adenylation reaction are incorporated when the DltA enzyme cycles through its thiolation conformation | Bacillus cereus DSM 31 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
D-alanyl carrier protein ligase | - |
Bacillus cereus |
DltA | - |
Bacillus cereus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.14 | - |
L-alanine | wild-type, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.18 | - |
D-alanine | wild type enzyme, at pH 7.2 and 37°C | Bacillus cereus | |
0.18 | - |
D-alanine | wild-type, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.18 | - |
D-alanine | wild-type, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.24 | - |
ATP | wild type enzyme, at pH 7.2 and 37°C | Bacillus cereus | |
0.26 | - |
D-alanine | mutant C269A, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.26 | - |
D-alanine | mutant enzyme C269A, at pH 7.2 and 37°C | Bacillus cereus | |
0.28 | - |
L-alanine | mutant C269A, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.28 | - |
L-alanine | mutant enzyme C269A, at pH 7.2 and 37°C | Bacillus cereus | |
0.32 | - |
L-alanine | wild type enzyme, at pH 7.2 and 37°C | Bacillus cereus | |
0.32 | - |
L-alanine | wild-type, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.33 | - |
D-alanine | mutant C269A, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.41 | - |
L-alanine | mutant C269A, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Bacillus cereus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
L-alanine | wild type enzyme, at pH 7.2 and 37°C | Bacillus cereus | |
0.003 | - |
L-alanine | wild-type, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.003 | - |
L-alanine | wild-type, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.02 | - |
L-alanine | mutant C269A, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.03 | - |
L-alanine | mutant C269A, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.03 | - |
L-alanine | mutant enzyme C269A, at pH 7.2 and 37°C | Bacillus cereus | |
0.05 | - |
ATP | wild type enzyme, at pH 7.2 and 37°C | Bacillus cereus | |
0.33 | - |
D-alanine | mutant C269A, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.53 | - |
D-alanine | mutant C269A, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
0.53 | - |
D-alanine | mutant enzyme C269A, at pH 7.2 and 37°C | Bacillus cereus | |
6.05 | - |
D-alanine | wild type enzyme, at pH 7.2 and 37°C | Bacillus cereus | |
6.1 | - |
D-alanine | wild-type, diphosphate assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus | |
9.2 | - |
D-alanine | wild-type, thiol assay, presence of coenzyme A, pH 7.2, temperature not specified in the publication | Bacillus cereus |