Psychoactive Plant Database - Neuroactive Phytochemical Collection

1. Front Microbiol. 2023 Mar 16;14:1150937. doi: 10.3389/fmicb.2023.1150937. 
eCollection 2023.

Increasing lysergic acid levels for ergot alkaloid biosynthesis: Directing 
catalysis via the F-G loop of Clavine oxidases.

Lim LR(1)(2)(3)(4), Wong G(1)(2)(4), Go MK(1)(2)(4), Yew WS(1)(2)(3)(4).

Author information:
(1)Synthetic Biology for Clinical and Technological Innovation, National 
University of Singapore, Singapore, Singapore.
(2)Synthetic Biology Translational Research Programme, Yong Loo Lin School of 
Medicine, National University of Singapore, Singapore, Singapore.
(3)NUS Graduate School-Integrative Sciences and Engineering Programme (ISEP), 
National University of Singapore, Singapore, Singapore.
(4)Department of Biochemistry, Yong Loo Lin School of Medicine, National 
University of Singapore, Singapore, Singapore.

Most ergot alkaloid drugs are semi-synthetically derived from the natural 
product lysergic acid, a valuable precursor for the development of novel ergot 
alkaloid drugs. Clavine oxidase (CloA) is a putative cytochrome P450, identified 
in the ergot alkaloid biosynthesis pathway, and a key enzyme that catalyzes the 
formation of lysergic acid from the precursor alkaloid agroclavine in a two-step 
oxidation reaction. We demonstrated in this study that Saccharomyces cerevisiae 
can be used as a viable host for the functional expression of CloA from 
Claviceps purpurea and its orthologs. We also showed that CloA orthologs differ 
in their ability to oxidize the substrate agroclavine, with some orthologs only 
able to perform the first oxidation reaction to produce elymoclavine. Of 
particular note, we identified a region between the F-G helices of the enzyme 
that may be involved in directing oxidation of agroclavine by substrate 
recognition and uptake. Using this knowledge, engineered CloAs were shown to 
produce lysergic acid at levels exceeding that of wildtype CloA orthologs; a 
CloA variant, chimeric AT5 9Hypo CloA, increased production levels of lysergic 
acid to 15 times higher as compared to the wildtype enzyme, demonstrating future 
utility for the industrial production of ergot alkaloids using biosynthetic 
routes.

Copyright © 2023 Lim, Wong, Go and Yew.

DOI: 10.3389/fmicb.2023.1150937
PMCID: PMC10060963
PMID: 37007471

Conflict of interest statement: The authors declare that the research was 
conducted in the absence of any commercial or financial relationships that could 
be construed as a potential conflict of interest.


2. Appl Microbiol Biotechnol. 2022 Apr;106(8):2981-2991. doi: 
10.1007/s00253-022-11892-4. Epub 2022 Apr 7.

Beyond the cyclopropyl ring formation: fungal Aj_EasH catalyzes asymmetric 
hydroxylation of ergot alkaloids.

An C(#)(1)(2), Zhu F(#)(3)(4), Yao Y(#)(4), Zhang K(4)(5), Wang W(4)(5), Zhang 
J(4), Wei G(4)(5), Xia Y(4), Gao Q(6), Gao SS(7).

Author information:
(1)Key Laboratory of Industrial Fermentation Microbiology of Ministry of 
Education, Biotechnology College of Tianjin University of Science and 
Technology, Tianjin, 300457, China. ancy@im.ac.cn.
(2)State Key Laboratory of Microbial Resources, Institute of Microbiology, 
Chinese Academy of Sciences, Beijing, 100101, People's Republic of China. 
ancy@im.ac.cn.
(3)Key Laboratory of Industrial Fermentation Microbiology of Ministry of 
Education, Biotechnology College of Tianjin University of Science and 
Technology, Tianjin, 300457, China.
(4)State Key Laboratory of Microbial Resources, Institute of Microbiology, 
Chinese Academy of Sciences, Beijing, 100101, People's Republic of China.
(5)University of Chinese Academy of Sciences, Beijing, 100049, People's Republic 
of China.
(6)Key Laboratory of Industrial Fermentation Microbiology of Ministry of 
Education, Biotechnology College of Tianjin University of Science and 
Technology, Tianjin, 300457, China. gaoqiang@tust.edu.cn.
(7)State Key Laboratory of Microbial Resources, Institute of Microbiology, 
Chinese Academy of Sciences, Beijing, 100101, People's Republic of China. 
gaoss@im.ac.cn.
(#)Contributed equally

