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Literature summary extracted from

  • Murcia, N.; Lee, X.; Waridel, P.; Maspoli, A.; Imker, H.; Chai, T.; Walsh, C.; Reimmann, C.
    The Pseudomonas aeruginosa antimetabolite L-2-amino-4-methoxy-trans-3-butenoic acid (AMB) is made from glutamate and two alanine residues via a thiotemplate-linked tripeptide precursor (2015), Front. Microbiol., 6, 170 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.2.1.67 gene ambB, encoded in the amb gene cluster, genetic organization, recombinant overexpression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)/pLys Pseudomonas aeruginosa
6.2.1.68 gene ambE, encoded in the amb gene cluster, genetic organization, recombinant overexpression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)/pLys Pseudomonas aeruginosa

Protein Variants

EC Number Protein Variants Comment Organism
6.2.1.68 D644A site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely Pseudomonas aeruginosa
6.2.1.68 D644A/K1230T/S1985A site-directed mutagenesis, the mutations abolish loading of L-Glu completely Pseudomonas aeruginosa
6.2.1.68 K1230T site-directed mutagenesis, amino acid alterations in the A domain abolish loading of L-Glu entirely Pseudomonas aeruginosa
6.2.1.68 S1286A site-directed mutagenesis, the mutation abolishes loading of L-Glu almost completely Pseudomonas aeruginosa
6.2.1.68 S1819A site-directed mutagenesis, mutation of S1819 does not interfere with loading of L-Glu Pseudomonas aeruginosa
6.2.1.68 S1958A site-directed mutagenesis, mutation of the active site Ser (S1958) of the TE domain does not interfere with loading of L-Glu Pseudomonas aeruginosa

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.2.1.67 Mg2+ required Pseudomonas aeruginosa
6.2.1.68 Mg2+ required Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa
-
AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa ATCC 15692
-
AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa 1C
-
AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa PRS 101
-
AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa DSM 22644
-
AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa CIP 104116
-
AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa LMG 12228
-
AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa JCM 14847
-
AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate Pseudomonas aeruginosa
-
diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate Pseudomonas aeruginosa ATCC 15692
-
diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate Pseudomonas aeruginosa 1C
-
diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate Pseudomonas aeruginosa PRS 101
-
diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate Pseudomonas aeruginosa DSM 22644
-
diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate Pseudomonas aeruginosa CIP 104116
-
diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate Pseudomonas aeruginosa LMG 12228
-
diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate Pseudomonas aeruginosa JCM 14847
-
diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa
-
AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa ATCC 15692
-
AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa 1C
-
AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa PRS 101
-
AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa DSM 22644
-
AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa CIP 104116
-
AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa LMG 12228
-
AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein] Pseudomonas aeruginosa JCM 14847
-
AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa
-
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa ATCC 15692
-
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa 1C
-
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa PRS 101
-
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa DSM 22644
-
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa CIP 104116
-
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa LMG 12228
-
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa JCM 14847
-
AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate Pseudomonas aeruginosa
-
diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate Pseudomonas aeruginosa ATCC 15692
-
diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate Pseudomonas aeruginosa 1C
-
diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate Pseudomonas aeruginosa PRS 101
-
diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate Pseudomonas aeruginosa DSM 22644
-
diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate Pseudomonas aeruginosa CIP 104116
-
diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate Pseudomonas aeruginosa LMG 12228
-
diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate Pseudomonas aeruginosa JCM 14847
-
diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa
-
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa ATCC 15692
-
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa 1C
-
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa PRS 101
-
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa DSM 22644
-
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa CIP 104116
-
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa LMG 12228
-
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein] Pseudomonas aeruginosa JCM 14847
-
AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir

