De novo biosynthesis of diverse plant-derived styrylpyrones in Saccharomyces cerevisiae.
De novo biosynthesis of diverse plant-derived styrylpyrones in Saccharomyces cerevisiae.
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DOI：
10.1016/j.mec.2022.e00195
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发表时间：
2022-06
影响因子：
5.2
通讯作者：
Li S
中科院分区：
文献类型：
--
作者：
Wu Y;Chen MN;Li S
近年来，植物苯乙烯基吡喃酮类化合物发挥了良好的神经保护作用，引起了人们越来越多的关注。在工程微生物中合成每一个苯乙烯基吡喃酮的能力对于理解药用植物的生物活性及其产生的复杂混合物是重要的。不同植物来源的苯乙烯基吡喃酮的微生物生物制造还提供了用于生产有价值的植物苯乙烯基吡喃酮的可持续和有效的方法，作为对流行的基于农业的方法的补充的日常补充剂或潜在药物。在本研究中，我们首次证实了两种7，8-饱和苯乙烯基吡喃酮（7，8-dihydro-5，6-dehydrokavain（DDK）和7，8-dihydroyangonin（DHY））和两种7，8-不饱和苯乙烯基吡喃酮（desmethoxyyangonin（DMY）和yangonin（Y））在酿酒酵母中的异源生物合成。虽然植物苯乙烯基吡喃酮生物合成途径尚未完全阐明，我们功能性地重建了最近发现的卡瓦苯乙烯基吡喃酮生物合成途径，在酵母中具有高底物混杂性，并将其与上游羟基肉桂酸生物合成途径相结合，以产生多种植物来源的苯乙烯基吡喃酮，而无需天然植物酶。我们通过工程改造酵母内源性芳香族氨基酸代谢和内源性双键还原酶以及通过CRISPR介导的δ整合来过表达限速途径基因来优化从头途径。这些组合工程的努力导致了最初的三种酵母菌株，它们可以从头产生不同的植物来源的苯乙烯基吡喃酮，DDK，DMY和Y的滴度分别为4.40 μM，1.28 μM和0.10 μM。本研究为苯乙烯吡喃酮的大规模生物制备和更复杂植物苯乙烯吡喃酮的完全生物合成奠定了基础。植物苯乙烯基吡喃酮的完全生物合成首先在酵母中实现。酵母酶代替未知的植物酶产生7，8-饱和苯乙烯基吡喃酮。基于CRISPR的δ-整合导致在丰富培养基中稳定的苯乙烯基吡喃酮过量生产。
Plant styrylpyrones exerting well-established neuroprotective properties have attracted increasing attention in recent years. The ability to synthesize each individual styrylpyrone in engineered microorganisms is important to understanding the biological activity of medicinal plants and the complex mixtures they produce. Microbial biomanufacturing of diverse plant-derived styrylpyrones also provides a sustainable and efficient approach for the production of valuable plant styrylpyrones as daily supplements or potential drugs complementary to the prevalent agriculture-based approach. In this study, we firstly demonstrated the heterogenous biosynthesis of two 7,8-saturated styrylpyrones (7,8-dihydro-5,6-dehydrokavain (DDK) and 7,8-dihydroyangonin (DHY)) and two 7,8-unsaturated styrylpyrones (desmethoxyyangonin (DMY) and yangonin (Y)), in Saccharomyces cerevisiae. Although plant styrylpyrone biosynthetic pathways have not been fully elucidated, we functionally reconstructed the recently discovered kava styrylpyrone biosynthetic pathway that has high substrate promiscuity in yeast, and combined it with upstream hydroxycinnamic acid biosynthetic pathways to produce diverse plant-derived styrylpyrones without the native plant enzymes. We optimized the de novo pathways by engineering yeast endogenous aromatic amino acid metabolism and endogenous double bond reductases and by CRISPR-mediated δ-integration to overexpress the rate-limiting pathway genes. These combinatorial engineering efforts led to the first three yeast strains that can produce diverse plant-derived styrylpyrones de novo, with the titers of DDK, DMY and Y at 4.40 μM, 1.28 μM and 0.10 μM, respectively. This work has laid the foundation for larger-scale styrylpyrone biomanufacturing and the complete biosynthesis of more complicated plant styrylpyrones. Complete biosynthesis of plant styrylpyrones was firstly achieved in yeast. Yeast enzyme replaces unknown plant enzymes to produce 7,8-saturated styrylpyrones. CRISPR-based δ-integration led to stable styrylpyrone overproduction in rich medium.
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影响因子：
6.4
作者：
Gold ND;Gowen CM;Lussier FX;Cautha SC;Mahadevan R;Martin VJ
通讯作者：
Martin VJ
影响因子：
4.4
作者：
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5.2
作者：
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通讯作者：
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影响因子：
48
作者：
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通讯作者：
Smith, Hamilton O.
影响因子：
3.2
作者：
Lehka BJ;Eichenberger M;Bjørn-Yoshimoto WE;Vanegas KG;Buijs N;Jensen NB;Dyekjær JD;Jenssen H;Simon E;Naesby M
通讯作者：
Naesby M