Enhancing taxane biosynthesis in cell suspension culture of Taxus chinensis by overexpressing the neutral/alkaline invertase gene
Enhancing taxane biosynthesis in cell suspension culture of Taxus chinensis by overexpressing the neutral/alkaline invertase gene
复制标题
通过过表达中性/碱性转化酶基因增强红豆杉细胞悬浮培养中紫杉烷的生物合成
DOI：
10.1016/j.procbio.2015.01.018
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发表时间：
2015-04-01
影响因子：
4.4
通讯作者：
Yu, Longjiang
中科院分区：
文献类型：
--
作者：
Dong, Yanshan;Duan, Wenlan;Yu, Longjiang
蔗糖代谢产生物质和能量基础并调节植物细胞培养物中次生代谢物的生物合成。然而，这种调节机制尚不清楚。中性/碱性转化酶（NINV）基因是蔗糖水解的关键酶之一。由此，从红豆杉细胞中获得了该基因的完整开放阅读框（ORF）。该基因也在大肠杆菌中成功表达。通过蔗糖裂解功能验证了表达的蛋白。此外，NINV基因的转录水平可以响应蔗糖浓度的变化，并与紫杉烷细胞中紫杉烷的生物合成有关。随后，我们开发了基因表达载体并将其导入紫杉悬浮细胞中，以证实NINV基因在紫杉烷生物合成中的作用。结果表明，该基因过度表达，并且显着增强了紫杉二烯合酶（TAS）基因的表达，以及检测到的七种紫杉烷的生物合成。因此，NINV介导的蔗糖代谢可以促进次级代谢产物的生物合成。这项研究提出了增加植物细胞培养物中次生代谢产物产量的新概念和方法。 (C) 2015 Elsevier Ltd. 保留所有权利。
Sucrose metabolism generates material and energy foundations and regulates the biosynthesis of secondary metabolites in plant cell cultures. However, this regulation mechanism is unclear. The neutral/alkaline invertase (NINV) gene is among the key enzymes in sucrose hydrolysis. Thus, the complete open reading frame (ORF) of this gene was obtained from Taxus chinensis cells. This gene was also successfully expressed in Escherichia coli. The expressed protein was verified through the function of sucrose cleavage. Furthermore, the transcriptional levels of the NINV gene can respond to the sucrose concentration changes and relate to taxane biosynthesis in T. chinensis cells. Subsequently, we developed the gene expression vector and introduced it into T. chinensis suspension cells to confirm the role of the NINV gene in taxane biosynthesis. Results indicated that this gene was overexpressed and that it significantly enhanced the expression of the taxadiene synthase (TAS) gene, as well as the biosynthesis of the seven detected individual taxanes. Thus, NINV-mediated sucrose metabolism can promote the biosynthesis of secondary metabolites. This study presents new concepts and approaches to increase the production of secondary metabolites in plant cell cultures. (C) 2015 Elsevier Ltd. All rights reserved.