Organism

EC Number Organism UniProt Comment Textmining
6.2.1.67 Pseudomonas aeruginosa Q9I1H0
-
-
6.2.1.67 Pseudomonas aeruginosa 1C Q9I1H0
-
-
6.2.1.67 Pseudomonas aeruginosa ATCC 15692 Q9I1H0
-
-
6.2.1.67 Pseudomonas aeruginosa CIP 104116 Q9I1H0
-
-
6.2.1.67 Pseudomonas aeruginosa DSM 22644 Q9I1H0
-
-
6.2.1.67 Pseudomonas aeruginosa JCM 14847 Q9I1H0
-
-
6.2.1.67 Pseudomonas aeruginosa LMG 12228 Q9I1H0
-
-
6.2.1.67 Pseudomonas aeruginosa PRS 101 Q9I1H0
-
-
6.2.1.68 Pseudomonas aeruginosa Q9I1H3
-
-
6.2.1.68 Pseudomonas aeruginosa 1C Q9I1H3
-
-
6.2.1.68 Pseudomonas aeruginosa ATCC 15692 Q9I1H3
-
-
6.2.1.68 Pseudomonas aeruginosa CIP 104116 Q9I1H3
-
-
6.2.1.68 Pseudomonas aeruginosa DSM 22644 Q9I1H3
-
-
6.2.1.68 Pseudomonas aeruginosa JCM 14847 Q9I1H3
-
-
6.2.1.68 Pseudomonas aeruginosa LMG 12228 Q9I1H3
-
-
6.2.1.68 Pseudomonas aeruginosa PRS 101 Q9I1H3
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.2.1.67 recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3)/pLys by nickel affinity chromatography Pseudomonas aeruginosa
6.2.1.68 recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)/pLys by nickel affinity chromatography Pseudomonas aeruginosa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa ATCC 15692 AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa 1C AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa PRS 101 AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa DSM 22644 AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa CIP 104116 AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa LMG 12228 AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 (L-alanyl)adenylate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa JCM 14847 AMP + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate
-
Pseudomonas aeruginosa diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate
-
Pseudomonas aeruginosa ATCC 15692 diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate
-
Pseudomonas aeruginosa 1C diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate
-
Pseudomonas aeruginosa PRS 101 diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate
-
Pseudomonas aeruginosa DSM 22644 diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate
-
Pseudomonas aeruginosa CIP 104116 diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate
-
Pseudomonas aeruginosa LMG 12228 diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate
-
Pseudomonas aeruginosa JCM 14847 diphosphate + (L-alanyl)adenylate
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa ATCC 15692 AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa 1C AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa PRS 101 AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa DSM 22644 AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa CIP 104116 AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa LMG 12228 AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 ATP + L-glutamate + holo-[L-alanyl-carrier protein]
-
Pseudomonas aeruginosa JCM 14847 AMP + diphosphate + L-alanyl-[L-alanyl-carrier protein]
-
ir
6.2.1.67 additional information enzyme AmbE (L-glutamate-[L-glutamyl-carrier protein] ligase EC 6.2.1.68) is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB, AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa ?
-
-
6.2.1.67 additional information enzyme AmbE (L-glutamate-[L-glutamyl-carrier protein] ligase EC 6.2.1.68) is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB, AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa ATCC 15692 ?
-
-
6.2.1.67 additional information enzyme AmbE (L-glutamate-[L-glutamyl-carrier protein] ligase EC 6.2.1.68) is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB, AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa 1C ?
-
-
6.2.1.67 additional information enzyme AmbE (L-glutamate-[L-glutamyl-carrier protein] ligase EC 6.2.1.68) is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB, AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa PRS 101 ?
-
-
6.2.1.67 additional information enzyme AmbE (L-glutamate-[L-glutamyl-carrier protein] ligase EC 6.2.1.68) is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB, AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa DSM 22644 ?
-
-
6.2.1.67 additional information enzyme AmbE (L-glutamate-[L-glutamyl-carrier protein] ligase EC 6.2.1.68) is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB, AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa CIP 104116 ?
-
-
6.2.1.67 additional information enzyme AmbE (L-glutamate-[L-glutamyl-carrier protein] ligase EC 6.2.1.68) is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB, AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa LMG 12228 ?
-
-
6.2.1.67 additional information enzyme AmbE (L-glutamate-[L-glutamyl-carrier protein] ligase EC 6.2.1.68) is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB, AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa JCM 14847 ?
-
-
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa ATCC 15692 AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa 1C AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa PRS 101 AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa DSM 22644 AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa CIP 104116 AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa LMG 12228 AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 (L-glutamyl)adenylate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa JCM 14847 AMP + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate
-
Pseudomonas aeruginosa diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate
-
Pseudomonas aeruginosa ATCC 15692 diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate
-
Pseudomonas aeruginosa 1C diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate
-
Pseudomonas aeruginosa PRS 101 diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate
-
Pseudomonas aeruginosa DSM 22644 diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate
-
Pseudomonas aeruginosa CIP 104116 diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate
-
Pseudomonas aeruginosa LMG 12228 diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate
-
Pseudomonas aeruginosa JCM 14847 diphosphate + (L-glutamyl)adenylate
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa ATCC 15692 AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa 1C AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa PRS 101 AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa DSM 22644 AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa CIP 104116 AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa LMG 12228 AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 ATP + L-glutamate + holo-[L-glutamyl-carrier protein]
-
Pseudomonas aeruginosa JCM 14847 AMP + diphosphate + L-glutamyl-[L-glutamyl-carrier protein]
-
ir
6.2.1.68 additional information identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa ?
-
-
6.2.1.68 additional information identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa ATCC 15692 ?
-
-
6.2.1.68 additional information identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa 1C ?
-
-
6.2.1.68 additional information identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa PRS 101 ?
-
-
6.2.1.68 additional information identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa DSM 22644 ?
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6.2.1.68 additional information identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa CIP 104116 ?
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6.2.1.68 additional information identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa LMG 12228 ?
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6.2.1.68 additional information identification of L-Glu as the amino acid substrate activated by AmbE and loaded onto its T1 domain. The enzyme is also active with L-alanine instead of L-glutamate, but only in presence of the L-alanine-[L-alanyl-carrier protein] ligase AmbB (cf. EC 6.2.1.67), AmbB-dependent loading of L-Ala onto the T2 domain of AmbE, overview. Analysis of enzyme peptide fragment binding with substrate, peptide analysis by mass spectrometry, overview Pseudomonas aeruginosa JCM 14847 ?
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Subunits

