Rational design of geranylgeranyl diphosphate synthase enhances carotenoid production and improves photosynthetic efficiency in Nicotiana tabacum

作     者：Chen Dong Ge Qu Jinggong Guo Fang Wei Shuwen Gao Zhoutong Sun Lifeng Jin Xuwu Sun Jean-David Rochaix Yuchen Miao Ran Wang 董臣;曲戈;郭敬功;位芳;高书文;孙周通;金立锋;孙旭武;Jean-David Rochaix;苗雨晨;王燃

作者机构：Zhengzhou Tobacco Research InstituteZhengzhou 450001China Key Laboratory of Plant Stress BiologyState Key Laboratory of Cotton BiologySchool of Life SciencesHenan UniversityKaifeng 475001China College of Biological EngineeringHenan University of TechnologyZhengzhou 450001China Tianjin Institute of Industrial BiotechnologyChinese Academy of SciencesTianjin 300308China School of Agricultural SciencesZhengzhou UniversityZhengzhou 450001China Departments of Molecular Biology and Plant BiologyUniversity of GenevaGeneva 1211Switzerland 

出 版 物：《Science Bulletin》 (科学通报（英文版）)

年 卷 期：2022年第67卷第3期

页      面：315-327页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 090102[农学-作物遗传育种] 

基　　金：the Natural Science Foundation of Henan Province(182300410053) the China Postdoctoral Science Foundation(2020M672308) Henan Postdoctoral Science Foundation(227462) Science Project(902019AA0140) the National Key Research and Development Program of China(2019YFA0905100) the National Natural Science Foundation of China(U2004143) 

主　　题：Rational design Carotenoid Methylerythritol phosphate pathway(MEP) Geranylgeranyl diphosphate synthase(GGPPS) Nicotiana tabacum 

摘      要：Restricted genetic diversity can supply only a limited number of elite genes for modern plant cultivation and *** this study,we demonstrate that rational design enables the engineering of geranyl-geranyl diphosphate synthase(NtGGPPS),an enzyme of the methylerythritol phosphate pathway(MEP)in the model plant Nicotiana *** the crucial bottleneck in carotenoid biosynthesis,NtGGPPS1 interacts with phytoene synthase(NtPSY1)to channel GGPP into the production of *** of this enzyme in the ntggpps1 mutant leads to decreased carotenoid *** the aim of enhanc-ing NtGGPPS1 activity,we undertook structure-guided rational redesign of its substrate binding pocket in combination with sequence *** 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 ***,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 *** findings provide a firm basis for the engineering of GGPPS and will facilitate the development of quality and yield *** results open the door for the structure-guided rational design of elite genes in higher plants。