Dichlorinated and Brominated Rugulovasines, Ergot Alkaloids Produced by Talaromyces wortmannii

Medeiros, Lívia Soman de; Silva, José Vinicius da; Abreu, Lucas M.; Pfenning, Ludwig H.; Silva, Carolina Lúcia; Thomasi, Sérgio Scherrer; Venâncio, Tiago; Pée, Karl‐Heinz van; Nielsen, Kristian Fog; Rodrigues‐Filho, Edson

doi:10.3390/molecules200917627

Cited by 11 publications

(9 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The m/z 269.1 analytes of the easH-and easQ-transformed N. fumigata easA knockout strain and P. biforme NRRL 885 fragmented similarly ( Fig. 4) and in a manner consistent with the high-resolution-fragment ions obtained from rugulovasine A and B in previous work (37). Whereas rugulovasines were easily detected by LC-MS, they were not detected by HPLC with fluorescence detection (monitoring at excitation/emission wavelengths of 272 nm/372 nm or 310 nm/410 nm, respectively), which contrasts with other ergot alkaloids.…”

Section: Resultssupporting

confidence: 84%

Ergot Alkaloid Synthesis Capacity of Penicillium camemberti

Fabian

Maust

Panaccione

2018

Appl Environ Microbiol

View full text Add to dashboard Cite

Ergot alkaloids are specialized fungal metabolites with potent biological activities. They are encoded by well-characterized gene clusters in the genomes of producing fungi. plays a major role in the ripening of Brie and Camembert cheeses. The genome contains a cluster of five genes shown in other fungi to be required for synthesis of the important ergot alkaloid intermediate chanoclavine-I aldehyde and two additional genes ( and ) that may control modification of chanoclavine-I aldehyde into other ergot alkaloids. We analyzed samples of Brie and Camembert cheeses, as well as cultures of, and did not detect chanoclavine-I aldehyde or its derivatives. To create a functioning facsimile of the cluster, we expressed and in a chanoclavine-I aldehyde-accumulating knockout mutant of The-engineered strain accumulated a pair of compounds of 269.1288 in positive-mode liquid chromatography-mass spectrometry (LC-MS). The analytes fragmented in a manner typical of the stereoisomeric ergot alkaloids rugulovasine A and B, and the related rugulovasine producer accumulated the same isomeric pair of analytes. The genes were transcribed in culture, but comparison of the cluster with the functional cluster from indicated 11 polymorphisms. Whereas other genes functioned when expressed in, did not restore ergot alkaloids when expressed in an mutant. The data indicate that formerly had the capacity to produce the ergot alkaloids rugulovasine A and B. The presence of ergot alkaloid synthesis genes in the genome of is significant, because the fungus is widely consumed in Brie and Camembert cheeses. Our results show that, although the fungus has several functional genes from the ergot alkaloid pathway, it produces only an early pathway intermediate in culture and does not produce ergot alkaloids in cheese., a close relative of , contains a similar but fully functional set of ergot alkaloid synthesis genes and produces ergot alkaloids chanoclavine-I, chanoclavine-I aldehyde, and rugulovasine A and B. Our reconstruction ofthe pathway in the model fungus indicated that formerly had the capacity to produce these same ergot alkaloids. Neither nor produced ergot alkaloids in cheese, indicating that nutritionally driven gene regulation prevents these fungi from producing ergot alkaloids in a dairy environment.

show abstract

Section: Resultssupporting

confidence: 84%

Ergot Alkaloid Synthesis Capacity of Penicillium camemberti

Fabian

Maust

Panaccione

2018

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…Our LC-MS data indicate that P. biforme and the easH/easQ-transformed easA ko strain of N. fumigata are capable of producing both rugulovasine A and B ( Figure 6). This is evident as the fragmentation spectra for both rugulovasine A and B match the fragmentation spectra of rugulovasines from previous studies (De Medeiros et al, 2015). However, the chlorinated and brominated forms of rugulovasine were not detected when analyzed on LC-MS based on the absence of analytes of the predicted masses established in analyses by De Medeiros et al (2015).…”

Section: Rugulovasinessupporting

confidence: 83%

“…Ergot alkaloids are great chemical structures to undergo biohalogenation as they contain nitrogen in their chemical structure (Gribble, 2008). Thus, it there are also naturally-occurring chlorinated and brominated forms of rugulovasine (De Medeiros et al, 2015).…”

