scholarly journals An Ergot Alkaloid Biosynthesis Gene and Clustered Hypothetical Genes from Aspergillus fumigatus

2005 ◽  
Vol 71 (6) ◽  
pp. 3112-3118 ◽  
Author(s):  
Christine M. Coyle ◽  
Daniel G. Panaccione
Keyword(s):  
Aspergillus Fumigatus ◽  
Ergot Alkaloid ◽  
Biological Activities ◽  
Ergot Alkaloids ◽  
Alkaloid Production ◽  
Genetic Origin ◽  
Biosynthesis Gene ◽  
Opportunistic Human Pathogen ◽  
Clavicipitaceous Fungi ◽  
Hypothetical Genes

ABSTRACT The ergot alkaloids are a family of indole-derived mycotoxins with a variety of significant biological activities. Aspergillus fumigatus, a common airborne fungus and opportunistic human pathogen, and several fungi in the relatively distant taxon Clavicipitaceae (clavicipitaceous fungi) produce different sets of ergot alkaloids. The ergot alkaloids of these divergent fungi share a four-member ergoline ring but differ in the number, type, and position of the side chains. Several genes required for ergot alkaloid production are known in the clavicipitaceous fungi, and these genes are clustered in the genome of the ergot fungus Claviceps purpurea. We investigated whether the ergot alkaloids of A. fumigatus have a common biosynthetic and genetic origin with those of the clavicipitaceous fungi. A homolog of dmaW, the gene controlling the determinant step in the ergot alkaloid pathway of clavicipitaceous fungi, was identified in the A. fumigatus genome. Knockout of dmaW eliminated all known ergot alkaloids from A. fumigatus, and complementation of the mutation restored ergot alkaloid production. Clustered with dmaW in the A. fumigatus genome are sequences corresponding to five genes previously proposed to encode steps in the ergot alkaloid pathway of C. purpurea, as well as additional sequences whose deduced protein products are consistent with their involvement in the ergot alkaloid pathway. The corresponding genes have similarities in their nucleotide sequences, but the orientations and positions within the cluster of several of these genes differ. The data indicate that the ergot alkaloid biosynthetic capabilities in A. fumigatus and the clavicipitaceous fungi had a common origin.

scholarly journals An Old Yellow Enzyme Gene Controls the Branch Point between Aspergillus fumigatus and Claviceps purpurea Ergot Alkaloid Pathways

2010 ◽  
Vol 76 (12) ◽  
pp. 3898-3903 ◽  
Author(s):  
Christine M. Coyle ◽  
Johnathan Z. Cheng ◽  
Sarah E. O'Connor ◽  
Daniel G. Panaccione
Keyword(s):  
Aspergillus Fumigatus ◽  
Ergot Alkaloid ◽  
Ergot Alkaloids ◽  
Aldehyde Group ◽  
Ring Structure ◽  
Ring Closure ◽  
Claviceps Purpurea ◽  
Wild Type ◽  
Old Yellow Enzyme ◽  
Clavicipitaceous Fungi

ABSTRACT Ergot fungi in the genus Claviceps and several related fungal groups in the family Clavicipitaceae produce toxic ergot alkaloids. These fungi produce a variety of ergot alkaloids, including clavines as well as lysergic acid derivatives. Ergot alkaloids are also produced by the distantly related, opportunistic human pathogen Aspergillus fumigatus. However, this fungus produces festuclavine and fumigaclavines A, B, and C, which collectively differ from clavines of clavicipitaceous fungi in saturation of the last assembled of four rings in the ergoline ring structure. The two lineages are hypothesized to share early steps of the ergot alkaloid pathway before diverging at some point after the synthesis of the tricyclic intermediate chanoclavine-I. Disruption of easA, a gene predicted to encode a flavin-dependent oxidoreductase of the old yellow enzyme class, in A. fumigatus led to accumulation of chanoclavine-I and chanoclavine-I-aldehyde. Complementation of the A. fumigatus easA mutant with a wild-type allele from the same fungus restored the wild-type profile of ergot alkaloids. These data demonstrate that the product of A. fumigatus easA is required for incorporation of chanoclavine-I-aldehyde into more-complex ergot alkaloids, presumably by reducing the double bond conjugated to the aldehyde group, thus facilitating ring closure. Augmentation of the A. fumigatus easA mutant with a homologue of easA from Claviceps purpurea resulted in accumulation of ergot alkaloids typical of clavicipitaceous fungi (agroclavine, setoclavine, and its diastereoisomer isosetoclavine). These data indicate that functional differences in the easA-encoded old yellow enzymes of A. fumigatus and C. purpurea result in divergence of their respective ergot alkaloid pathways.

