Psychoactive Plant Database - Neuroactive Phytochemical Collection

1. Appl Microbiol Biotechnol. 2024 May 7;108(1):323. doi: 
10.1007/s00253-024-13157-8.

An unexpected role of EasD(af): catalyzing the conversion of chanoclavine 
aldehyde to chanoclavine acid.

Yu ZP(1)(2), An C(3), Yao Y(4), Yan JZ(5), Gao SS(5), Gu YC(6)(7), Wang 
CY(8)(9)(10), Cui C(11).

Author information:
(1)Key Laboratory of Marine Drugs, The Ministry of Education of China, Institute 
of Evolution & Marine Biodiversity, School of Medicine and Pharmacy, Ocean 
University of China, Qingdao, 266003, People's Republic of China.
(2)Beijing Institute for Drug Control, NMPA Key Laboratory for Research and 
Evaluation of Generic Drugs, Beijing Key Laboratory of Analysis and Evaluation 
on Chinese Medicine, Beijing, 102206, People's Republic of China.
(3)Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 
Tianjin, 300308, People's Republic of China.
(4)State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy 
of Sciences, Beijing, 100101, People's Republic of China.
(5)Laboratory for Marine Drugs and Bioproducts, Qingdao National Laboratory for 
Marine Science and Technology, Qingdao, 266003, People's Republic of China.
(6)Laboratory for Marine Drugs and Bioproducts, Qingdao National Laboratory for 
Marine Science and Technology, Qingdao, 266003, People's Republic of China. 
yucheng.gu@syngenta.com.
(7)Syngenta Jealott's Hill International Research Centre, Bracknell, Berkshire, 
RG42 6EY, UK. yucheng.gu@syngenta.com.
(8)Key Laboratory of Marine Drugs, The Ministry of Education of China, Institute 
of Evolution & Marine Biodiversity, School of Medicine and Pharmacy, Ocean 
University of China, Qingdao, 266003, People's Republic of China. 
changyun@ouc.edu.cn.
(9)Laboratory for Marine Drugs and Bioproducts, Qingdao National Laboratory for 
Marine Science and Technology, Qingdao, 266003, People's Republic of China. 
changyun@ouc.edu.cn.
(10)Beijing Institute for Drug Control, NMPA Key Laboratory for Research and 
Evaluation of Generic Drugs, Beijing Key Laboratory of Analysis and Evaluation 
on Chinese Medicine, Beijing, 102206, People's Republic of China. 
changyun@ouc.edu.cn.
(11)Laboratory for Marine Drugs and Bioproducts, Qingdao National Laboratory for 
Marine Science and Technology, Qingdao, 266003, People's Republic of China. 
cuichs@tib.cas.cn.

Ergot alkaloids (EAs) are a diverse group of indole alkaloids known for their 
complex structures, significant pharmacological effects, and toxicity to plants. 
The biosynthesis of these compounds begins with chanoclavine-I aldehyde (CC 
aldehyde, 2), an important intermediate produced by the enzyme EasDaf or its 
counterpart FgaDH from chanoclavine-I (CC, 1). However, how CC aldehyde 2 is 
converted to chanoclavine-I acid (CC acid, 3), first isolated from Ipomoea 
violacea several decades ago, is still unclear. In this study, we provide in 
vitro biochemical evidence showing that EasDaf not only converts CC 1 to CC 
aldehyde 2 but also directly transforms CC 1 into CC acid 3 through two 
sequential oxidations. Molecular docking and site-directed mutagenesis 
experiments confirmed the crucial role of two amino acids, Y166 and S153, within 
the active site, which suggests that Y166 acts as a general base for hydride 
transfer, while S153 facilitates proton transfer, thereby increasing the acidity 
of the reaction. KEY POINTS: • EAs possess complicated skeletons and are widely 
used in several clinical diseases • EasDaf belongs to the short-chain 
dehydrogenases/reductases (SDRs) and converted CC or CC aldehyde to CC acid • 
The catalytic mechanism of EasDaf for dehydrogenation was analyzed by molecular 
docking and site mutations.

© 2024. The Author(s).

DOI: 10.1007/s00253-024-13157-8
PMCID: PMC11076337
PMID: 38713233 [Indexed for MEDLINE]

Conflict of interest statement: The authors declare no competing interests.


2. Appl Environ Microbiol. 2023 Aug 30;89(8):e0079323. doi: 10.1128/aem.00793-23.
 Epub 2023 Jul 11.

Two Satellite Gene Clusters Enhance Ergot Alkaloid Biosynthesis Capacity of 
Aspergillus leporis.

Davis KA(1), Jones AM(1), Panaccione DG(1).

Author information:
(1)Division of Plant and Soil Sciences, West Virginia University, Morgantown, 
West Virginia, USA.