Ergot alkaloids (EAs) are among the most important bioactive natural products. 
FeII/α-ketoglutarate-dependent dioxygenase Aj_EasH from Aspergillus japonicus is 
responsible for the formation of the cyclopropyl ring of the ergot alkaloid (EA) 
cycloclavine (4). Herein we reconstituted the biosynthesis of 4 in vitro from 
prechanoclavine (1) for the first time. Additionally, an unexpected activity of 
asymmetric hydroxylation at the C-4 position of EA compound festuclavine (5) for 
Aj_EasH was revealed. Furthermore, Aj_EasH also catalyzes the hydroxylation of 
two more EAs 9,10-dihydrolysergol (6) and elymoclavine (7). Thus, our results 
proved that Aj_EasH is a promiscuous and bimodal dioxygenase that catalyzes both 
the formation of cyclopropyl ring in 4 and the asymmetric hydroxylation of EAs. 
Molecular docking (MD) revealed the substrate-binding mode as well as the 
catalytic mechanism of asymmetric hydroxylation, suggesting more EAs could 
potentially be recognized and hydroxylated by Aj_EasH. Overall, the newly 
discovered activity empowered Aj_EasH with great potential for producing more 
diverse and bioactive EA derivatives. KEY POINTS: • Aj_EasH was revealed to be a 
promiscuous and bimodal FeII/α-ketoglutarate-dependent dioxygenase. • Aj_EasH 
converted festuclavine, 9,10-dihydrolysergol, and elymoclavine to their 
hydroxylated derivatives. • The catalytic mechanism of Aj_EasH for hydroxylation 
was analyzed by molecular docking.

© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, 
part of Springer Nature.

DOI: 10.1007/s00253-022-11892-4
PMID: 35389067 [Indexed for MEDLINE]


3. Metab Eng. 2022 Jan;69:198-208. doi: 10.1016/j.ymben.2021.12.002. Epub 2021
Dec  10.

Overproduction of medicinal ergot alkaloids based on a fungal platform.

Yao Y(1), Wang W(2), Shi W(3), Yan R(1), Zhang J(1), Wei G(2), Liu L(4), Che 
Y(5), An C(6), Gao SS(7).

Author information:
(1)State Key Laboratory of Microbial Resources, Institute of Microbiology, 
Chinese Academy of Sciences, Beijing, 100101, PR China.
(2)State Key Laboratory of Microbial Resources, Institute of Microbiology, 
Chinese Academy of Sciences, Beijing, 100101, PR China; University of Chinese 
Academy of Sciences, Beijing, 100049, PR China.
(3)Microbial Resource and Big Data Center, Institute of Microbiology, Chinese 
Academy of Sciences, Beijing, 100101, PR China.
(4)State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy 
of Sciences, Beijing, 100101, PR China.
(5)Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and 
Peking Union Medical College, Beijing, 100050, PR China.
(6)State Key Laboratory of Microbial Resources, Institute of Microbiology, 
Chinese Academy of Sciences, Beijing, 100101, PR China. Electronic address: 
ancy@im.ac.cn.
(7)State Key Laboratory of Microbial Resources, Institute of Microbiology, 
Chinese Academy of Sciences, Beijing, 100101, PR China. Electronic address: 
gaoss@im.ac.cn.

