Using the pimeloyl-CoA synthetase adenylation fold to synthesize fatty acid thioesters

Abstract

Biotin is an essential vitamin in plants and mammals functioning as the carbon dioxide carrier within central lipid metabolism. Bacterial pimeloyl-CoA synthetase (BioW) acts as a highly specific substrate selection gate ensuring the integrity of the carbon chain in biotin synthesis.BioW catalyses the condensation of pimelic acid (C7 dicarboxylic acid) with CoASH in an ATP dependent manner to form pimeloyl-CoA, the first dedicated biotin building block.Multiple structures of Bacillus subtilis BioW together capture all three substrates as well as the intermediate pimeloyl-adenylate and product pyrophosphate (PP i ), and indicates that the enzyme uses an internal ruler to select the correct dicarboxylic acid substrate. Both the catalytic mechanism and the surprising stability of the adenylate intermediate were rationalized through site-directed mutagenesis. Building on this understanding, BioW was engineered to synthesise high value heptanoyl (C7) and octanoyl (C8) mono carboxylic acidCoA and C8 dicarboxylic-CoA products, highlighting the enzyme's synthetic potential.2

The carbon chain-selective adenylation enzyme TamA: the missing link between fatty acid and pyrrole natural product biosynthesis

“…To determine if an acyl-adenylate was non-covalently bound in the as-purified TamA, we employed native mass spectrometry that we recently used to observe a BioW/pimeloyl-adenylate complex. 29 Fig. 3c shows the native MS of TamA directly after purification with two species observed over three charge states (18+ to 16+) with nominal masses of 75 800 and 75 280 Da.…”

An Unusual Fatty Acyl:Adenylate Ligase (FAAL)–Acyl Carrier Protein (ACP) Didomain in Ambruticin Biosynthesis

“…The members of the ANL class I superfamily are divided into acyl‐ and aryl CoA synthetases, adenylation (A) domains of nonribosomal peptide synthetases (NRPSs), and firefly luciferases. Recently, the pimeloyl‐CoA synthetase BioW was discovered to represent a novel type of ANL enzyme that is able to carry out adenylation, thioesterification, and proofreading …”

Targeting adenylate-forming enzymes with designed sulfonyladenosine inhibitors

“…In a striking example of parallel evolution, at least nine distinct protein classes comprising eight different structural folds are known to catalyze adenylation reactions: Class I aminoacyl-tRNA synthetases [1], Class II aminoacyl-tRNA synthetases [1], the ANL ( a cyl-CoA synthetase, n on-ribosomal peptide synthetase (NRPS), l uciferase) family [35], ubiquitin-family E1 activating enzymes [4, 36], biotin protein ligases (which share the same fold as Class II aminoacyl-tRNA synthetases but use a distinct active-site architecture) [37, 38], N -type ATP pyrophosphatases, YrdC-like carbamoyltransferases [9], NRPS-independent siderophore synthetases [24], and the recently described BioW acyl-CoA synthetases [39, 40] (Fig. 2 and Table 1).…”

“…Recently, a ninth class of adenylate-forming enzymes was discovered. The pimeloyl-CoA synthetase BioW comprises a new catalytic fold for adenylate-forming enzymes, with the active site sandwiched between a small N-terminal domain and a larger C-terminal domain [39, 40] (Fig. 2h).…”