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Literature summary for 6.2.1.54 extracted from

  • Luo, Y.; Du, L.
    Thiolation-enhanced substrate recognition by D-alanyl carrier protein ligase DltA from Bacillus cereus (2014), F1000Res., 3, 106 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.2.1.54

Activating Compound

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(Commentary)

Cloned (Comment) Organism
-
 Bacillus cereus

Crystallization (Commentary)

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

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

Inhibitors Comment Organism Structure
5'-O-[N-(D-alanyl)-sulfamoyl]adenosine
-
 Bacillus cereus

KM Value [mM]

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Bacillus cereus Q81G39
-
-
Bacillus cereus DSM 31 Q81G39
-
-

Purification (Commentary)

Purification (Comment) Organism
nickel affinity column chromatography and gel filtration Bacillus cereus

Substrates and Products (Substrate)

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

Synonyms Comment Organism
D-alanyl carrier protein ligase
-
 Bacillus cereus
DltA
-
 Bacillus cereus

Turnover Number [1/s]

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

Cofactor Comment Organism Structure
ATP
-
 Bacillus cereus

kcat/KM [mM/s]

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