Reconstitution of monoterpene indole alkaloid biosynthesis in genome engineered Nicotiana benthamiana
Reconstitution of monoterpene indole alkaloid biosynthesis in genome engineered Nicotiana benthamiana
复制标题
基因组工程烟草中单萜吲哚生物碱生物合成的重建
DOI：
10.1101/2021.08.12.456143
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Dudley Q
中科院分区：
文献类型：
--
作者：
Dudley Q
单萜吲哚生物碱 (MIA) 是一类多样化的植物天然产物，其中包括许多重要的药用化合物。我们着手重建胡豆素的途径，胡豆素是所有 MIA 的关键中间体，来自本塞姆氏烟草的中央代谢。该宿主的缺点是其丰富的背景代谢导致一些异源产生的分子衍生化。在这里，我们使用转录组分析来识别响应生物合成中间体而上调的糖基转移酶，并产生编码它们的基因具有靶向突变的植物系。早期 MIA 途径在这些细胞系中的表达产生了更有利的产品特征。成功地重建了马球糖苷生物合成，通过 14 种酶的共表达获得了最佳产量，其中来自荆芥（猫薄荷）的主要乳胶蛋白样酶 (MLPL) 对于提高通过环烯醚萜途径的通量至关重要。内源糖基转移酶的去除不会影响胡胡豆素的产量，这凸显了途径酶向稳定的生物合成中间体的代谢通量最大限度地减少了改造宿主内源代谢的需要。植物胡豆素的生产扩大了适合生物合成的 MIA 产品范围。
Monoterpene indole alkaloids (MIAs) are a diverse class of plant natural products that include a number of medicinally important compounds. We set out to reconstitute the pathway for strictosidine, a key intermediate of all MIAs, from central metabolism inNicotiana benthamiana. A disadvantage of this host is that its rich background metabolism results in the derivatization of some heterologously produced molecules. Here we use transcriptomic analysis to identify glycosyltransferases that are upregulated in response to biosynthetic intermediates and produce plant lines with targeted mutations in the genes encoding them. Expression of the early MIA pathway in these lines produces a more favorable product profile. Strictosidine biosynthesis was successfully reconstituted, with the best yields obtained by the co-expression of 14 enzymes, of which a major latex protein-like enzyme (MLPL) from Nepeta (catmint) is critical for improving flux through the iridoid pathway. The removal of endogenous glycosyltransferases does not impact the yields of strictosidine, highlighting that the metabolic flux of the pathway enzymes to a stable biosynthetic intermediate minimizes the need to engineer the endogenous metabolism of the host. The production of strictosidinein plantaexpands the range of MIA products amenable to biological synthesis.
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影响因子：
3.5
作者：
Wang Q;Xing S;Pan Q;Yuan F;Zhao J;Tian Y;Chen Y;Wang G;Tang K
通讯作者：
Tang K
影响因子：
3.7
作者：
Naim F;Nakasugi K;Crowhurst RN;Hilario E;Zwart AB;Hellens RP;Taylor JM;Waterhouse PM;Wood CC
通讯作者：
Wood CC
影响因子：
15
作者：
Ishikawa H;Colby DA;Seto S;Va P;Tam A;Kakei H;Rayl TJ;Hwang I;Boger DL
通讯作者：
Boger DL
影响因子：
11.6
作者：
Murata, Jun;Roepke, Jonathon;De Luca, Vincenzo
通讯作者：
De Luca, Vincenzo
DOI：
10.1073/pnas.0403709101
发表时间：
2004-07-06
影响因子：
11.1
作者：
Song, CP;Guo, Y;Zhu, JK
通讯作者：
Zhu, JK