Section: Rugulovasinesmentioning

confidence: 99%

Ergot Alkaloid Synthetic Capacity of Penicillium camemberti

Fabian¹

View full text Add to dashboard Cite

Penicillium camemberti plays a major role in the ripening process of brie and camembert type cheeses. Investigation of the recently sequenced P. camemberti genome revealed the presence of a cluster of five genes previously shown to be required for ergot alkaloid synthesis in other fungi. Clustered with the five ergot alkaloid synthesis genes (aka eas genes) were two additional genes with that had the apparent capacity to encode enzymes involved in secondary metabolism. We analyzed samples of brie and camembert cheeses as well as cultures of P. camemberti grown under different conditions and were not able to detect any known ergot alkaloids, indicating the P. camemberti eas genes were either not expressed or encoded non-functional enzymes. We used a heterologous expression strategy to investigate the theoretical biosynthetic capacity of P. camemberti. Based on studies with the related ergot alkaloid-producing fungus Neosartorya fumigata (syn. Aspergillus fumigatus), the five known eas genes found clustered in the P. camemberti genome should give the fungus the capacity to produce the ergot alkaloid chanoclavine-I aldehyde. We used a chanoclavine-I aldehyde-accumulating mutant of N. fumigata as a recipient strain in which to express the two uncharacterized P. camemberti eas cluster genes (named easH and easQ) to create a functioning facsimile of the P. camemberti cluster. Expression of easH and easQ in the chanoclavine-I accumulating N. fumigata resulted in the accumulation of a pair of compounds of m/z 269.1 in positive mode LC-MS. Since this m/z is consistent with the mass of the isomeric pair of [rugulovasine A/B + H], we analyzed a culture of the related rugulovasine producer Penicillium biforme and found the same isomeric pair of analytes. Fragmentation of the analytes yielded fragments typical of those resulting from fragmentation of rugulovasine A/B. The deduced activities of the products of easH and easQ catalyze theoretical reactions that provide a reasonable pathway from the precursor chanoclavine-I aldehyde to the products rugulovasine A/B. When individually studied, the chanoclavine-I aldehydeaccumulating mutant of N. fumigata transformed with the P. camemberti easQ gene yielded a ratio of analytes at m/z 271.1 vs. m/z 255.1 that support the theorized mechanisms of easH and easQ to produce rugulovasines. The P. biforme genome contains a homologous eas cluster with only 12 mutations relative to that of P. camemberti. Eleven of the 12 mutations were investigated via complementation studies of the mutated genes (dmaW, easC, easE) using knockout strains of N. fumigata. The data indicate that P. camemberti has the genes to produce the ergot alkaloid rugulovasine A/B but that during domestication isolates that failed to produce alkaloids were selected for.

show abstract

“…Both 2‐ and 7‐ positions were chlorinated. However, the corresponding enzymes were not identified, and the characterization results would be promising if the genes are targeted …”

Section: Aliphatic Chlorinated Compoundsmentioning

confidence: 99%

“…However, the correspond- 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 ing enzymes were not identified, and the characterization results would be promising if the genes are targeted. [70] Chloropupukeanolides A and B, and chloropupukeanone A are three chlorinated metabolites with pupukeanane core. They were isolated from an endophytic fungus Pestalotiopsis fici, with significant antimicrobial, antitumor, and anti-HIV activities.…”

Section: Late-stage Modificationmentioning

confidence: 99%

Chlorinated Natural Products and Related Halogenases

Zeng

Zhan

2019

Israel Journal of Chemistry

View full text Add to dashboard Cite

Halogenated compounds are well‐known for their diverse bioactivities and wide applications in pharmaceutical industry, chemical engineering and other fields. Novel halometabolites isolated from nature inspire scientists to synthesize these “chance compounds”, which were, however, usually hampered by the extreme catalytic conditions and severe selectivity challenges. In the past several decades, enzymatic reactions have been brought into attention for their efficiency, selectivity, and mild reaction conditions. Synthetic biology which is in compliance with the laws of nature to a large extent shed light on the rational biosynthesis of halogenated compounds. This article summarizes several representative categories of halogenated natural products and halogenases to seek clues to the relationship between the structures and halogenase mechanisms. Some types of halogenase convert the basic building blocks to diverse early intermediates during natural product biosynthetic processes, while others are involved in the late tailoring steps to afford the final structures of halogenated molecules. The broad substrate specificity and strong regioselectivity exhibited in some halogenases are irreplaceable advantages for biosynthesis. In addition, tremendous efforts and encouraging results enable us to engineer the enzymes or techniques to synthesize “unnatural” halogenated natural products. More reaction mechanisms and structural data are expected to be revealed exponentially in this rapid growing realm.

show abstract

Dichlorinated and Brominated Rugulovasines, Ergot Alkaloids Produced by Talaromyces wortmannii

Cited by 11 publications

References 38 publications

Ergot Alkaloid Synthesis Capacity of Penicillium camemberti

Ergot Alkaloid Synthesis Capacity of Penicillium camemberti

Ergot Alkaloid Synthetic Capacity of Penicillium camemberti

Chlorinated Natural Products and Related Halogenases

Contact Info

Product

Resources

About