EC Number Subunits Comment Organism
6.2.1.67 More modular structure of AmbE with domains for adenylation, thiolation, condensation, methylation, and thioester cleavage. AmbE may have an additional domain of unknown function at its N-terminus and the C domain is atypical, domain architecture, overview Pseudomonas aeruginosa
6.2.1.68 More modular structure of AmbE with domains for adenylation, thiolation, condensation, methylation, and thioester cleavage. AmbE may have an additional domain of unknown function at its N-terminus and the C domain is atypical, domain architecture, overview Pseudomonas aeruginosa

Synonyms

EC Number Synonyms Comment Organism
6.2.1.67 ambB
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Pseudomonas aeruginosa
6.2.1.67 More see also EC 6.2.1.68 Pseudomonas aeruginosa
6.2.1.67 non-ribosomal peptide synthetase
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Pseudomonas aeruginosa
6.2.1.68 ambE
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Pseudomonas aeruginosa
6.2.1.68 More see also EC 6.2.1.67 Pseudomonas aeruginosa
6.2.1.68 non-ribosomal peptide synthetase
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Pseudomonas aeruginosa

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.2.1.67 28
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assay at Pseudomonas aeruginosa
6.2.1.68 28
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assay at Pseudomonas aeruginosa

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.2.1.67 7.5
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assay at Pseudomonas aeruginosa
6.2.1.68 7.5
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assay at Pseudomonas aeruginosa

Cofactor

EC Number Cofactor Comment Organism Structure
6.2.1.67 4'-phosphopantetheine phosphopantetheinylation of recombinant apoenzyme AmbE by a purified promiscuous phosphopantetheinyl transferase Sfp at pH 7.5, 28°C Pseudomonas aeruginosa
6.2.1.67 ATP
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Pseudomonas aeruginosa
6.2.1.68 4'-phosphopantetheine phosphopantetheinylation of recombinant apoenzyme AmbE by a purified promiscuous phosphopantetheinyl transferase Sfp at pH 7.5, 28°C Pseudomonas aeruginosa
6.2.1.68 ATP
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Pseudomonas aeruginosa

General Information

EC Number General Information Comment Organism
6.2.1.67 metabolism Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoicacid (AMB)is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA), two non-ribosomal peptide synthetases (AmbB and AmbE, EC 6.2.1.67 and EC 6.2.1.68, respectively), and two iron(II)/alpha-ketoglutarate-dependent oxygenases (AmbC and AmbD). Bioinformatics analysis predicts one thiolation (T) domain for AmbB and two T domains (T1 and T2) for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. The AmbB substrate is identified to be L-alanine (L-Ala), while the T1 and T2 domains of AmbE are loaded with L-glutamate (L-Glu) and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurs only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE result in the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD are included in the assay, these peptides are no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide is found at T2, importance of flanking L-Ala residues in the precursor tripeptide Pseudomonas aeruginosa
6.2.1.67 additional information enzyme AmbB presents the typical modular structure of non-ribosomal peptide synthetases (NRPSs) Pseudomonas aeruginosa
6.2.1.67 physiological function the enzyme is involved in biosynthesis of Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoicacid (AMB), which proceeds via a precursor tripeptide. Identification of the building blocks of AMB biosynthesis and modelling, overview Pseudomonas aeruginosa
6.2.1.68 metabolism Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB)is a non-proteinogenic amino acid which is toxic for prokaryotes and eukaryotes. Production of AMB requires a five-gene cluster encoding a putative LysE-type transporter (AmbA), two non-ribosomal peptide synthetases (AmbB and AmbE, EC 6.2.1.67 and EC 6.2.1.68, respectively), and two iron(II)/alpha-ketoglutarate-dependent oxygenases (AmbC and AmbD). Bioinformatics analysis predicts one thiolation (T) domain for AmbB and two T domains (T1 and T2) for AmbE, suggesting that AMB is generated by a processing step from a precursor tripeptide assembled on a thiotemplate. The AmbB substrate is identified to be L-alanine (L-Ala), while the T1 and T2 domains of AmbE are loaded with L-glutamate (L-Glu) and L-Ala, respectively. Loading of L-Ala at T2 of AmbE occurs only in the presence of AmbB, indicative of a trans loading mechanism. In vitro assays performed with AmbB and AmbE result in the dipeptide L-Glu-L-Ala at T1 and the tripeptide L-Ala-L-Glu-L-Ala attached at T2. When AmbC and AmbD are included in the assay, these peptides are no longer detected. Instead, an L-Ala-AMB-L-Ala tripeptide is found at T2, importance of flanking L-Ala residues in the precursor tripeptide Pseudomonas aeruginosa
6.2.1.68 additional information enzyme AmbE presents the typical modular structure of non-ribosomal peptide synthetases (NRPSs) Pseudomonas aeruginosa
6.2.1.68 physiological function the enzyme is involved in biosynthesis of Pseudomonas aeruginosa toxin L-2-amino-4-methoxy-trans-3-butenoic acid (AMB), which proceeds via a precursor tripeptide. Identification of the building blocks of AMB biosynthesis and modelling, assembly of a tripeptide AmbB precursor on AmbE, overview Pseudomonas aeruginosa