{"title":"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","authors":"Wei Song, Tong Li, Shan Yan, Mingyue Zhang, Xiaojing Ma, Liping Kang, Xin Hua, Zheyong Xue","doi":"10.1111/tpj.70237","DOIUrl":null,"url":null,"abstract":"<div>\n \n <p>Polyphyllins, a prominent class of steroidal saponins in <i>Paris</i> 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-<span>l</span>-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-<span>d</span>-Glc and UDP-<span>d</span>-Xyl. Furthermore, co-expression of <i>PpUGT</i> genes with upstream biosynthetic genes in <i>Nicotiana benthamiana</i> enabled <i>de novo</i> synthesis of polyphyllins III and II, achieving yields of 93.64 and 68.39 μg g<sup>−1</sup> 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.</p>\n </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"122 4","pages":""},"PeriodicalIF":5.7000,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Two rhamnosyltransferases for de novo biosynthesis of polyphyllins in Nicotiana benthamiana\",\"authors\":\"Wei Song, Tong Li, Shan Yan, Mingyue Zhang, Xiaojing Ma, Liping Kang, Xin Hua, Zheyong Xue\",\"doi\":\"10.1111/tpj.70237\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n <p>Polyphyllins, a prominent class of steroidal saponins in <i>Paris</i> 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-<span>l</span>-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-<span>d</span>-Glc and UDP-<span>d</span>-Xyl. Furthermore, co-expression of <i>PpUGT</i> genes with upstream biosynthetic genes in <i>Nicotiana benthamiana</i> enabled <i>de novo</i> synthesis of polyphyllins III and II, achieving yields of 93.64 and 68.39 μg g<sup>−1</sup> 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.</p>\\n </div>\",\"PeriodicalId\":233,\"journal\":{\"name\":\"The Plant Journal\",\"volume\":\"122 4\",\"pages\":\"\"},\"PeriodicalIF\":5.7000,\"publicationDate\":\"2025-05-27\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"The Plant Journal\",\"FirstCategoryId\":\"2\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1111/tpj.70237\",\"RegionNum\":1,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"PLANT SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"The Plant Journal","FirstCategoryId":"2","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/tpj.70237","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
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.
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.