LysineN-hydroxylase andN-acetyltransferase of the aerobactin system of pColV plasmids inEscherichia coli

Heydel, P.; Plattner, Hans; Diekmann, H.

doi:10.1111/j.1574-6968.1987.tb02044.x

Cited by 8 publications

(3 citation statements)

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“…Four genes and their corresponding polypeptide products have been identified [129]. AerA, a 50 kDa protein, is a lysine-N 6 -oxidase [130]. AerB, 35 kDa, is N 6 -hydroxylysine : acetylCoA N 6 -acetyltransferase [131].…”

Section: Siderophores Containing Amino Acids Of Unusual Structuresmentioning

confidence: 99%

Peptide siderophores

1998

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Siderophores are low molecular weight iron chelators, produced by virtually all bacteria, fungi and some plants. They serve to deliver the essential element iron, barely soluble under aerobic conditions, into microbial cells. Siderophores are therefore important secondary metabolites which are very often based on amino acids and their derivatives. Biosynthesis, transport, regulation and chemical synthesis of natural siderophores and their analogues is of considerable interest for the protein and peptide chemist. This review gives an overview of the structural classes of peptidic siderophores, along with data on their biosynthesis. On a number of representative examples, strategies and schemes of their chemical synthesis are described.

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Section: Siderophores Containing Amino Acids Of Unusual Structuresmentioning

confidence: 99%

Peptide siderophores

1998

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“…The strain was maintained on Luria-Bertani agar slants with 50 pg/ml ampicillin and grown in 10-1 fermentors on a modified M56 medium as described previously (Heydel et al, 1987). Yield was 35 g wet cellsll0 1.…”

Section: Bacterial Strain and Cultivationmentioning

confidence: 99%

FAD and substrate analogs as probes for lysine N⁶‐hydroxylase from Escherichia coli EN 222

Macheroux

Plattner

Romaguera

et al. 1993

European Journal of Biochemistry

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Lysine N6-hydroxylase catalyzes the hydroxylation of the N-terminal amino function of L-lysine at the expense of NADPH and molecular oxygen. The enzyme also requires FAD for its catalytic activity. Unlike other flavoprotein monooxygenases, binding of FAD is rather weak with a Kd of 30 pM at 4°C. The spectral properties of FAD bound to lysine W-hydroxylase are very similar to free oxidized FAD. In the absence of substrate, the enzyme has an NADPH oxidase activity which results in the generation of hydrogen peroxide. With increasing concentration of L-lysine, the NADPH oxidase activity is enhanced up to 10-fold and the generation of hydrogen peroxide decreases. At the same time, the substrate is hydroxylated. K , values for L-lysine and FAD were determined as 105 pM and 0.7 pM, respectively. Utilizing FAD analogs, we could demonstrate that L-lysine exerts its effector role mostly on the reductive half reaction of the overall catalytic cycle. Prolonged incubation of the enzyme with either 8-chloro-or 8-fluoro-FAD gave rise to a covalently attached flavin which is formed as a result of the nucleophilic attack of a thiolate on the 8-position of the flavin. Several lines of evidence indicate that the reaction takes place in the FAD binding site of the protein. The substrate specificity was investigated using amino acids with various lengths of side chain. L-Lysine and derivatives with similar side chain length are hydroxylated by lysine Whydroxylase. Ornithine, the lower homolog of lysine, was not hydroxylated and did not affect the NADPH oxidase activity of the enzyme. On the other hand, homolysine accelerated the rate of NADPH oxidation but was not hydroxylated. Additional requirements for efficient hydroxylation were also investigated using a variety of substrate analogs. From these studies a schematic structure of the active site of the enzyme was deduced. Sequence comparison of the FAD binding site of various flavoproteins revealed possible factors for weak binding of the cofactor in the case of lysine W-hydroxylase.

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“…An expanded manual analysis of the pfb gene cluster helps explain this unexpected result. The amino acid sequence of PfbN exhibits homology to N-acetyl transferases implicated in the acetylation of Lys and Orn in other siderophores. , N5-acetyl-N5-OH-Orn is known to be synthesized prior to adenylation; thus further development of predictor tools is needed to distinguish this substrate from OH-Orn. The mechanism of N5-OH-Orn cyclization is as yet unknown; however in consideration of the mechanism of NRPS peptide bond formation cyclization must occur after incorporation of OH-Orn, making prediction through bioinformatics tools difficult.…”

Section: Resultsmentioning

confidence: 99%

Ambiguity of NRPS Structure Predictions: Four Bidentate Chelating Groups in the Siderophore Pacifibactin

Hardy

Butler

2019

J. Nat. Prod.

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Identified through a bioinformatics approach, a nonribosomal peptide synthetase gene cluster in Alcanivorax pacificus encodes the biosynthesis of the new siderophore pacifibactin. The structure of pacifibactin differs markedly from the bioinformatic prediction and contains four bidentate metal chelation sites, atypical for siderophores. Genome mining and structural characterization of pacifibactin is reported herein, as well as characterization of pacifibactin variants accessible due to a lack of adenylation domain fidelity during biosynthesis. A spectrophotometric titration of pacifibactin with Fe(III) and 13 C NMR spectroscopy of the Ga(III)-pacifibactin complex establish 1:1 metal:pacifibactin coordination and reveal which of the bidentate binding groups are coordinated to the metal. The photoreaction of Fe(III)-pacifibactin, resulting from Fe(III) coordination of the βhydroxyaspartic acid ligands, is reported.

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LysineN-hydroxylase andN-acetyltransferase of the aerobactin system of pColV plasmids inEscherichia coli

Cited by 8 publications

References 13 publications

Peptide siderophores

Peptide siderophores

FAD and substrate analogs as probes for lysine N⁶‐hydroxylase from Escherichia coli EN 222

Ambiguity of NRPS Structure Predictions: Four Bidentate Chelating Groups in the Siderophore Pacifibactin

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LysineN-hydroxylase andN-acetyltransferase of the aerobactin system of pColV plasmids inEscherichia coli

Cited by 8 publications

References 13 publications

Peptide siderophores

Peptide siderophores

FAD and substrate analogs as probes for lysine N6‐hydroxylase from Escherichia coli EN 222

Ambiguity of NRPS Structure Predictions: Four Bidentate Chelating Groups in the Siderophore Pacifibactin

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FAD and substrate analogs as probes for lysine N⁶‐hydroxylase from Escherichia coli EN 222