SOME ASPECTS OF PHOSPHATE METABOLISM OF CLAVICEPS PURPUREA (FR.) TUL.: II. RELATIONSHIP BETWEEN NUCLEIC ACIDS SYNTHESIS AND ERGOT ALKALOID PRODUCTION

1961 ◽  
Vol 7 (6) ◽  
pp. 883-888 ◽  
Author(s):  
C. de Waart
Keyword(s):  
Nucleic Acid ◽  
Ergot Alkaloid ◽  
Ribonucleic Acid ◽  
Ergot Alkaloids ◽  
Acid Synthesis ◽  
Claviceps Purpurea ◽  
Phosphate Metabolism ◽  
Alkaloid Production ◽  
Phosphorus Containing ◽  
Insoluble Phosphate

A study was made of the distribution of the phosphorus-containing compounds in the phosphate pool of stationary cultures of Claviceps purpurea (Fr.) Tul. Exogenous KH2PO4 was mainly converted to ribonucleic acid. Conditions favorable to increased nucleic acid synthesis increased the yield of ergot alkaloids. The proportion of nucleic acid and acid-insoluble phosphate fraction appeared to be an important factor influencing the ergot alkaloid production.

scholarly journals Ergot Alkaloid Synthesis Capacity ofPenicillium camemberti

2018 ◽  
Vol 84 (19) ◽  
Author(s):  
Samantha J. Fabian ◽  
Matthew D. Maust ◽  
Daniel G. Panaccione
Keyword(s):  
Ergot Alkaloid ◽  
Biological Activities ◽  
Ergot Alkaloids ◽  
Gene Clusters ◽  
Close Relative ◽  
Knockout Mutant ◽  
Content Type ◽  
Penicillium Camemberti ◽  
Alkaloid Synthesis ◽  
Positive Mode

ABSTRACTErgot alkaloids are specialized fungal metabolites with potent biological activities. They are encoded by well-characterized gene clusters in the genomes of producing fungi.Penicillium camembertiplays a major role in the ripening of Brie and Camembert cheeses. TheP. camembertigenome 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 (easHandeasQ) that may control modification of chanoclavine-I aldehyde into other ergot alkaloids. We analyzed samples of Brie and Camembert cheeses, as well as cultures ofP. camemberti, and did not detect chanoclavine-I aldehyde or its derivatives. To create a functioning facsimile of theP. camembertieascluster, we expressedP. camemberti easHandeasQin a chanoclavine-I aldehyde-accumulatingeasAknockout mutant ofNeosartorya fumigata. TheeasH-easQ-engineeredN. fumigatastrain accumulated a pair of compounds ofm/z269.1288 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 producerPenicillium biformeaccumulated the same isomeric pair of analytes. TheP. camemberti easgenes were transcribed in culture, but comparison of theP. camemberti eascluster with the functional cluster fromP. biformeindicated 11 polymorphisms. Whereas otherP. camembertieasgenes functioned when expressed inN. fumigata,P. camembertieasCdid not restore ergot alkaloids when expressed in aneasCmutant. The data indicate thatP. camembertiformerly had the capacity to produce the ergot alkaloids rugulovasine A and B.IMPORTANCEThe presence of ergot alkaloid synthesis genes in the genome ofPenicillium camembertiis 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.Penicillium biforme, a close relative ofP. camemberti, 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 of theP. camembertipathway in the model fungusNeosartorya fumigataindicated thatP. camembertiformerly had the capacity to produce these same ergot alkaloids. NeitherP. camembertinorP. biformeproduced ergot alkaloids in cheese, indicating that nutritionally driven gene regulation prevents these fungi from producing ergot alkaloids in a dairy environment.

