Natural gene variation inCannabis sativaunveils a key region of cannabinoid synthase enzymes

Author:

Villard CloéORCID,Bayer Christian,Medici Nora Pasquali,van de Peppel Arjen C.,Cankar KatarinaORCID,Verstappen Francel,Kappers Iris F.ORCID,Schranz M. EricORCID,Daniel BastianORCID,van Velzen RobinORCID

Abstract

AbstractCannabinoids are well-known specialised metabolites from the plantCannabis sativaL. (cannabis). They exhibit various therapeutical to intoxicating psychoactive effects and have potential for medicinal applications. Among the enzymes involved in cannabinoid biosynthesis, cannabinoid oxidocyclases such as the tetrahydrocannabinolic acid (THCA) synthase play a key role in determining cannabis chemotype. To improve our understanding of cannabinoid oxidocyclase structure-function relationship, we proposed a new approach to targeted mutagenesis. By reviewing cannabis natural variation, three cannabinoid oxidocyclase mutations (S355N, CONF, G376R) associated to atypical plant chemotypes were selected.In-vitrocharacterization of THCA synthase mutants demonstrated these mutations significantly impact enzyme activity, correlating with the associated chemotype: S355N nearly inactivated the THCA synthase, CONF impaired CBGA metabolization and altered product specificity, while G376R drastically reduced enzyme activity and altered product specificity.In-silicodocking experiments permitted to model the successive steps of THCA synthase substrate metabolization, revealing that the three mutations hamper substrate binding. Collectively, our results demonstrated how plant diversity can be leveraged to guide enzyme targeted mutagenesis, highlighted a key region of cannabinoid oxidocyclases, and permitted the establishment of a new model of the THCA synthase catalytic mechanism. This provides new insights into enzyme function, which can ultimately help developing medicinal cannabis cultivars and cannabinoid biotechnological production.

Publisher

Cold Spring Harbor Laboratory

Reference57 articles.

1. Methods for enzyme library creation: Which one will you choose?: A guide for novices and experts to introduce genetic diversity;BioEssays,2021

2. The identification of SNPs in THCA synthase gene from Pakistani Cannabis;Asia Pac J Mol Biol Biotechnol,2019

3. Clinical and Preclinical Evidence for Functional Interactions of Cannabidiol and Δ9-Tetrahydrocannabinol

4. Cannabidiol (CBD) and its analogs: a review of their effects on inflammation;Bioorg. Med. Chem,2015

5. Psychoactive constituents of cannabis and their clinical implications: a systematic review;Adicciones,2018

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3