Cofactor-Independent Pinacolase Directs Non-Diels-Alderase Biogenesis of the Brevianamides

Ye, Ying; Du, Lan; Zhang, Qizhou; Newmister, Sean A.; Zhang, Wei; Mu, Shuai; Minami, Atsushi; McCauley, Morgan; Alegre‐Requena, Juan V.; Fraley, Amy E.; Adrover-Castellano, Maria L.; Carney, Nolan; Shende, Vikram V.; Oikawa, Hideaki; Kato, Hiromitsu; Tsukamoto, Sachiko; Paton, Robert S.; Williams, Robert M.; Sherman, David H.; Li, Shengying

doi:10.26434/chemrxiv.9122009

Cited by 9 publications

(16 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…7B) [50]. Therefore, the preponderance of 17 over 18 is likely due to intrinsic energy barriers in the cycloaddition reaction (nonenzyme catalyzed) [52]. Through the utilization of frontier molecular orbital theory, it was predicted that, for the energy levels of a relatively electron-rich diene (such as that in the DKP) to effectively interact with a dienophile, powerful electron-withdrawing groups would need to be present in the dienophile.…”

Section: )mentioning

confidence: 99%

“…An alternative mechanism for the formation of the bicyclo[2.2.2]diazaoctane ring was discovered for the biosynthesis of brevianamide A [52]. While the previously discussed MKP pathways involved formation of the bicyclic core prior to spirocycle formation, brevianamide production employs a reversed sequence of biosynthetic steps where the oxidized indole intermediate is formed prior to a subsequent DKP oxidation and cyclization.…”

Section: Enzymologymentioning

confidence: 99%

“…Since the prenyl group is an isolated electron‐neutral vinyl group, one would not expect the [4 + 2] cycloaddition in DKPs to be spontaneous without catalysis, but biomimetic syntheses [53] have shown that the azadiene undergoes a spontaneous IMDA to generate a mixture of brevianamides with enantiomorphic bicyclo[2.2.2] ring systems. While spontaneous, diastereocontrol of the IMDA may be impacted by upstream transformations such as the early‐stage semipinacol rearrangement to generate the 3‐spiro‐ψ‐indoxyl species [52].…”

Section: Enantioselective Diels–alderasesmentioning

confidence: 99%

“…This can either cyclize to form brevianamide E ( 45 ) or undergo a semipinacol rearrangement to form an indoxyl intermediate. Oxidation to the azadiene would provide the proper substrate for the IMDA [50,52]. The semipinacol rearrangement is proposed to proceed through general base catalysis to generate the indoxyl [52].…”

Section: Flavin Monooxygenases and Spirocycle Formationmentioning

confidence: 99%

“…Oxidation to the azadiene would provide the proper substrate for the IMDA [50,52]. The semipinacol rearrangement is proposed to proceed through general base catalysis to generate the indoxyl [52]. In comparison, paraherquamide biosynthesis utilizes general acid catalysis to generate an oxindole moiety [61], demonstrating that these systems have evolved divergent mechanisms for spirocycle formation.…”

Section: Flavin Monooxygenases and Spirocycle Formationmentioning

confidence: 99%

See 4 more Smart Citations

Enzyme evolution in fungal indole alkaloid biosynthesis

Fraley

Sherman

2020

The FEBS Journal

Self Cite

View full text Add to dashboard Cite

The class of fungal indole alkaloids containing the bicyclo[2.2.2]diazaoctane ring is comprised of diverse molecules that display a range of biological activities. While much interest has been garnered due to their therapeutic potential, this class of molecules also displays unique chemical functionality, making them intriguing synthetic targets. Many elegant and intricate total syntheses have been developed to generate these alkaloids, but the selectivity required to produce them in high yield presents great barriers. Alternatively, if we can understand the molecular mechanisms behind how fungi make these complex molecules, we can leverage the power of nature to perform these chemical transformations. Here, we describe the various studies regarding the evolutionary development of enzymes involved in fungal indole alkaloid biosynthesis.