Ergot alkaloid biosynthesis by isolates of Balansia epichloë and B. henningsiana

1981 ◽  
Vol 59 (12) ◽  
pp. 2534-2538 ◽  
Author(s):  
Charles W. Bacon ◽  
James K. Porter ◽  
Joe D. Robbins
Keyword(s):  
Ergot Alkaloid ◽  
Sampling Site ◽  
Total Yield ◽  
Ergot Alkaloids ◽  
Chemical Species ◽  
Alkaloid Biosynthesis ◽  
Andropogon Virginicus ◽  
Clavicipitaceous Fungi

Two endophytic clavicipitaceous fungi, Balansia epichloë and B. henningsiana, were isolated from several grasses, cultured on laboratory media, and assessed for their ability to produce the major ergot alkaloids agroclavine, chanoclavine (I), ergonovine, and ergonovinine. Data indicated that the ability of these two species of fungi to biosynthesize ergot alkaloids was host related. All isolates of B. epichloë from smut-grass (Sporobolus poiretii) and 59% of the isolates of B. henningsiana from broom-sedge (Andropogon virginicus) produced the major alkaloids in culture. All isolates of B. epichloë from Eragrostis secundiflora and E. hirsuta, and all isolates of B. henningsiana from Panicum tenerum failed to produce alkaloids. The total yield and chemical species of individual alkaloids varied among isolates from a sampling site.

scholarly journals Abundant Respirable Ergot Alkaloids from the Common Airborne Fungus Aspergillus fumigatus

2005 ◽  
Vol 71 (6) ◽  
pp. 3106-3111 ◽  
Author(s):  
Daniel G. Panaccione ◽  
Christine M. Coyle
Keyword(s):  
Physical Properties ◽  
Aspergillus Fumigatus ◽  
Ergot Alkaloids ◽  
Broth Culture ◽  
Latex Paint ◽  
Adverse Health Effects ◽  
Airborne Fungus ◽  
Monoamine Receptors ◽  
High Concentrations ◽  
Opportunistic Human Pathogen

ABSTRACT Ergot alkaloids are mycotoxins that interact with several monoamine receptors, negatively affecting cardiovascular, nervous, reproductive, and immune systems of exposed humans and animals. Aspergillus fumigatus, a common airborne fungus and opportunistic human pathogen, can produce ergot alkaloids in broth culture. The objectives of this study were to determine if A. fumigatus accumulates ergot alkaloids in a respirable form in or on its conidia, to quantify ergot alkaloids associated with conidia produced on several different substrates, and to measure relevant physical properties of the conidia. We found at least four ergot alkaloids, fumigaclavine C, festuclavine, fumigaclavine A, and fumigaclavine B (in order of abundance), associated with conidia of A. fumigatus. Under environmentally relevant conditions, the total mass of ergot alkaloids often constituted >1% of the mass of the conidium. Ergot alkaloids were extracted from conidia produced on all media tested, and the greatest quantities were observed when the fungus was cultured on latex paint or cultured maize seedlings. The values for physical properties of conidia likely to affect their respirability (i.e., diameter, mass, and specific gravity) were significantly lower for A. fumigatus than for Aspergillus nidulans, Aspergillus niger, and Stachybotrys chartarum. The demonstration of relatively high concentrations of ergot alkaloids associated with conidia of A. fumigatus presents opportunities for investigations of potential contributions of the toxins to adverse health effects associated with the fungus and to aspects of the biology of the fungus that contribute to its success.

scholarly journals A Baeyer-Villiger monooxygenase gene involved in the synthesis of lysergic acid amides affects the interaction of the fungus Metarhizium brunneum with insects

2021 ◽  
Author(s):  
Chey R. Steen ◽  
Jessi K. Sampson ◽  
Daniel G. Panaccione
Keyword(s):  
Ergot Alkaloid ◽  
Galleria Mellonella ◽  
Insect Pests ◽  
Biological Activities ◽  
Plant Root ◽  
Total Concentration ◽  
Acid Amide ◽  
Ergot Alkaloids ◽  
Structural Similarity ◽  
Lysergic Acid

