Psychoactive Plant Database - Neuroactive Phytochemical Collection

1. Metab Eng. 2024 Sep;85:145-158. doi: 10.1016/j.ymben.2024.07.011. Epub 2024
Jul  27.

Verazine biosynthesis from simple sugars in engineered Saccharomyces cerevisiae.

Winegar PH(1), Hudson GA(1), Dell LB(2), Astolfi MCT(2), Reed J(3), Payet RD(3), 
Ombredane HCJ(3), Iavarone AT(4), Chen Y(5), Gin JW(5), Petzold CJ(5), Osbourn 
AE(3), Keasling JD(6).

Author information:
(1)Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, Emeryville, 
CA, 94608, USA; Biological Systems and Engineering, Lawrence Berkeley National 
Laboratory, Berkeley, CA, 94720, USA; California Institute for Quantitative 
Biosciences (QB3 Institute), University of California, Berkeley, CA, 94720, USA.
(2)Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, Emeryville, 
CA, 94608, USA; Biological Systems and Engineering, Lawrence Berkeley National 
Laboratory, Berkeley, CA, 94720, USA; Department of Chemical and Biomolecular 
Engineering and Department of Bioengineering, University of California, 
Berkeley, CA, 94720, USA.
(3)John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, UK.
(4)California Institute for Quantitative Biosciences (QB3 Institute), University 
of California, Berkeley, CA, 94720, USA.
(5)Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, Emeryville, 
CA, 94608, USA; Biological Systems and Engineering, Lawrence Berkeley National 
Laboratory, Berkeley, CA, 94720, USA.
(6)Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, Emeryville, 
CA, 94608, USA; Biological Systems and Engineering, Lawrence Berkeley National 
Laboratory, Berkeley, CA, 94720, USA; California Institute for Quantitative 
Biosciences (QB3 Institute), University of California, Berkeley, CA, 94720, USA; 
Department of Chemical and Biomolecular Engineering and Department of 
Bioengineering, University of California, Berkeley, CA, 94720, USA; The Novo 
Nordisk Foundation Center for Biosustainability, Technical University Denmark, 
Kemitorvet, Building 220, Kongens, Lyngby, 2800, Denmark. Electronic address: 
keasling@berkeley.edu.

Steroidal alkaloids are FDA-approved drugs (e.g., Zytiga) and promising drug 
candidates/leads (e.g., cyclopamine); yet many of the ≥697 known steroidal 
alkaloid natural products remain underutilized as drugs because it can be 
challenging to scale their biosynthesis in their producing organisms. 
Cyclopamine is a steroidal alkaloid produced by corn lily (Veratrum spp.) 
plants, and it is an inhibitor of the Hedgehog (Hh) signaling pathway. 
Therefore, cyclopamine is an important drug candidate/lead to treat human 
diseases that are associated with dysregulated Hh signaling, such as basal cell 
carcinoma and acute myeloid leukemia. Cyclopamine and its semi-synthetic 
derivatives have been studied in (pre)clinical trials as Hh inhibitor-based 
drugs. However, challenges in scaling the production of cyclopamine have slowed 
efforts to improve its efficacy and safety profile through (bio)synthetic 
derivatization, often limiting drug development to synthetic analogs of 
cyclopamine such as the FDA-approved drugs Odomzo, Daurismo, and Erivedge. If a 
platform for the scalable and sustainable production of cyclopamine were 
established, then its (bio)synthetic derivatization, clinical development, and, 
ultimately, widespread distribution could be accelerated. Ongoing efforts to 
achieve this goal include the biosynthesis of cyclopamine in Veratrum plant cell 
culture and the semi-/total chemical synthesis of cyclopamine. Herein, this work 
advances efforts towards a promising future approach: the biosynthesis of 
cyclopamine in engineered microorganisms. We completed the heterologous 
microbial production of verazine (biosynthetic precursor to cyclopamine) from 
simple sugars (i.e., glucose and galactose) in engineered Saccharomyces 
cerevisiae (S. cerevisiae) through the inducible upregulation of the native 
yeast mevalonate and lanosterol biosynthetic pathways, diversion of biosynthetic 
flux from ergosterol (i.e., native sterol in S. cerevisiae) to cholesterol 
(i.e., biosynthetic precursor to verazine), and expression of a refactored 
five-step verazine biosynthetic pathway. The engineered S. cerevisiae strain 
that produced verazine contains eight heterologous enzymes sourced from seven 
different species. Importantly, S. cerevisiae-produced verazine was 
indistinguishable via liquid chromatography-mass spectrometry from both a 
commercial standard (Veratrum spp. plant-produced) and Nicotiana 
benthamiana-produced verazine. To the best of our knowledge, this is the first 
report describing the heterologous production of a steroidal alkaloid in an 
engineered yeast. Verazine production was ultimately increased through 
design-build-test-learn cycles to a final titer of 83 ± 3 μg/L (4.1 ± 0.1 μg/g 
DCW). Together, this research lays the groundwork for future microbial 
biosynthesis of cyclopamine, (bio)synthetic derivatives of cyclopamine, and 
other steroidal alkaloid natural products.

