Carotenoid Pathway Engineering in Tobacco Chloroplast Using a Synthetic Operon

Overview

Journal Mol Biotechnol

Publisher Springer

Specialties Biotechnology
Molecular Biology

Date 2023 Mar 8

PMID 36884112

Authors

Neha Tanwar

James E Rookes

David M Cahill

Sangram K Lenka

Affiliations

Soon will be listed here.

Abstract

The carotenoid pathway in plants has been altered through metabolic engineering to enhance their nutritional value and generate keto-carotenoids, which are widely sought after in the food, feed, and human health industries. In this study, the aim was to produce keto-carotenoids by manipulating the native carotenoid pathway in tobacco plants through chloroplast engineering. Transplastomic tobacco plants were generated that express a synthetic multigene operon composed of three heterologous genes, with Intercistronic Expression Elements (IEEs) for effective mRNA splicing. The metabolic changes observed in the transplastomic plants showed a significant shift towards the xanthophyll cycle, with only a minor production of keto-lutein. The use of a ketolase gene in combination with the lycopene cyclase and hydroxylase genes was a novel approach and demonstrated a successful redirection of the carotenoid pathway towards the xanthophyll cycle and the production of keto-lutein. This study presents a scalable molecular genetic platform for the development of novel keto-carotenoids in tobacco using the Design-Build-Test-Learn (DBTL) approach. This study corroborates chloroplast metabolic engineering using a synthetic biology approach for producing novel metabolites belonging to carotenoid class in industrially important tobacco plant. The synthetic multigene construct resulted in producing a novel metabolite, keto-lutein with high accumulation of xanthophyll metabolites. This figure was drawn using BioRender ( https://www.biorender.com ).

Citing Articles

Chloroplast transformation in new cultivars of tomato through particle bombardment.

Tanwar N, Mahto B, Rookes J, Cahill D, Bansal K, Lenka S 3 Biotech. 2024; 14(4):120.

PMID: 38545123 PMC: 10963679. DOI: 10.1007/s13205-024-03954-3.

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