ArticleJournal of biological engineering2015
Heterologous expression of the isopimaric acid pathway in Nicotiana benthamiana and the effect of N-terminal modifications of the involved cytochrome P450 enzyme.
Article in Journal of biological engineering, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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24 citing papers in PubMed, 40 citations in OpenAlex.
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- Divergent multifunctional P450s-empowered biosynthesis of bioactive tripterifordin and cryptic atiserenoids in Aconitum implies convergent evolution.Nature communications · 2025Article
- Thylakoid Targeting Improves Stability of a Cytochrome P450 in the CyanobacteriumACS synthetic biology · 2025Article
- Strangers in a foreign land: 'Yeastizing' plant enzymes.Microbial biotechnology · 2024Review
- Miltiradiene Production by Cytoplasmic Metabolic Engineering inMetabolites · 2023Article
- An improved Nicotiana benthamiana bioproduction chassis provides novel insights into nicotine biosynthesis.The New phytologist · 2023Article
- Directing cyanobacterial photosynthesis in a cytochrome c oxidase mutant using a heterologous electron sink.Plant physiology · 2022Article
- Evaluation of biotransformation capacity of transplastomic plants and hairy roots of Nicotiana tabacum expressing human cytochrome P450 2D6.Transgenic research · 2022Article
- Exploiting photosynthesis-driven P450 activity to produce indican in tobacco chloroplasts.Frontiers in plant science · 2022Article
- Combinatorial assembly platform enabling engineering of genetically stable metabolic pathways in cyanobacteria.Nucleic acids research · 2021Article
- Evolution-aided engineering of plant specialized metabolism.aBIOTECH · 2021Review
- Structural and functional analysis of tomato sterol C22 desaturase.BMC plant biology · 2021Article
- Introduction of a green algal squalene synthase enhances squalene accumulation in a strain ofMetabolic engineering communications · 2020Article
- A force awakens: exploiting solar energy beyond photosynthesis.Journal of experimental botany · 2019Review
- Cytosolic lipid droplets as engineered organelles for production and accumulation of terpenoid biomaterials in leaves.Nature communications · 2019Article
- Article
- Review
- Engineering terpenoid production through transient expression in Nicotiana benthamiana.Plant cell reports · 2018Review
- Photosynthetic fuel for heterologous enzymes: the role of electron carrier proteins.Photosynthesis research · 2017Review
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundPlant terpenoids are known for their diversity, stereochemical complexity, and their commercial interest as pharmaceuticals, food additives, and cosmetics. Developing biotechnology approaches for the production of these compounds in heterologous hosts can increase their market availability, reduce their cost, and provide sustainable production platforms. In this context, we aimed at producing the antimicrobial diterpenoid isopimaric acid from Sitka spruce. Isopimaric acid is synthesized using geranylgeranyl diphosphate as a precursor molecule that is cyclized by a diterpene synthase in the chloroplast and subsequently oxidized by a cytochrome P450, CYP720B4.
resultsWe transiently expressed the isopimaric acid pathway in Nicotiana benthamiana leaves and enhanced its productivity by the expression of two rate-limiting steps in the pathway (providing the general precursor of diterpenes). This co-expression resulted in 3-fold increase in the accumulation of both isopimaradiene and isopimaric acid detected using GC-MS and LC-MS methodology. We also showed that modifying or deleting the transmembrane helix of CYP720B4 does not alter the enzyme activity and led to successful accumulation of isopimaric acid in the infiltrated leaves. Furthermore, we demonstrated that a modified membrane anchor is a prerequisite for a functional CYP720B4 enzyme when the chloroplast targeting peptide is added. We report the accumulation of 45-55 μg/g plant dry weight of isopimaric acid four days after the infiltration with the modified enzymes.
conclusionsIt is possible to localize a diterpenoid pathway from spruce fully within the chloroplast of N. benthamiana and a few modifications of the N-terminal sequences of the CYP720B4 can facilitate the expression of plant P450s in the plastids. The coupling of terpene biosynthesis closer to photosynthesis paves the way for light-driven biosynthesis of valuable terpenoids.
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