Privileged ergot alkaloids (EAs) produced by the fungal genus Claviceps are used 
to treat a wide range of diseases. However, their use and research have been 
hampered by the challenging genetic engineering of Claviceps. Here we 
systematically refactored and rationally engineered the EA biosynthetic pathway 
in heterologous host Aspergillus nidulans by using a Fungal-Yeast-Shuttle-Vector 
protocol. The obtained strains allowed the production of diverse EAs and related 
intermediates, including prechanoclavine (PCC, 333.8 mg/L), chanoclavine (CC, 
241.0 mg/L), agroclavine (AC, 78.7 mg/L), and festuclavine (FC, 99.2 mg/L), etc. 
This fungal platform also enabled the access to the methyl-oxidized EAs (MOEAs), 
including elymoclavine (EC), lysergic acid (LA), dihydroelysergol (DHLG), and 
dihydrolysergic acid (DHLA), by overexpressing a P450 enzyme CloA. Furthermore, 
by optimizing the P450 electron transfer (ET) pathway and using multi-copy of 
cloA, the titers of EC and DHLG have been improved by 17.3- and 9.4-fold, 
respectively. Beyond our demonstration of A. nidulans as a robust platform for 
EA overproduction, our study offers a proof of concept for engineering the 
eukaryotic P450s-contained biosynthetic pathways in a filamentous fungal host.

Copyright © 2021. Published by Elsevier Inc.

DOI: 10.1016/j.ymben.2021.12.002
PMID: 34902590 [Indexed for MEDLINE]


4. Food Chem Toxicol. 2018 Jun;116(Pt B):298-306. doi: 10.1016/j.fct.2018.04.024.
 Epub 2018 Apr 13.

In silico genotoxicity and carcinogenicity prediction for food-relevant 
secondary plant metabolites.

Glück J(1), Buhrke T(2), Frenzel F(1), Braeuning A(1), Lampen A(1).

Author information:
(1)German Federal Institute for Risk Assessment, Department of Food Safety, 
Max-Dohrn-Str. 8-10, 10589 Berlin, Germany.
(2)German Federal Institute for Risk Assessment, Department of Food Safety, 
Max-Dohrn-Str. 8-10, 10589 Berlin, Germany. Electronic address: 
thorsten.buhrke@bfr.bund.de.

Humans are exposed to thousands of different secondary plant metabolites which 
may have beneficial health effects, but numerous compounds may also have toxic 
potential. In the present study we have examined the genotoxic and carcinogenic 
potential of 609 food-relevant phytochemicals by using computer models for 
toxicity prediction. We developed a scoring method and combined the results of 
different models to increase the predictive power. A combination of the VEGA 
models SARpy, KNN, ISS, and CAESAR, and of the LAZAR model "Salmonella 
typhimurium" for genotoxicity prediction performed better than the single models 
regarding specificity and accuracy. Statistical evaluation of the combined model 
for carcinogenicity prediction was not possible due to the low number of 
substances suitable for model validation. The in silico results of the present 
exercise will be useful for priority setting purposes regarding future risk 
assessment of secondary plant metabolites. Based on our analysis, 
(-)-asimilobine, aloin, annoretine, chrysothrone, coptisine, elymoclavine, and 
thalicminine were predicted to be genotoxic with high probability and may 
therefore be selected for subsequent experimental genotoxicity testing. 
Moreover, the class of pyrrolizidine alkaloids is suggested to be a high 
priority subject for further studies as these substances have been predicted to 
be carcinogenic with high probability.

Copyright © 2018 Elsevier Ltd. All rights reserved.

DOI: 10.1016/j.fct.2018.04.024
PMID: 29660365 [Indexed for MEDLINE]


5. Org Lett. 2017 Jul 7;19(13):3664-3667. doi: 10.1021/acs.orglett.7b01681. Epub 
2017 Jun 22.

Regioselective Direct C-4 Functionalization of Indole: Total Syntheses of 
(-)-Agroclavine and (-)-Elymoclavine.

Lv J(1), Wang B(1), Yuan K(1), Wang Y(1), Jia Y(1).

Author information:
(1)State Key Laboratory of Natural and Biomimetic Drugs, School of 
Pharmaceutical Sciences, Peking University , 38 Xueyuan Road, Beijing 100191, 
China.

An efficient rhodium-catalyzed method for direct C-H functionalization at the C4 
position of unprotected indoles has been developed. The utility of this method 
is demonstrated by the concise total syntheses of agroclavine and elymoclavine 
in a divergent manner. These syntheses feature a Pd-catalyzed asymmetric allylic 
alkylation reaction to assemble the triyclic indole moiety, and a ring-closing 
metathesis reaction to form the D ring.

DOI: 10.1021/acs.orglett.7b01681
PMID: 28641012