Engineering and Optimization of the Salidroside Production in Tobacco Cells by Reconstructed Biosynthetic Genes from Rhodiola rosea.

IF 3.7 2区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

ACS Synthetic Biology Pub Date : 2025-07-18 Epub Date: 2025-06-24 DOI:10.1021/acssynbio.5c00185

Huapeng Sun, Xin Tang, Yingying Zhang, Xiaona Fu, Rongxiang Wang, Muhammad Shahzaib, Fei Qiao

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引用次数: 0

Abstract

Salidroside, a tyrosine-derived bioactive product, originates from the Rhodiola genus and has various medicinal properties. However, the surge in global demand requires exploration of more efficient and sustainable approaches to produce salidroside. This study reconstructed three functional enzymes in tobacco cells to produce salidroside using endogenous l-tyrosine. Under the optimized two-stage protocol (1% sucrose in the production medium; 11 days of culture), salidroside accumulation reached a maximum of 377.53 μg per gram of HHU cells in fresh weight. Neither precursor feeding (100 mg/L shikimate) nor various elicitors (e.g., MeJA, SA, and ABA) provided any further enhancement. Comparative transcriptomic analysis revealed that two DAHPS transcripts (107797958 and 107817203) were upregulated more than 2-fold, indicating that the reconstructed pathway boosts endogenous l-tyrosine flux, and elicitation and precursor feeding strategies could not enhance salidroside accumulation. Meanwhile, comparative transcriptomic analysis revealed that the reconstructed pathway could upregulate crucial gene expression in endogenous l-tyrosine biosynthesis. This study evaluates tobacco cells as biofactories to produce salidroside and lays the foundation for the biosynthesis engineering of other heterologous metabolites.

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重建红景天生物合成基因在烟草细胞中生产红景天苷的工程与优化。

红景天苷是一种酪氨酸衍生的生物活性产品，起源于红景天属，具有多种药用特性。然而，全球需求的激增要求探索更有效和可持续的方法来生产红景天苷。本研究利用内源性l-酪氨酸在烟草细胞中重建了三种功能酶以生产红景天苷。在优化的两阶段方案下(生产培养基中添加1%蔗糖；培养11 d时，红景天苷积累量最高，为377.53 μg / g / g。前体饲喂（100 mg/L莽草酸）和各种激发子（如MeJA， SA和ABA）均未进一步增强。比较转录组学分析显示，两个DAHPS转录本（107797958和107817203）上调了2倍以上，表明重建途径促进了内源性l-酪氨酸通量，诱导和前体取食策略不能增强红豆杉苷的积累。同时，比较转录组学分析显示，重建的途径可以上调内源性l-酪氨酸生物合成中关键基因的表达。本研究评价了烟草细胞作为生物工厂生产红萝卜苷，为其他异源代谢物的生物合成工程奠定了基础。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

ACS Synthetic Biology 生物-

CiteScore

8.00

自引率

10.60%

发文量

380

审稿时长

6-12 weeks

期刊介绍： The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism. Topics may include, but are not limited to: Design and optimization of genetic systems Genetic circuit design and their principles for their organization into programs Computational methods to aid the design of genetic systems Experimental methods to quantify genetic parts, circuits, and metabolic fluxes Genetic parts libraries: their creation, analysis, and ontological representation Protein engineering including computational design Metabolic engineering and cellular manufacturing, including biomass conversion Natural product access, engineering, and production Creative and innovative applications of cellular programming Medical applications, tissue engineering, and the programming of therapeutic cells Minimal cell design and construction Genomics and genome replacement strategies Viral engineering Automated and robotic assembly platforms for synthetic biology DNA synthesis methodologies Metagenomics and synthetic metagenomic analysis Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction Gene optimization Methods for genome-scale measurements of transcription and metabolomics Systems biology and methods to integrate multiple data sources in vitro and cell-free synthetic biology and molecular programming Nucleic acid engineering.