Several fungi, including the plant root symbiont and insect pathogen Metarhizium brunneum , produce lysergic acid amides via a branch of the ergot alkaloid pathway. Lysergic acid amides include important pharmaceuticals and pharmaceutical lead compounds and have potential ecological significance, making knowledge of their biosynthesis relevant. Many steps in the biosynthesis of lysergic acid amides have been determined, but terminal steps in the synthesis of lysergic acid α-hydroxyethylamide (LAH)––by far the most abundant lysergic acid amide in M. brunneum ––are unknown. Ergot alkaloid synthesis ( eas ) genes are clustered in the genomes of fungi that produce these compounds, and the eas clusters of LAH producers contain two uncharacterized genes ( easO and easP ) not found in fungi that do not produce LAH. Knockout of easO via a CRISPR-Cas9 approach eliminated LAH and resulted in accumulation of alternate lysergic acid amides lysergyl-alanine and ergonovine. Despite the elimination of LAH, the total concentration of lysergic acid derivatives was not affected significantly by the mutation. Complementation with a wild-type allele of easO restored the ability to synthesize LAH. Substrate feeding studies indicated that neither lysergyl-alanine nor ergonovine were substrates for the product of easO (EasO). EasO had structural similarity to Baeyer-Villiger monooxygenases (BVMOs), and labeling studies with deuterated alanine supported a role for a BVMO in LAH biosynthesis. The easO knockout had reduced virulence to larvae of the insect Galleria mellonella , indicating that LAH contributes to virulence of M. brunneum on insects and that LAH has biological activities different from ergonovine and lysergyl-alanine. Importance Fungi in the genus Metarhizium are important plant root symbionts and insect pathogens. They are formulated commercially to protect plants from insect pests. Several Metarhizium species including M. brunneum were recently shown to produce ergot alkaloids, a class of specialized metabolites studied extensively in other fungi because of their importance in agriculture and medicine. A biological role for ergot alkaloids in Metarhizium species had not been demonstrated previously. Moreover, the types of ergot alkaloids produced by Metarhizium species are lysergic acid amides, which have served directly or indirectly as important pharmaceutical compounds. The terminal steps in the synthesis of the most abundant lysergic acid amide in Metarhizium species and several other fungi (LAH) have not been determined. The results of this study demonstrate the role of a previously unstudied gene in LAH synthesis and indicate that LAH contributes to virulence of M. brunneum on insects.

scholarly journals Links Between Genetic Groups, Host Specificity, and Ergot-Alkaloid Profiles within Claviceps purpurea (Fr.) Tul. on Slovenian Grasses

Plant Disease ◽  
2018 ◽  
Vol 102 (7) ◽  
pp. 1334-1340
Author(s):  
Matevž Likar ◽  
Marjana Grandič ◽  
Breda Jakovac Strajn ◽  
Katarina Kos ◽  
Franci Aco Celar
Keyword(s):  
Host Specificity ◽  
Ribosomal Dna ◽  
Ergot Alkaloid ◽  
Animal Feed ◽  
Genetic Relationships ◽  
Ergot Alkaloids ◽  
Claviceps Purpurea ◽  
Alkaloid Production ◽  
Alkaloid Composition ◽  
Genetic Groups

In the present study, the genetic relationships and ergot-alkaloid production of the fungus Claviceps purpurea on grasses were investigated, to determine any associations between grass host specificity, ergot-alkaloid production, and geographic origin. C. purpurea sclerotia were obtained from wild and cultivated grasses along a 300-km climatic gradient, from sub-Mediterranean to continental climates. Twenty-one infected grass samples provided 39 sclerotia for analysis of the ergot alkaloids ergometrine, ergosine, ergotamine, ergocornine, ergocryptine, and ergocristine, and their “-inine” epimers, using liquid chromatography–tandem mass spectrometry. C. purpurea ribosomal DNA underwent molecular classification to determine any grass host or geographic specificity of ergot-alkaloid composition for the different operational taxonomic units. Molecular analysis of sclerotia ribosomal DNA showed three genetic groups, with some associations with specific grass host taxonomic groups. The ergot-alkaloid composition data were in agreement with the data obtained by molecular methods. The most frequent ergot-alkaloid epimers were ergocristine, and ergosine. The total ergot-alkaloid concentrations in sclerotia varied from 59 to 4,200 mg kg–1, which corresponds to 0.059 to 4.2 mg kg–1 in animal feed (assuming ergot alkaloids at 1,000 mg kg–1 sclerotia). Therefore, grasses can be associated with significant levels of ergot alkaloids. In addition, the ergot-alkaloid compositions of C. purpurea sclerotia can be different for infections with different C. purpurea genetic groups, because these show different ergot-alkaloid compositions.

scholarly journals Assessment of ergot (Claviceps purpurea) exposure in pregnant and postpartum beef cows

