Flavin-Linked Oxidase Catalyzes Pyrrolizine Formation of Dichloropyrrole-Containing Polyketide Extender Unit in Chlorizidine A

Mantovani, Simone M.; Moore, Bradley S.

doi:10.1021/ja409520v

Cited by 54 publications

(61 citation statements)

References 33 publications

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“…6 A and B) is in close proximity to the side chain of the catalytic lysine residue. An examination of the amino acids lining this putative pyrrole binding site in Bmp2 reveals a lack of conservation of three key sites (Y302, F306, and A345) that are otherwise strictly conserved among all pyrrolyl-S-ACP halogenases that selectively catalyze one [HrmQ (10)] or two [PltA (7), Mpy16 (8), Clz5 (9), and Pyr29 (18)] halogenations on the pyrrole ring (Fig. 6D).…”

Section: Resultsmentioning

confidence: 99%

“…In the first route, the ACP-loaded halopyrrole is extended using modular assembly lines to yield nonribosomally synthesized peptide, or polyketide natural products (e.g., 2-4, Fig. 1) (7)(8)(9)(10). During the biosynthesis of 2-4, and other related natural products, the halopyrrole carboxylic acid is transthioesterified to downstream ACPs during modular elongation reactions.…”

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confidence: 99%

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Biosynthesis of coral settlement cue tetrabromopyrrole in marine bacteria by a uniquely adapted brominase–thioesterase enzyme pair

Gamal

Agarwal

Diethelm

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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118

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Halogenated pyrroles (halopyrroles) are common chemical moieties found in bioactive bacterial natural products. The halopyrrole moieties of mono-and dihalopyrrole-containing compounds arise from a conserved mechanism in which a proline-derived pyrrolyl group bound to a carrier protein is first halogenated and then elaborated by peptidic or polyketide extensions. This paradigm is broken during the marine pseudoalteromonad bacterial biosynthesis of the coral larval settlement cue tetrabromopyrrole (1), which arises from the substitution of the proline-derived carboxylate by a bromine atom. To understand the molecular basis for decarboxylative bromination in the biosynthesis of 1, we sequenced two Pseudoalteromonas genomes and identified a conserved four-gene locus encoding the enzymes involved in its complete biosynthesis. Through total in vitro reconstitution of the biosynthesis of 1 using purified enzymes and biochemical interrogation of individual biochemical steps, we show that all four bromine atoms in 1 are installed by the action of a single flavin-dependent halogenase: Bmp2. Tetrabromination of the pyrrole induces a thioesterase-mediated offloading reaction from the carrier protein and activates the biosynthetic intermediate for decarboxylation. Insights into the tetrabrominating activity of Bmp2 were obtained from the high-resolution crystal structure of the halogenase contrasted against structurally homologous halogenase Mpy16 that forms only a dihalogenated pyrrole in marinopyrrole biosynthesis. Structure-guided mutagenesis of the proposed substrate-binding pocket of Bmp2 led to a reduction in the degree of halogenation catalyzed. Our study provides a biogenetic basis for the biosynthesis of 1 and sets a firm foundation for querying the biosynthetic potential for the production of 1 in marine (meta)genomes.natural products | biosynthesis | halogenation | enzymology

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Biosynthesis of coral settlement cue tetrabromopyrrole in marine bacteria by a uniquely adapted brominase–thioesterase enzyme pair

Gamal

Agarwal

Diethelm

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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118

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“…122 Figure 15A). [161][162][163] The peptidyl carrier protein (PCP) tethered substrates of Clz5, PltA and HalB are all derived from oxidation of proline-S-PCP intermediates in NRPS or NRPS-PKS assembly lines. Genome mining of the marine organisms responsible for the biosynthesis of brominated marine natural products has led to the identification of an interesting pyrrole Fl-Hal, Bmp2, from…”

Section: Flavin-dependent Pyrrole Halogenasesmentioning

confidence: 99%

Development of Halogenase Enzymes for Use in Synthesis

Latham

Brandenburger

Shepherd³

et al. 2017

Chem. Rev.

286

244

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Nature has evolved halogenase enzymes to regioselectively halogenate a diverse range of biosynthetic precursors, with the halogens introduced often having a profound effect on the biological activity of the resulting natural products. Synthetic endeavours to create non-natural bioactive small molecules for pharmaceutical and agrochemical applications have also arrived at a similar conclusion -halogens can dramatically improve the properties of organic molecules for selective modulation of biological targets in vivo. Consequently a high proportion of pharmaceuticals and agrochemicals on the market today possess halogens. Halogenated organic compounds are also common intermediates in synthesis and are particularly valuable in metal catalyzed cross-coupling reactions.Despite the potential utility of organohalogens, traditional non-enzymatic halogenation chemistry utilizes deleterious reagents and often lacks regiocontrol. Reliable, facile and cleaner methods for the regioselective halogenation of organic compounds are therefore essential in the development of economical and environmentally-friendly industrial processes. A potential avenue towards such methods is the use of halogenase enzymes, responsible for the biosynthesis of halogenated natural products, as biocatalysts. This review will discuss the advances towards this goal thus far, in addition to untapped potential sources of such biocatalysts and how further development of the enzymes may be focused in order to achieve the goal of industrial scale biohalogenation.

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“…8 has since been postulated as the starter unit in pyrrolomycin ( 9 ), marinopyrrole ( 10 ) and chlorizidine A ( 11 ) biosynthesis, following identification of the genes encoding enzymes homologous to those in the pyoluteorin biosynthetic pathway (Figure 3B). 18–20 …”

Section: Polyketide Starter Unitsmentioning

confidence: 99%

“…Oxidation by the enoyl-ACP-reductase, Clz3, yields the α,β-unsaturated thioester which is then reductively carboxylated by Clz4, the CCR, giving rise to the highly unusual extender unit 29 (Figure 6C). 20 …”

Section: Polyketide Extender Unitsmentioning

confidence: 99%

Recent advances in the biosynthesis of unusual polyketide synthase substrates

Ray

Moore

2016

Nat. Prod. Rep.

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Covering: 2003 to July 2015 for polyketide synthase starter units and 2012 to September 2015 for polyketide synthase extender units This highlight provides an overview of recent advances in understanding the diversity of polyketide synthase (PKS) substrate building blocks. Substrates functioning as starter units and extender units contribute significantly to the chemical complexity and structural diversity exhibited by this class of natural products. This article complements and extends upon the current comprehensive reviews that have been published on these two topics (Moore and Hertweck, Nat. Prod. Rep., 2002, 19, 70; Chan et. al., Nat. Prod. Rep., 2009, 1, 90; Wilson and Moore, Nat. Prod. Rep., 2012, 29, 72).

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Flavin-Linked Oxidase Catalyzes Pyrrolizine Formation of Dichloropyrrole-Containing Polyketide Extender Unit in Chlorizidine A

Cited by 54 publications

References 33 publications

Biosynthesis of coral settlement cue tetrabromopyrrole in marine bacteria by a uniquely adapted brominase–thioesterase enzyme pair

Biosynthesis of coral settlement cue tetrabromopyrrole in marine bacteria by a uniquely adapted brominase–thioesterase enzyme pair

Development of Halogenase Enzymes for Use in Synthesis

Recent advances in the biosynthesis of unusual polyketide synthase substrates

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