<br>人细胞色素 P450s 2D6 和 3A4 在烟草中的瞬时表达为快速底物测试和新型化合物的生产提供了可能性,Biotechnology Journal

当前位置： X-MOL 学术 › Biotechnol. J. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

人细胞色素 P450s 2D6 和 3A4 在烟草中的瞬时表达为快速底物测试和新型化合物的生产提供了可能性
Biotechnology Journal ( IF 3.2 ) Pub Date : 2018-04-26 , DOI: 10.1002/biot.201700696
Yuriy V Sheludko _{1,

2} , Iryna M Gerasymenko _{1,

2} , Heribert Warzecha ₁

Affiliation

利用外源基因的瞬时表达来生物转化具有药用价值的低分子量化合物（包括植物次生代谢物）是植物生物技术中仍很少探索的一个诱人趋势。在目前的工作中，提出了一种有效的方案，用于通过农杆菌介导的本塞姆塞姆氏烟草中的瞬时表达来快速评估作为人类 P450（CYP2D6 和 CYP3A4）潜在底物的合成和植物衍生代谢物。具有广泛底物特异性的动物 P450 是转化多种代谢物的有希望的候选者。 P450 在异源环境中的效率并不总是令人满意，并且取决于相关电子转移酶的可用性。植物代表了外源 P450 表达的一系列有吸引力的潜在宿主。设计了提供最高瞬时表达效率的最佳遗传块组成，验证了有效的底物施用方案，并测试了所研究的 P450 对氯雷他定和具有不同分子支架结构的几种吲哚生物碱的生物活性。从瞬时表达 CYP2D6 的本塞姆塞米亚纳植物中分离出一种新型吲哚生物碱 11-羟基棒菊碱，并补充了棒菊碱，并阐明了其结构。所提出的技术可能对于实现组合生物合成概念具有价值，该组合生物合成概念包括不同代谢环境中异源酶和底物的连接。

"点击查看英文标题和摘要"

Transient Expression of Human Cytochrome P450s 2D6 and 3A4 in Nicotiana benthamiana Provides a Possibility for Rapid Substrate Testing and Production of Novel Compounds

Employment of transient expression of foreign genes for bioconversion of pharmaceutically valuable low‐molecular‐weight compounds, including plant secondary metabolites, is an enticing trend still scantily explored in plant biotechnology. In the present work, an efficient protocol for rapid assessment of synthetic and plant‐derived metabolites as potential substrates for human P450s (CYP2D6 and CYP3A4) via Agrobacterium‐mediated transient expression in Nicotiana benthamiana is put forth. Animal P450s with broad substrate specificity are promising candidates for transformation of diverse metabolites. The efficiency of P450s in heterologous surroundings is not always satisfactory and depends on the availability of an associated electron‐transfer enzyme. Plants represent an attractive assortment of prospective hosts for foreign P450s expression. The optimal composition of genetic blocks providing the highest transient expression efficiency is designed, an effective substrate administration scheme is validated, and biological activity of the investigated P450s against loratadine and several indole alkaloids with different molecular scaffold structures is tested. A novel indole alkaloid, 11‐hydroxycorynanthine, is isolated from N. benthamiana plants transiently expressing CYP2D6 and supplemented with corynanthine, and its structure was elucidated. The proposed technique might be of value in realization of combinatorial biosynthesis concept comprising the junction of heterologous enzymes and substrates in different metabolic surroundings.

更新日期：2018-09-17

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南