2018 ◽  
Vol 98 (4) ◽  
pp. 688-700 ◽  
Author(s):  
T. Grusie ◽  
V. Cowan ◽  
J. Singh ◽  
J. McKinnon ◽  
B. Blakley
Keyword(s):  
Ergot Alkaloid ◽  
Post Partum ◽  
Ergot Alkaloids ◽  
Maximum Size ◽  
Beef Cows ◽  
Plasma Prolactin ◽  
Claviceps Purpurea ◽  
Ambient Temperatures ◽  
Beef Cow ◽  
The Impact

Cows were fed ration for 9 wk containing 5, 48, 201, and 822 μg kg−1 ergot alkaloids. The objective was to evaluate the impact of ergot consumption in beef cow–calf operations. Ergot alkaloids up to 822 μg kg−1 did not alter the weight of peripartum and postpartum beef cows (P = 0.93) or nursing calves (P = 0.08), rectal temperature (P = 0.16), or plasma prolactin concentrations (P = 0.30) at moderate ambient temperatures. Ergot did not influence the time (>1 ng mL−1; P = 0.79) or the progesterone concentration (P = 0.38) at the time of first postpartum rise or the size of the first (14 ± 0.6 mm; P = 0.40) and second (13 ± 0.5 mm; P = 0.41) follicles to ovulate. The maximum size of the first postpartum corpus luteum (CL) was 4 mm larger in the 822 μg kg−1 ergot group compared with the control (P = 0.03) for the first ovulation post partum, but not for the second (P = 0.11). There was no effect of ergot exposure on the number of days until the appearance of the first (43 ± 4 d; P = 0.95) or second (52 ± 4 d; P = 0.98) CL post partum. Ergot alkaloid concentrations up to 822 μg kg−1 did not affect pregnancy rates (X2 = 0.36). In conclusion, ergot alkaloid exposure for 9 wk to concentrations as high as 822 μg kg−1 did not alter performance in pregnant and postpartum beef cattle at moderate ambient temperatures.

scholarly journals Heterologous Expression of Lysergic Acid and Novel Ergot Alkaloids in Aspergillus fumigatus

2014 ◽  
Vol 80 (20) ◽  
pp. 6465-6472 ◽  
Author(s):  
Sarah L. Robinson ◽  
Daniel G. Panaccione
Keyword(s):  
Aspergillus Fumigatus ◽  
Candidate Genes ◽  
Mass Spectra ◽  
Ergot Alkaloids ◽  
Lysergic Acid ◽  
Precursor Feeding ◽  
Gene Encoding ◽  
Content Type ◽  
Multiple Reactions ◽  
Important Pathway

ABSTRACTDifferent lineages of fungi produce distinct classes of ergot alkaloids. Lysergic acid-derived ergot alkaloids produced by fungi in the Clavicipitaceae are particularly important in agriculture and medicine. The pathway to lysergic acid is partly elucidated, but the gene encoding the enzyme that oxidizes the intermediate agroclavine is unknown. We investigated two candidate agroclavine oxidase genes from the fungusEpichloë festucaevar.lolii×Epichloë typhinaisolate Lp1 (henceforth referred to asEpichloësp. Lp1), which produces lysergic acid-derived ergot alkaloids. Candidate geneseasHandcloAwere expressed in a mutant strain of the moldAspergillus fumigatus, which typically produces a subclass of ergot alkaloids not derived from agroclavine or lysergic acid. Candidate genes were coexpressed with theEpichloësp. Lp1 allele ofeasA, which encodes an enzyme that catalyzed the synthesis of agroclavine from anA. fumigatusintermediate; the agroclavine then served as the substrate for the candidate agroclavine oxidases. Strains expressingeasAandcloAfromEpichloësp. Lp1 produced lysergic acid from agroclavine, a process requiring a cumulative six-electron oxidation and a double-bond isomerization. Strains that accumulated excess agroclavine (as a result ofEpichloësp. Lp1easAexpression in the absence ofcloA) metabolized it into two novel ergot alkaloids for which provisional structures were proposed on the basis of mass spectra and precursor feeding studies. Our data indicate that CloA catalyzes multiple reactions to produce lysergic acid from agroclavine and that combining genes from different ergot alkaloid pathways provides an effective strategy to engineer important pathway molecules and novel ergot alkaloids.

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