Synthesis of Morphinan Alkaloids in Saccharomyces cerevisiae.
Synthesis of Morphinan Alkaloids in Saccharomyces cerevisiae.
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DOI：
10.1371/journal.pone.0124459
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Martin VJ
中科院分区：
文献类型：
--
作者：
Fossati E;Narcross L;Ekins A;Falgueyret JP;Martin VJ
吗啡喃生物碱是目前用于治疗中度至重度和慢性疼痛的最有效的麻醉镇痛药。研究了从前体 (R,S)-降劳丹索林开始在重组酿酒酵母中合成吗啡喃的可行性。对罂粟甲基转移酶表达产生的网状番荔枝碱的手性分析表明，从 (R,S)-降劳丹索林开始，对 (S)-网状番荔枝碱具有严格的对映选择性。此外，P. somniferum酶salutaridinol-7-O-acetate和salutaridinol-7-O-acetate的有效自发重排和从(R)-网状番荔枝碱合成蒂巴因在酿酒酵母中功能性共表达。最后，我们重建了从 (R)-网状番荔枝碱生产可待因和吗啡的 7 基因途径。使用 (R)-网状番荔枝碱、salutaridine 或可待因作为底物的酵母细胞喂养测定表明，所有酶在酵母中均功能性共表达，并且 salutaridine 还原酶和可待因-O-脱甲基酶的活性可能限制吗啡合成的通量。这项研究的结果描述了在酿酒酵母中合成吗啡喃的重大进展，并为其在重组微生物中的完全合成铺平了道路。
Morphinan alkaloids are the most powerful narcotic analgesics currently used to treat moderate to severe and chronic pain. The feasibility of morphinan synthesis in recombinant Saccharomyces cerevisiae starting from the precursor (R,S)-norlaudanosoline was investigated. Chiral analysis of the reticuline produced by the expression of opium poppy methyltransferases showed strict enantioselectivity for (S)-reticuline starting from (R,S)-norlaudanosoline. In addition, the P. somniferum enzymes salutaridine synthase (PsSAS), salutaridine reductase (PsSAR) and salutaridinol acetyltransferase (PsSAT) were functionally co-expressed in S. cerevisiae and optimization of the pH conditions allowed for productive spontaneous rearrangement of salutaridinol-7-O-acetate and synthesis of thebaine from (R)-reticuline. Finally, we reconstituted a 7-gene pathway for the production of codeine and morphine from (R)-reticuline. Yeast cell feeding assays using (R)-reticuline, salutaridine or codeine as substrates showed that all enzymes were functionally co-expressed in yeast and that activity of salutaridine reductase and codeine-O-demethylase likely limit flux to morphine synthesis. The results of this study describe a significant advance for the synthesis of morphinans in S. cerevisiae and pave the way for their complete synthesis in recombinant microbes.
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影响因子：
16.6
作者：
通讯作者：
--
影响因子：
3.8
作者：
GERARDY, R;ZENK, MH
通讯作者：
ZENK, MH
影响因子：
2.8
作者：
Biancofiore G
通讯作者：
Biancofiore G
影响因子：
4.8
作者：
Ilari, Andrea;Franceschini, Stefano;Boffi, Alberto
通讯作者：
Boffi, Alberto
影响因子：
3.8
作者：
DEEKNAMKUL, W;ZENK, MH
通讯作者：
ZENK, MH