Ergot alkaloids are fungal specialized metabolites that are important in 
agriculture and serve as sources of several pharmaceuticals. Aspergillus leporis 
is a soil saprotroph that possesses two ergot alkaloid biosynthetic gene 
clusters encoding lysergic acid amide production. We identified two additional, 
partial biosynthetic gene clusters within the A. leporis genome containing some 
of the ergot alkaloid synthesis (eas) genes required to make two groups of 
clavine ergot alkaloids, fumigaclavines and rugulovasines. Clavines possess 
unique biological properties compared to lysergic acid derivatives. 
Bioinformatic analyses indicated the fumigaclavine cluster contained functional 
copies of easA, easG, easD, easM, and easN. Genes resembling easQ and easH, 
which are required for rugulovasine production, were identified in a separate 
gene cluster. The pathways encoded by these partial, or satellite, clusters 
would require intermediates from the previously described lysergic acid amide 
pathway to synthesize a product. Chemical analyses of A. leporis cultures 
revealed the presence of fumigaclavine A. However, rugulovasine was only 
detected in a single sample, prompting a heterologous expression approach to 
confirm functionality of easQ and easH. An easA knockout strain of Metarhizium 
brunneum, which accumulates the rugulovasine precursor chanoclavine-I aldehyde, 
was chosen as expression host. Strains of M. brunneum expressing easQ and easH 
from A. leporis accumulated rugulovasine as demonstrated through mass 
spectrometry analysis. These data indicate that A. leporis is exceptional among 
fungi in having the capacity to synthesize products from three branches of the 
ergot alkaloid pathway and for utilizing an unusual satellite cluster approach 
to achieve that outcome. IMPORTANCE Ergot alkaloids are chemicals produced by 
several species of fungi and are notable for their impacts on agriculture and 
medicine. The ability to make ergot alkaloids is typically encoded by a 
clustered set of genes that are physically adjacent on a chromosome. Different 
ergot alkaloid classes are formed via branching of a complex pathway that begins 
with a core set of the same five genes. Most ergot alkaloid-producing fungi have 
a single cluster of genes that is complete, or self-sufficient, and produce 
ergot alkaloids from one or occasionally two branches from that single cluster. 
Our data show that Aspergillus leporis is exceptional in having the genetic 
capacity to make products from three pathway branches. Moreover, it uses a 
satellite cluster approach, in which gene products of partial clusters rely on 
supplementation with a chemical intermediate produced via another gene cluster, 
to diversify its biosynthetic potential without duplicating all the steps.

DOI: 10.1128/aem.00793-23
PMCID: PMC10467348
PMID: 37432119 [Indexed for MEDLINE]

Conflict of interest statement: The authors declare no conflict of interest.


3. ACS Synth Biol. 2023 Apr 21;12(4):1133-1145. doi: 10.1021/acssynbio.2c00626. 
Epub 2023 Mar 29.

Modular Pathway Compartmentalization for Agroclavine Overproduction in 
Saccharomyces cerevisiae.

Wu N(1)(2), Yao M(1)(2), Xiao W(1)(2), Dong T(1)(2), Ma H(1)(2), Du X(3), Wang 
Y(1)(2), Yuan Y(1)(2).

Author information:
(1)Frontier Science Center for Synthetic Biology and Key Laboratory of Systems 
Bioengineering (Ministry of Education), School of Chemical Engineering and 
Technology, Tianjin University, Tianjin 300072, China.
(2)Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin 
University, Tianjin 300072, China.
(3)State Key Laboratory of Toxicology and Medical Countermeasures, Beijing 
Institute of Pharmacology and Toxicology, 100850 Beijing, China.

Agroclavine, which has anti-depressant activity and anti-Alzheimer effects, is 
the raw material used to synthesize ergo-based drugs. Although the production of 
agroclavine from Saccharomyces cerevisiae is possible, its yield is 
exceptionally low. The current study proposes a modular compartmentalization 
strategy for identifying and modifying the bottleneck step in agroclavine 
overproduction. The agroclavine synthetic pathway was reconstituted in yeast, 
and the best combination of Claviceps fusiformis EasA with Claviceps purpurea 
EasD/EasG was identified. According to the data on the expression and 
subcellular localization of agroclavine pathway proteins, the whole pathway was 
divided into two modules by chanoclavine-I. Separate enzyme distribution within 
the downstream module and low expression of DmaW and EasE in the upstream module 
were identified as the bottleneck steps in the pathway. The pathway efficiency 
was enhanced 2.06-fold when the downstream module was entirely anchored to the 
endoplasmic reticulum compartment. Increasing NADPH supply by overexpressing 
POS5 further improved the agroclavine yield by 27.4%. Altering the intracellular 
localization of DmaW from the peroxisome to the endoplasmic reticulum (ER) not 
only improved protein expression but also accelerated the accumulation of 
agroclavine by 59.9%. Integration of all modified modules into the host 
chromosome resulted in an improved yield of agroclavine at 101.6 mg/L with flask 
fermentation (a 241-fold improvement over the initial strain) and ultimately 
produced 152.8 mg/L of agroclavine on fed-batch fermentation. The current study 
unlocked the potential of S. cerevisiae in the advanced biosynthesis of ergot 
alkaloids. It also provides a promising strategy to reconstitute 
compartmentalized pathways.

