Journal
SCIENCE BULLETIN
Volume 67, Issue 3, Pages 315-327Publisher
ELSEVIER
DOI: 10.1016/j.scib.2021.07.003
Keywords
Rational design; Carotenoid; Methylerythritol phosphate pathway (MEP); Geranylgeranyl diphosphate synthase; (GGPPS); Nicotiana tabacum
Categories
Funding
- Natural Science Foundation of Henan Province [182300410053]
- China Postdoctoral Science Foundation [2020M672308]
- Henan Postdoctoral Science Founda-tion [227462]
- Science Project [902019AA0140]
- National Key Research and Development Program of China [2019YFA0905100]
- National Natural Science Foundation of China [U2004143]
Ask authors/readers for more resources
This study demonstrates the rational engineering of NtGGPPS1 enzyme in Nicotiana tabacum, resulting in increased carotenoid production. The findings provide a basis for the structure-guided rational design of elite genes in higher plants.
Restricted genetic diversity can supply only a limited number of elite genes for modern plant cultivation and transgenesis. In this study, we demonstrate that rational design enables the engineering of geranylgeranyl diphosphate synthase (NtGGPPS), an enzyme of the methylerythritol phosphate pathway (MEP) in the model plant Nicotiana tabacum. As the crucial bottleneck in carotenoid biosynthesis, NtGGPPS1 interacts with phytoene synthase (NtPSY1) to channel GGPP into the production of carotenoids. Loss of this enzyme in the ntggpps1 mutant leads to decreased carotenoid accumulation. With the aim of enhancing NtGGPPS1 activity, we undertook structure-guided rational redesign of its substrate binding pocket in combination with sequence alignment. The activity of the designed NtGGPPS1 (a pentuple mutant of five sites V154A/I161L/F218Y/I209S/V233E, d-NtGGPPS1) was measured by a high-throughput colorimetric assay. d-NtGGPPS1 exhibited significantly higher conversion of IPP and each co-substrate (DMAPP-1995.5-fold, GPP-25.9-fold, and FPP-16.7-fold) for GGPP synthesis compared with wild type NtGGPPS1. Importantly, the transient and stable expression of d-NtGGPPS1 in the ntggpps1 mutant increased carotenoid levels in leaves, improved photosynthetic efficiency, and increased biomass relative to NtGGPPS1. These findings provide a firm basis for the engineering of GGPPS and will facilitate the development of quality and yield traits. Our results open the door for the structure-guided rational design of elite genes in higher plants. (c) 2021 Science China Press. Published by Elsevier B.V. and Science China Press. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available