Thiazoline-Specific Amidohydrolase PurAH Is the Gatekeeper of Bottromycin Biosynthesis

Sikandar, Asfandyar; Franz, Laura; Melse, Okke; Antes, Iris; Koehnke, Jesko

doi:10.1021/jacs.8b12231

Cited by 24 publications

(31 citation statements)

References 33 publications

(77 reference statements)

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“…4,5,14,15,[6][7][8][9][10][11][12][13] During biosynthesis of complex RiPPs, when multiple enzymes capable of differentially modifying their substrate act together, a multitude of products can often form, and yet usually the biosynthetic pathway manages to produce a single natural product. Molecular mechanisms controlling the integrity of RiPP biosynthesis are only beginning to be elucidated, 2,[16][17][18][19][20][21][22][23] and many details remain unclear, especially in the cases where enzymes can apparently compete over the substrate. For example, during biosynthesis of some thiopeptides, Ser and Thr residues in the precursor peptide CP are selectively modified to either oxazoline/oxazole or dehydroamino acids by cyclodehydratase/dehydrogenase and dehydratase enzymes, and the basis for such a cooperative action despite the potential for competition has not yet been firmly established.…”

Section: Main Textmentioning

confidence: 99%

Promiscuous Enzymes Cooperate at the Substrate Level En Route to Lactazole A

Vinogradov

Shimomura²,

Kano

et al. 2020

J. Am. Chem. Soc.

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Enzymes involved in ribosomally synthesized and post-translationally modified peptide (RiPP) biosynthesis often have relaxed specificity profiles and are able to modify diverse substrates. When several such enzymes act together during precursor peptide maturation, a multitude of products can form, and yet usually, the biosynthesis converges on a single natural product. For the most part, the mechanisms controlling the integrity of RiPP assembly remain elusive. Here, we investigate biosynthesis of lactazole A, a model thiopeptide produced by five promiscuous enzymes from a ribosomal precursor peptide.Using our in vitro thiopeptide production (FIT-Laz) system, we determine the order of biosynthetic events at the individual modification level, and supplement this study with substrate scope analysis for participating enzymes. Combined, our results reveal a dynamic thiopeptide assembly process with multiple points of kinetic control, intertwined enzymatic action, and the overall substrate-level cooperation between the enzymes. This work advances our understanding of RiPP biosynthesis processes and facilitates thiopeptide bioengineering..

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Section: Main Textmentioning

confidence: 99%

Promiscuous Enzymes Cooperate at the Substrate Level En Route to Lactazole A

Vinogradov

Shimomura²,

Kano

et al. 2020

J. Am. Chem. Soc.

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“…When alone, RimO has a reversible action. However, in concert with ATP‐dependent factor RimOB (YcaO), the reaction becomes irreversible (Sikandar et al , 2019), witnessing yet another example of a protein that acts as a Maxwell's demon [see Sherrington (1940) p. 78 for an ‘animist’ attempt to use the concept as a physicochemical metaphor of the cell's life] needed to discriminate the relevant peptide against similar ones, while asking for compulsory energy dissipation to be reset to their original state (Boel et al , 2019).…”

Section: Adomet‐dependent Methylations Of the Translation Machinerymentioning

confidence: 99%

One‐carbon metabolism, folate, zinc and translation

Danchin

Sekowska

You

2020

Microbial Biotechnology

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The translation process, central to life, is tightly connected to the one-carbon (1-C) metabolism via a plethora of macromolecule modifications and specific effectors. Using manual genome annotations and putting together a variety of experimental studies, we explore here the possible reasons of this critical interaction, likely to have originated during the earliest steps of the birth of the first cells.Methionine, S-adenosylmethionine and tetrahydrofolate dominate this interaction. Yet, 1-C metabolism is unlikely to be a simple frozen accident of primaeval conditions. Reactive 1-C species (ROCS) are buffered by the translation machinery in a way tightly associated with the metabolism of iron-sulfur clusters, zinc and potassium availability, possibly coupling carbon metabolism to nitrogen metabolism. In this process, the highly modified position 34 of tRNA molecules plays a critical role. Overall, this metabolic integration may serve both as a protection against the deleterious formation of excess carbon under various growth transitions or environmental unbalanced conditions and as a regulator of zinc homeostasis, while regulating input of prosthetic groups into nascent proteins. This knowledge should be taken into account in metabolic engineering.

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“…Finally, although we show that the methyltransferase LahS B acts independently of LahM activity, we cannot exclude the possibility that LahM1/M2 substrate recognition and therefore activity are predicated on the presence of other PTMs. A strict sequence of PTM installation has been observed previously for some RiPP biosynthetic pathways such as the thiopeptides, select lanthipeptides, microcin C7, and the bottromycins . However, in other systems such as the cyanobactins, the biosynthetic enzymes have demonstrated much plasticity and can act in a variety of orders .…”

Section: Resultsmentioning

confidence: 64%

Characterization of a Dehydratase and Methyltransferase in the Biosynthesis of Ribosomally Synthesized and Post‐translationally Modified Peptides in Lachnospiraceae

et al. 2019

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As a result of the exponential increase in genomic data, discovery of novel ribosomally synthesized and post‐translationally modified peptide natural products (RiPPs) has progressed rapidly in the past decade. The lanthipeptides are a major subset of RiPPs. Through genome mining we identified a novel lanthipeptide biosynthetic gene cluster (lah) from Lachnospiraceae bacterium C6A11, an anaerobic bacterium that is a member of the human microbiota and which is implicated in the development of host disease states such as type 2 diabetes and resistance to Clostridium difficile colonization. The lah cluster encodes at least seven putative precursor peptides and multiple post‐translational modification (PTM) enzymes. Two unusual class II lanthipeptide synthetases LahM1/M2 and a substrate‐tolerant S‐adenosyl‐l‐methionine (SAM)‐dependent methyltransferase LahSB are biochemically characterized in this study. We also present the crystal structure of LahSB in complex with product S‐adenosylhomocysteine. This study sets the stage for further exploration of the final products of the lah pathway as well as their potential physiological functions in human/animal gut microbiota.

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Thiazoline-Specific Amidohydrolase PurAH Is the Gatekeeper of Bottromycin Biosynthesis

Abstract: The ribosomally synthesized and post-translationally modified peptide (RiPP) bottromycin A2 possesses potent antimicrobial activity. Its biosynthesis involves the enzymatic formation of a macroamidine, a process previously suggested to require the concerted efforts of a YcaO enzyme

Cited by 24 publications

References 33 publications

Promiscuous Enzymes Cooperate at the Substrate Level En Route to Lactazole A

Promiscuous Enzymes Cooperate at the Substrate Level En Route to Lactazole A

One‐carbon metabolism, folate, zinc and translation

Characterization of a Dehydratase and Methyltransferase in the Biosynthesis of Ribosomally Synthesized and Post‐translationally Modified Peptides in Lachnospiraceae

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