show abstract

Section: )mentioning

confidence: 99%

Section: Enzymologymentioning

confidence: 99%

Section: Enantioselective Diels–alderasesmentioning

confidence: 99%

Section: Flavin Monooxygenases and Spirocycle Formationmentioning

confidence: 99%

Section: Flavin Monooxygenases and Spirocycle Formationmentioning

confidence: 99%

See 3 more Smart Citations

Enzyme evolution in fungal indole alkaloid biosynthesis

Fraley

Sherman

2020

The FEBS Journal

Self Cite

View full text Add to dashboard Cite

show abstract

Molecular Dynamics Simulations Guide Chimeragenesis and Engineered Control of Chemoselectivity in Diketopiperazine Dimerases

et al. 2023

View full text Add to dashboard Cite

In the biosynthesis of the tryptophan‐linked dimeric diketopiperazines (DKPs), cytochromes P450 selectively couple DKP monomers to generate a variety of intricate and isomeric frameworks. To determine the molecular basis for selectivity of these biocatalysts we obtained a high‐resolution crystal structure of selective Csp2−N bond forming dimerase, AspB. Overlay of the AspB structure onto C−C and C−N bond forming homolog NzeB revealed no significant structural variance to explain their divergent chemoselectivities. Molecular dynamics (MD) simulations identified a region of NzeB with increased conformational flexibility relative to AspB, and interchange of this region along with a single active site mutation led to a variant that catalyzes exclusive C−N bond formation. MD simulations also suggest that intermolecular C−C or C−N bond formation results from a change in mechanism, supported experimentally through use of a substrate mimic.

show abstract

Coordination of Polyketide Release and Multiple Detoxification Pathways for Tolerable Production of Fungal Mycotoxins

Cao

et al. 2022

Angewandte Chemie

View full text Add to dashboard Cite

Efficient biosynthesis of microbial bioactive natural products (NPs) is beneficial for the survival of producers, while self-protection is necessary to avoid self-harm resulting from over-accumulation of NPs. The underlying mechanisms for the effective but tolerable production of bioactive NPs are not well understood. Herein, in the biosynthesis of two fungal polyketide mycotoxins aurovertin E (1) and asteltoxin, we show that the cyclases in the gene clusters promote the release of the polyketide backbone, and reveal that a signal peptide is crucial for their subcellular localization and full activity. Meanwhile, the fungus adopts enzymatic acetylation as the major detoxification pathway of 1. If intermediates are over-produced, the non-enzymatic shunt pathways work as salvage pathways to avoid excessive accumulation of the toxic metabolites for selfprotection. These findings provided new insight into the interplay of efficient backbone release and multiple detoxification strategies for the production of fungal bioactive NPs.Microbial natural products (NPs) have diverse bioactivities. They protect producers from predators, antagonize the growth of other species or hosts, and help to compete for the ecological niche. Some bioactive NPs, such as antibiotics and mycotoxins, are important to compete against other organisms within the same ecological space. Therefore, they are usually produced abundantly to maximize the competitive capacity of their producers. [1] It has been suggested that the efficient release of NP backbones from the enzymatic assembly lines has contributed significantly to the NP yields. [2] The main types of NPs in nature, such as polyketides (PKs), non-ribosomal peptides (NRPs), ribosomally synthesized and post-translationally modified peptides (RiPPs), and terpenes, are released through cyclization, hydrolysis and reduction. [3] These reactions can be catalyzed by assembly enzyme-fused domains or stand-alone enzymes, such as condensation domains, thioesterases, reductases and product templates. [4] In bacteria, PKs are also liberated by the stand-alone small cyclases, [5] which belong to the nuclear transport factor 2 (NTF2)-like superfamily SnoaL-2 proteins. These proteins have also been shown to act as postline tailoring enzymes, catalyzing highly selective [6+4] cycloaddition and polyether formation. [6] In fungi, an NTF2like superfamily protein has been reported to catalyze semipinacol rearrangement. [7] But their ability to act as cyclases is poorly characterized. Meanwhile, NP producers, have evolved exquisite strategies to avoid self-harm from bioactive NPs, [8] in particular if producers also contain the conserved target of NPs. These detoxification strategies include export by transporters or subcellular trafficking, neutralization by duplication of self-resistance target proteins, and NP inactivation by enzyme-catalyzed chemical modifications. [1b, 8, 9] However, the regulatory coordination between the backbone release and detoxification in fungi for the physiologically tolerable pr...

show abstract

Cofactor-Independent Pinacolase Directs Non-Diels-Alderase Biogenesis of the Brevianamides

Cited by 9 publications

References 0 publications

Enzyme evolution in fungal indole alkaloid biosynthesis

Enzyme evolution in fungal indole alkaloid biosynthesis

Molecular Dynamics Simulations Guide Chimeragenesis and Engineered Control of Chemoselectivity in Diketopiperazine Dimerases

Coordination of Polyketide Release and Multiple Detoxification Pathways for Tolerable Production of Fungal Mycotoxins

Contact Info

Product

Resources

About