两种鼠李糖转移酶在烟叶中用于多叶磷脂的新生生物合成

IF 5.7 1区 生物学 Q1 PLANT SCIENCES
Wei Song, Tong Li, Shan Yan, Mingyue Zhang, Xiaojing Ma, Liping Kang, Xin Hua, Zheyong Xue
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引用次数: 0

摘要

多叶磷脂是巴黎植物中一类突出的甾体皂素,其多样的生物活性归功于其糖单元构型,尽管对其糖基化途径的了解甚少。在这里,我们利用异源表达系统鉴定并表征了两种udp -鼠李糖转移酶,PpUGT73YD1和PpUGT738A2。这些酶依次催化多叶树素V和VI转化为三糖和四糖衍生物。PpUGT73YD1可同时识别螺皮苷和呋喃皂苷,而PpUGT738A2可特异性识别螺皮苷。这两种酶都对作为糖供体的UDP-l-Rha具有严格的特异性。PpUGT73YD1的结构建模和定点突变显示,T149M和L283A突变使糖供体偏好向UDP-d-Glc和UDP-d-Xyl转移。此外,pppugt基因与上游生物合成基因在烟叶中共表达,使多叶磷脂III和II重新合成,叶片产量分别为93.64和68.39 μg−1干重。本研究阐明了两个关键的鼠李糖转移酶在多叶毛素生物合成中的作用,并通过工程植物底盘证明了它们参与甾体皂苷的生产。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Two rhamnosyltransferases for de novo biosynthesis of polyphyllins in Nicotiana benthamiana

Two rhamnosyltransferases for de novo biosynthesis of polyphyllins in Nicotiana benthamiana

Polyphyllins, a prominent class of steroidal saponins in Paris species, owe their diverse bioactivities to their sugar unit configurations, though their glycosylation pathways remain poorly understood. Here, we identified and characterized two UDP-rhamnosyltransferases, PpUGT73YD1 and PpUGT738A2, using heterologous expression systems. These enzymes sequentially catalyze the conversion of polyphyllin V and VI into trisaccharide and tetrasaccharide derivatives, respectively. While PpUGT73YD1 accommodates both spiro and furo saponins, PpUGT738A2 specifically recognizes spiro saponins. Both enzymes exhibit strict specificity for UDP-l-Rha as a sugar donor. Structural modeling and site-directed mutagenesis of PpUGT73YD1 revealed that mutations at T149M and L283A shifted sugar donor preference toward UDP-d-Glc and UDP-d-Xyl. Furthermore, co-expression of PpUGT genes with upstream biosynthetic genes in Nicotiana benthamiana enabled de novo synthesis of polyphyllins III and II, achieving yields of 93.64 and 68.39 μg g−1 dry weight in leaves, respectively. This study elucidates the roles of two key rhamnosyltransferases in polyphyllin biosynthesis and demonstrates their involvement in steroidal saponin production through an engineered plant chassis.

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来源期刊
The Plant Journal
The Plant Journal 生物-植物科学
CiteScore
13.10
自引率
4.20%
发文量
415
审稿时长
2.3 months
期刊介绍: Publishing the best original research papers in all key areas of modern plant biology from the world"s leading laboratories, The Plant Journal provides a dynamic forum for this ever growing international research community. Plant science research is now at the forefront of research in the biological sciences, with breakthroughs in our understanding of fundamental processes in plants matching those in other organisms. The impact of molecular genetics and the availability of model and crop species can be seen in all aspects of plant biology. For publication in The Plant Journal the research must provide a highly significant new contribution to our understanding of plants and be of general interest to the plant science community.
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