Mohamed O. Kamileen, Yoko Nakamura, Katrin Luck, Sarah Heinicke, Benke Hong, Maite Colinas, Benjamin R. Lichman, Sarah E. O'Connor
{"title":"Streamlined screening platforms lead to the discovery of pachysiphine synthase from Tabernanthe iboga","authors":"Mohamed O. Kamileen, Yoko Nakamura, Katrin Luck, Sarah Heinicke, Benke Hong, Maite Colinas, Benjamin R. Lichman, Sarah E. O'Connor","doi":"10.1111/nph.20133","DOIUrl":null,"url":null,"abstract":"Summary<jats:list list-type=\"bullet\"> <jats:list-item>Plant‐specialized metabolism is largely driven by the oxidative tailoring of key chemical scaffolds catalyzed by cytochrome P450 (CYP450s) enzymes. Monoterpene indole alkaloids (MIAs) tabersonine and pseudo‐tabersonine, found in the medicinal plant <jats:italic>Tabernanthe iboga</jats:italic> (commonly known as iboga), are tailored with oxidations, and the enzymes involved remain unknown.</jats:list-item> <jats:list-item>Here, we developed a streamlined screening strategy to test the activity of <jats:italic>T. iboga</jats:italic> CYP450s in <jats:italic>Nicotiana benthamiana</jats:italic>. Using multigene constructs encoding the biosynthesis of tabersonine and pseudo‐tabersonine scaffolds, we aimed to uncover the CYP450s responsible for oxidative transformations in these scaffolds.</jats:list-item> <jats:list-item>Our approach identified two <jats:italic>T. iboga</jats:italic> cytochrome P450 enzymes: pachysiphine synthase (PS) and 16‐hydroxy‐tabersonine synthase (T16H). These enzymes catalyze an epoxidation and site‐specific hydroxylation of tabersonine to produce pachysiphine and 16‐OH‐tabersonine, respectively.</jats:list-item> <jats:list-item>This work provides new insights into the biosynthetic pathways of MIAs and underscores the utility of <jats:italic>N. benthamiana</jats:italic> and <jats:italic>Catharanthus roseus</jats:italic> as platforms for the functional characterization of plant enzymes.</jats:list-item> </jats:list>","PeriodicalId":214,"journal":{"name":"New Phytologist","volume":null,"pages":null},"PeriodicalIF":8.3000,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"New Phytologist","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1111/nph.20133","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
引用次数: 0
Abstract
SummaryPlant‐specialized metabolism is largely driven by the oxidative tailoring of key chemical scaffolds catalyzed by cytochrome P450 (CYP450s) enzymes. Monoterpene indole alkaloids (MIAs) tabersonine and pseudo‐tabersonine, found in the medicinal plant Tabernanthe iboga (commonly known as iboga), are tailored with oxidations, and the enzymes involved remain unknown.Here, we developed a streamlined screening strategy to test the activity of T. iboga CYP450s in Nicotiana benthamiana. Using multigene constructs encoding the biosynthesis of tabersonine and pseudo‐tabersonine scaffolds, we aimed to uncover the CYP450s responsible for oxidative transformations in these scaffolds.Our approach identified two T. iboga cytochrome P450 enzymes: pachysiphine synthase (PS) and 16‐hydroxy‐tabersonine synthase (T16H). These enzymes catalyze an epoxidation and site‐specific hydroxylation of tabersonine to produce pachysiphine and 16‐OH‐tabersonine, respectively.This work provides new insights into the biosynthetic pathways of MIAs and underscores the utility of N. benthamiana and Catharanthus roseus as platforms for the functional characterization of plant enzymes.
期刊介绍:
New Phytologist is an international electronic journal published 24 times a year. It is owned by the New Phytologist Foundation, a non-profit-making charitable organization dedicated to promoting plant science. The journal publishes excellent, novel, rigorous, and timely research and scholarship in plant science and its applications. The articles cover topics in five sections: Physiology & Development, Environment, Interaction, Evolution, and Transformative Plant Biotechnology. These sections encompass intracellular processes, global environmental change, and encourage cross-disciplinary approaches. The journal recognizes the use of techniques from molecular and cell biology, functional genomics, modeling, and system-based approaches in plant science. Abstracting and Indexing Information for New Phytologist includes Academic Search, AgBiotech News & Information, Agroforestry Abstracts, Biochemistry & Biophysics Citation Index, Botanical Pesticides, CAB Abstracts®, Environment Index, Global Health, and Plant Breeding Abstracts, and others.