Copyright © 2024 International Metabolic Engineering Society. Published by 
Elsevier Inc. All rights reserved.

DOI: 10.1016/j.ymben.2024.07.011
PMCID: PMC11421371
PMID: 39074544 [Indexed for MEDLINE]

Conflict of interest statement: Declaration of interests J.D.K. has a financial 
interest in Demetrix, Maple Bio, Lygos, Napigen, Berkeley Yeast, Zero Acre 
Farms, Ansa Biotechnologies, Apertor Pharmaceuticals, BioMia, ResVit Bio, and 
Cyklos Materials. All other authors declare no competing interests.


2. Plant Commun. 2024 Jun 10;5(6):100831. doi: 10.1016/j.xplc.2024.100831. Epub 
2024 Feb 1.

Efficient heterologous biosynthesis of verazine, a metabolic precursor of the 
anti-cancer drug cyclopamine, in Nicotiana benthamiana.

Kou C(1), Liu J(2), Yin X(3), He D(2), Liu J(3), Hua X(3), Ma R(4), Sun W(5), 
Xue Z(6), Ma P(7).

Author information:
(1)College of Life Sciences, Northwest A&F University, Yangling 712100, China; 
Key Laboratory of Saline-alkali Vegetation Ecology Restoration (Northeast 
Forestry University), Ministry of Education, Harbin 150040, China; Heilongjiang 
Key Laboratory of Plant Bioactive Substance Biosynthesis and Utilization, 
Northeast Forestry University, Harbin 150040, China.
(2)College of Life Sciences, Northwest A&F University, Yangling 712100, China.
(3)Key Laboratory of Saline-alkali Vegetation Ecology Restoration (Northeast 
Forestry University), Ministry of Education, Harbin 150040, China; Heilongjiang 
Key Laboratory of Plant Bioactive Substance Biosynthesis and Utilization, 
Northeast Forestry University, Harbin 150040, China.
(4)Jilin Provincial Key Laboratory of Agricultural Biotechnology, Jilin Academy 
of Agricultural Sciences, Changchun 130033, China.
(5)Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese 
Medicine, Institute of Chinese Materia Medica, China Academy of Chinese Medical 
Sciences, Beijing 100700, China.
(6)Key Laboratory of Saline-alkali Vegetation Ecology Restoration (Northeast 
Forestry University), Ministry of Education, Harbin 150040, China; Heilongjiang 
Key Laboratory of Plant Bioactive Substance Biosynthesis and Utilization, 
Northeast Forestry University, Harbin 150040, China. Electronic address: 
xuezhy@126.com.
(7)College of Life Sciences, Northwest A&F University, Yangling 712100, China. 
Electronic address: mapengda@163.com.

DOI: 10.1016/j.xplc.2024.100831
PMCID: PMC11211220
PMID: 38308438 [Indexed for MEDLINE]


3. Molecules. 2023 Oct 16;28(20):7116. doi: 10.3390/molecules28207116.

Steroidal Alkaloids from the Roots of Veratrum mengtzeanum Loes. with Their 
Anti-Inflammatory Activities.

Yuan W(1)(2), Ma J(2)(3), Liu X(2)(3), Zi C(1)(2), Xi Y(1)(2), Shen X(1), Li 
G(4), Sheng J(2), Wang X(2).

Author information:
(1)College of Science, Yunnan Agricultural University, Kunming 650201, China.
(2)Key Laboratory of Pu'er Tea, Ministry of Education, Yunnan Agricultural 
University, Kunming 650201, China.
(3)College of Food Science and Technology, Yunnan Agricultural University, 
Kunming 650201, China.
(4)College of Chinese Medicine, Yunnan University of Chinese Medicine, Kunming 
650500, China.

The phytochemical investigation of Veratrum mengtzeanum Loes. roots resulted in 
the isolation and characterization of two novel, namely Mengtzeanines A (1), 
Mengtzeanines B (2), and eight known steroidal alkaloids (3-10). Their 
structural properties were assessed though extensive spectroscopic techniques. 
All constituents 1-10 were analyzed for suppression of NO formation in 
LPS-induced RAW264.7 macrophages. Among them, constituent 6 (Verazine) showed 
inhibition against LPS-induced NO production (IC50 = 20.41 μM). Additionally, 
compound 6 could inhibit the secretion of IL1β, IL6, and TNFα, and downregulate 
the productions of iNOS and COX2 in LPS-induced RAW264.7 macrophages. Further 
experiments revealed that 6 exhibited a potent anti-inflammatory level in 
LPS-stimulated RAW264.7 macrophages via inhibiting NF-κB, and triggering of 
Keap1/Nrf2/HO-1 axis, implying that compound 6 may be a promising candidate for 
treating inflammatory disorders.

