Dipeptide synthesis by an isolated adenylate‐forming domain of non‐ribosomal peptide synthetases (NRPS)

Dieckmann, Ralf; Neuhof, Torsten; Pavela-Vrančić, Maja; Döhren, Hans von

doi:10.1016/s0014-5793(01)02471-1

Cited by 13 publications

(10 citation statements)

References 18 publications

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“…This variant lacks the Ser689 attachment site to the PPant group that is involved in thioester formation and therefore would only allow the formation of the acyl adenylate and not the thioester. [26][27][28][29][30] Our current results are not able to establish if this reaction is enzyme catalyzed. [26][27][28][29][30] Our current results are not able to establish if this reaction is enzyme catalyzed.…”

Section: Zuschriftencontrasting

confidence: 76%

“…Conversion to ilepcimide 20 (79 %) was still observed with this variant, suggesting that adenylation activity alone is sufficient for amidation, presumably by nucleophilic attack onto the acyl adenylate ( Figure 2B), ar eaction which has been observed with other adenylating and phosphorylating enzymes,including NRPS and glutamine synthetase. [26][27][28][29][30] Our current results are not able to establish if this reaction is enzyme catalyzed.…”

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confidence: 76%

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Adenylation Activity of Carboxylic Acid Reductases Enables the Synthesis of Amides

et al. 2017

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Carboxylic acid reductases (CARs) catalyze the reduction of a broad range of carboxylic acids to aldehydes using the cofactors adenosine triphosphate and nicotinamide adenine dinucleotide phosphate, and have become attractive biocatalysts for organic synthesis. Mechanistic understanding of CARs was used to expand reaction scope, generating biocatalysts for amide bond formation from carboxylic acid and amine. CARs demonstrated amidation activity for various acids and amines. Optimization of reaction conditions, with respect to pH and temperature, allowed for the synthesis of the anticonvulsant ilepcimide with up to 96 % conversion. Mechanistic studies using site-directed mutagenesis suggest that, following initial enzymatic adenylation of substrates, amidation of the carboxylic acid proceeds by direct reaction of the acyl adenylate with amine nucleophiles.

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Section: Zuschriftencontrasting

confidence: 76%

contrasting

confidence: 76%

Adenylation Activity of Carboxylic Acid Reductases Enables the Synthesis of Amides

et al. 2017

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“…In fact, covalent capture of the highly reactive aminoacyl-adenylate intermediate with diffusable small molecules has been observed as a side reaction for other NRPSs. [64,78] NovL is an example for the efficient exploitation of this kind of reaction, which can obviously be encountered in many biosynthetic pathways requiring only one condensation step between two precursors of the final product. [79] It can also be compared with the reaction catalyzed by acyl-CoA ligases which belong to the same superfamily as the A domains.…”

Section: Nonlinear Nrpss (Type C)mentioning

confidence: 99%

Ways of Assembling Complex Natural Products on Modular Nonribosomal Peptide Synthetases A list of abbreviations can be found at the end of the text.

2002

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Nonribosomal peptide synthetases (NRPSs) catalyze the assembly of a large number of complex peptide natural products, many of which display therapeutically useful activity. Each cycle of chain extension is carried out by a dedicated module of the multifunctional enzymes. A module harbors all the catalytic units, which are referred to as domains, necessary for recognition, activation, covalent binding, and optionally modification of a single building block monomer, as well as for peptide-bond formation with the growing chain. A terminal domain releases the full-length peptide chain from the enzyme complex. Recent characterization of many NRPS systems revealed several examples where the sequence of the product does not directly correspond to the linear arrangement of modules and domains within the enzyme(s). It is now obvious that these systems cannot be regarded as rare exceptions of the common NRPS architecture but rather represent more complicated variations of the NRPS repertoire to increase their biosynthetic potential. In most of these cases unusual peptide structures of the products are observed, such as structures with side-chain acylation, cyclization involving the peptide backbone and/or side chains, and transfer of the peptide chain onto soluble small-molecule substrates. These findings indicate a previously unexpected higher versatility of the modules and domains in terms of both catalytic potential and interaction within the multifunctional protein templates. We propose to classify the known NRPS systems into three groups, linear NRPSs (type A), iterative NRPSs (type B), and nonlinear NRPSs (type C), according to their biosynthetic logic. Understanding the various biosynthetic strategies of NRPSs will be crucial to fully explore their potential for engineered combinatorial biosynthesis.

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“…Bacteria and fungi possess NRPSs to synthesize antibiotic peptides such as cyclosporin A, gramicidin S [7], enterobactin [31], tyrocidine [32] and acinetobactin [33]. NRPSs have multiple components which each add a single amino acid to the antibiotic peptide.…”

Section: Introductionmentioning

confidence: 99%

In silico analysis of class I adenylate-forming enzymes reveals family and group-specific conservations

et al. 2018

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Luciferases, aryl- and fatty-acyl CoA synthetases, and non-ribosomal peptide synthetase proteins belong to the class I adenylate-forming enzyme superfamily. The reaction catalyzed by the adenylate-forming enzymes is categorized by a two-step process of adenylation and thioesterification. Although all of these proteins perform a similar two-step process, each family may perform the process to yield completely different results. For example, luciferase proteins perform adenylation and oxidation to produce the green fluorescent light found in fireflies, while fatty-acyl CoA synthetases perform adenylation and thioesterification with coenzyme A to assist in metabolic processes involving fatty acids. This study aligned a total of 374 sequences belonging to the adenylate-forming superfamily. Analysis of the sequences revealed five fully conserved residues throughout all sequences, as well as 78 more residues conserved in at least 60% of sequences aligned. Conserved positions are involved in magnesium and AMP binding and maintaining enzyme structure. Also, ten conserved sequence motifs that included most of the conserved residues were identified. A phylogenetic tree was used to assign sequences into nine different groups. Finally, group entropy analysis identified novel conservations unique to each enzyme group. Common group-specific positions identified in multiple groups include positions critical to coordinating AMP and the CoA-bound product, a position that governs active site shape, and positions that help to maintain enzyme structure through hydrogen bonds and hydrophobic interactions. These positions could serve as excellent targets for future research.

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Dipeptide synthesis by an isolated adenylate‐forming domain of non‐ribosomal peptide synthetases (NRPS)

Cited by 13 publications

References 18 publications

Adenylation Activity of Carboxylic Acid Reductases Enables the Synthesis of Amides

Adenylation Activity of Carboxylic Acid Reductases Enables the Synthesis of Amides

Ways of Assembling Complex Natural Products on Modular Nonribosomal Peptide Synthetases A list of abbreviations can be found at the end of the text.

In silico analysis of class I adenylate-forming enzymes reveals family and group-specific conservations

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