Cytochrome P450 and O-methyltransferase catalyze the final steps in the biosynthesis of the anti-addictive alkaloid ibogaine from Tabernanthe iboga
| Autor: | Scott C. Farrow, Jessica Meades, Belinda Ameyaw, Mohamed O. Kamileen, Sarah E. O'Connor, Youli Xiao |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
cytochrome P450 monoterpene indole alkaloid Tabernaemontana Ibogamine Plant Biology Biochemistry Catalysis 03 medical and health sciences chemistry.chemical_compound transcriptomics Alkaloids Cytochrome P-450 Enzyme System medicine Tabernanthe iboga Secondary metabolism Molecular Biology O-methyltransferase secondary metabolism biology Apocynaceae Ibogaine Illumina sequencing Cytochrome P450 High-Throughput Nucleotide Sequencing Cell Biology ibogaine biology.organism_classification Opioid-Related Disorders Protein O-Methyltransferase natural product biosynthesis 030104 developmental biology chemistry plant biochemistry biology.protein Heterologous expression Transcriptome medicine.drug |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 1083-351X 0021-9258 |
| Popis: | Monoterpenoid indole alkaloids are a large (approximate to 3000 members) and structurally diverse class of metabolites restricted to a limited number of plant families in the order Gentianales. Tabernanthe iboga or iboga (Apocynaceae) is native to western equatorial Africa and has been used in traditional medicine for centuries. Howard Lotsof is credited with bringing iboga to the attention of Western medicine through his accidental discovery that iboga can alleviate opioid withdrawal symptoms. Since this observation, iboga has been investigated for its use in the general management of addiction. We were interested in elucidating ibogaine biosynthesis to understand the unique reaction steps en route to ibogaine. Furthermore, because ibogaine is currently sourced from plant material, these studies may help improve the ibogaine supply chain through synthetic biology approaches. Here, we used next-generation sequencing to generate the first iboga transcriptome and leveraged homology-guided gene discovery to identify the penultimate hydroxylase and final O-methyltransferase steps in ibogaine biosynthesis, herein named ibogamine 10-hydroxylase (I10H) and noribogaine-10-O-methyltransferase (N10OMT). Heterologous expression in Saccharomyces cerevisiae (I10H) or Escherichia coli (N10OMT) and incubation with putative precursors, along with HPLC-MS analysis, confirmed the predicted activities of both enzymes. Moreover, high expression levels of their transcripts were detected in ibogaine-accumulating plant tissues. These discoveries coupled with our publicly available iboga transcriptome will contribute to additional gene discovery efforts and could lead to the stabilization of the global ibogaine supply chain and to the development of ibogaine as a treatment for addiction. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|