DOI: 10.3390/molecules28207116
PMCID: PMC10609212
PMID: 37894597 [Indexed for MEDLINE]

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


4. Phytochemistry. 2023 Sep;213:113768. doi: 10.1016/j.phytochem.2023.113768.
Epub  2023 Jun 19.

Undescribed steroidal alkaloids from the bulbs of Fritillaria sinica.

Li JY(1), Wu SF(1), An YL(2), Yao CL(1), Yao S(1), Huang Y(1), Wei WL(1), Zhang 
JQ(1), Bi QR(1), Qu H(1), Guo DA(3).

Author information:
(1)National Engineering Research Center of TCM Standardization Technology, 
Shanghai Research Center for Modernization of Traditional Chinese Medicine, 
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 
201203, China.
(2)National Engineering Research Center of TCM Standardization Technology, 
Shanghai Research Center for Modernization of Traditional Chinese Medicine, 
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 
201203, China; University of Chinese Academy of Sciences, Beijing, 100049, 
China.
(3)National Engineering Research Center of TCM Standardization Technology, 
Shanghai Research Center for Modernization of Traditional Chinese Medicine, 
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 
201203, China; University of Chinese Academy of Sciences, Beijing, 100049, 
China. Electronic address: daguo@simm.ac.cn.

Eight undescribed steroidal alkaloid derivatives, including three cevanine-type 
isosteroidal alkaloids (two N-oxide glycosides and one D-ring aromatization) 
(1-3), one verazine-type steroidal alkaloid derivative (4), three 
solanidine-type steroidal alkaloid glycosides (5-7), and one veratramine-type 
analogue (8), along with three known compounds (9-11) were isolated from the 
bulbs of Fritillaria sinica. Their structures were elucidated by comprehensive 
analysis of spectroscopic data, acidic hydrolysis, and X-ray crystal 
diffractions. In the in vitro bioassay, the anti-cancer effect, anti-oxidation 
and anti-inflammatory activities for the isolates were evaluated at a 
concentration of 10 μM.

Copyright © 2023 Elsevier Ltd. All rights reserved.

DOI: 10.1016/j.phytochem.2023.113768
PMID: 37343737 [Indexed for MEDLINE]

Conflict of interest statement: Declaration of competing interest The authors 
declare that they have no known competing financial interests or personal 
relationships that could have appeared to influence the work reported in this 
paper.


5. Biotechnol Genet Eng Rev. 2023 Oct;39(2):760-774. doi: 
10.1080/02648725.2022.2162264. Epub 2022 Dec 28.

In silico toxicity prediction, molecular docking studies and in vitro validation 
of antibacterial potential of alkaloids from Eclipta alba in designing of novel 
antimicrobial therapeutic strategies.

Ameen F(1), Orfali R(2), Mamidala E(3), Davella R(3).

Author information:
(1)Department of Botany & Microbiology, College of Science, King Saud 
University, Riyadh, Saudi Arabia.
(2)Department of Pharmacognosy, College of Pharmacy, King Saud University, 
Riyadh, Saudi Arabia.
(3)Department of Zoology, Infectious Diseases Research Lab, Kakatiya University, 
Warangal, India.

The rapid emergence of various drug resistance and unfavourable aliphatic 
medication side effects endangers people's health. Phytocompounds with 
antibacterial activity and less harmful effects are known to be present in 
medicinal plants. Alkaloids from Eclipta alba were tested for their in vitro 
antibacterial capabilities and in silico docking studies against pathogenic 
bacteria and their target proteins in the current investigation. The alkaloid 
compounds verazine, ecliptine, 4-hydroxyverazine, 20-Epi-4beta-hydroxyverazine 
and hydroxyverazine were subjected to molecular docking studies to determine the 
method of binding as well as potential interactions and the docking score. The 
in vitro antibacterial activity of verazine alkaloid was assessed against two 
gram-positive and two gram-negative bacteria. Verazine alkaloid has the best 
inhibitory ability against DNA gyrase of E. coli (ΔG= -8.44 kcal/mol) and 
dihydrofolate reductase (DHFR) of S. aureus (ΔG= -10.04 kcal/mol), according to 
docking studies. Verazine shown substantial in vitro antibacterial activity in 
this investigation against all test bacteria, with MIC and MBC values of 31.25 
and 62.50 µg/mL for S. aureus and 15.63 and 31.25 µg/mL for B. cereus, 
respectively. The results of this work highlighted the value of unique alkaloid 
compounds from E. alba, which may offer effective antibacterial agents and DNA 
gyrase, DHFR inhibitors due to their novel structural properties capable of 
combating antimicrobial resistance. These findings call for more investigation 
into the compounds' function as antibacterial agents, as well as their 
unique-binding locations and mechanisms.

DOI: 10.1080/02648725.2022.2162264
PMID: 36578142 [Indexed for MEDLINE]