DOI: 10.1021/acssynbio.2c00626
PMID: 36987837 [Indexed for MEDLINE]


4. Mycologia. 2020 Sep-Oct;112(5):974-988. doi: 10.1080/00275514.2020.1797372.
Epub  2020 Sep 16.

Four phylogenetic species of ergot from Canada and their characteristics in 
morphology, alkaloid production, and pathogenicity.

Liu M(1), Overy DP(1), Cayouette J(1), Shoukouhi P(1), Hicks C(1), Bisson K(1), 
Sproule A(1), Wyka SA(2), Broders K(2), Popovic Z(3), Menzies JG(3).

Author information:
(1)Ottawa Research and Development Centre, Agriculture and Agri-Food Canada , 
Ottawa, K1A 0C6, Canada.
(2)Colorado State University , Fort Collins, Colorado 80523.
(3)Morden Research and Development Centre, Agriculture and Agri-Food Canada , 
101 Route 100, Morden, Manitoba R6M 1Y5, Canada.

Four ergot species (Claviceps ripicola, C. quebecensis, C. perihumidiphila, and 
C. occidentalis) were recognized based on analyses of DNA sequences from 
multiple loci, including two housekeeping genes, RNA polymerase II second 
largest subunit (RPB2), and translation elongation factor 1-α (TEF1-α), and a 
single-copy ergot alkaloid synthesis gene (easE) encoding chanoclavine I 
synthase oxidoreductase. Morphological features, ergot alkaloid production, and 
pathogenicity on five common cereal crops of each species were evaluated and 
presented in taxonomic descriptions. A synoptic key was also provided for 
identification.

DOI: 10.1080/00275514.2020.1797372
PMID: 32936061 [Indexed for MEDLINE]


5. Appl Environ Microbiol. 2018 Sep 17;84(19):e01583-18. doi:
10.1128/AEM.01583-18.  Print 2018 Oct 1.

Ergot Alkaloid Synthesis Capacity of Penicillium camemberti.

Fabian SJ(1), Maust MD(1), Panaccione DG(2).

Author information:
(1)West Virginia University, Division of Plant and Soil Sciences, Genetics and 
Developmental Biology Program, Morgantown, West Virginia, USA.
(2)West Virginia University, Division of Plant and Soil Sciences, Genetics and 
Developmental Biology Program, Morgantown, West Virginia, USA danpan@wvu.edu.

Ergot alkaloids are specialized fungal metabolites with potent biological 
activities. They are encoded by well-characterized gene clusters in the genomes 
of producing fungi. Penicillium camemberti plays a major role in the ripening of 
Brie and Camembert cheeses. The P. camemberti 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 (easH and 
easQ) 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 P. camemberti, and did not detect chanoclavine-I aldehyde or its 
derivatives. To create a functioning facsimile of the P. camembertieas cluster, 
we expressed P. camemberti easH and easQ in a chanoclavine-I 
aldehyde-accumulating easA knockout mutant of Neosartorya fumigata The 
easH-easQ-engineered N. fumigata strain accumulated a pair of compounds of m/z 
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 Penicillium biforme 
accumulated the same isomeric pair of analytes. The P. camemberti eas genes were 
transcribed in culture, but comparison of the P. camemberti eas cluster with the 
functional cluster from P. biforme indicated 11 polymorphisms. Whereas other P. 
camembertieas genes functioned when expressed in N. fumigata, P. camembertieasC 
did not restore ergot alkaloids when expressed in an easC mutant. The data 
indicate that P. camemberti formerly had the capacity to produce the ergot 
alkaloids rugulovasine A and B.IMPORTANCE The presence of ergot alkaloid 
synthesis genes in the genome of Penicillium camemberti 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. Penicillium biforme, a close relative of P. 
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 the P. camemberti 
pathway in the model fungus Neosartorya fumigata indicated that P. camemberti 
formerly had the capacity to produce these same ergot alkaloids. Neither P. 
camemberti nor P. biforme produced ergot alkaloids in cheese, indicating that 
nutritionally driven gene regulation prevents these fungi from producing ergot 
alkaloids in a dairy environment.

Copyright © 2018 American Society for Microbiology.

DOI: 10.1128/AEM.01583-18
PMCID: PMC6146994
PMID: 30076193 [Indexed for MEDLINE]