Metabolic engineering of Saccharomyces cerevisiae for the synthesis of the wine-related antioxidant resveratrol
Metabolic engineering of Saccharomyces cerevisiae for the synthesis of the wine-related antioxidant resveratrol
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DOI：
10.1016/s1567-1356(03)00157-0
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发表时间：
2003-10-01
影响因子：
3.2
通讯作者：
Pretorius, IS
中科院分区：
文献类型：
--
作者：
Becker, JVW;Armstrong, GO;Pretorius, IS
二苯乙烯类白藜芦醇是葡萄属植物在真菌侵染、伤害或紫外线照射过程中产生的应激代谢产物。白藜芦醇是在葡萄浆果的果皮中合成的，只有微量的白藜芦醇存在于果肉中。由于发酵过程中的皮肤接触，红葡萄酒比白葡萄酒含有更高的白藜芦醇浓度。除了它的抗真菌特性，白藜芦醇还被证明具有癌症化学预防活性，并降低冠心病的风险。它起到了抗氧化剂和抗诱变剂的作用，并具有诱导特定酶代谢致癌物质的能力。这项初步研究的目的是探索开发在红葡萄酒和白葡萄酒发酵过程中都能产生白藜芦醇的葡萄酒酵母的可行性，从而增加产品的健康度。为了实现这一目标，必须在酿酒酵母中引入苯丙烷途径来产生白藜芦醇合成所需的底物之一对香豆酰辅酶A。白藜芦醇合成酶的另一种底物丙二酰辅酶A已经在酵母中发现，并参与新的脂肪酸生物合成。我们推测，通过在酿酒酵母实验室菌株中共表达来自杂交杨树的辅酶A连接酶基因(4CL216)和葡萄白藜芦醇合成酶基因(Vst1)，可以实现p-香豆酰辅酶A和白藜芦醇的生产。这种酵母具有代谢对香豆酸的能力，对香豆酸是葡萄中已经存在的一种物质。因此，这种化合物被添加到实验室培养所用的合成培养基中。获得了同时表达4CL216和vst1基因的转化子，并对白藜芦醇的生产进行了测试。在对有机提取物进行β-葡萄糖苷酶处理以去除通常与白藜芦醇结合的葡萄糖部分后，结果表明酵母转化子产生了白藜芦醇β-葡萄糖苷，即白藜芦醇。这是首次报道在异种宿主中重建生物化学途径来产生白藜芦醇。(C)2003年欧洲微生物学会联合会。爱思唯尔出版，版权所有。
The stilbene resveratrol is a stress metabolite produced by Vitis vinifera grapevines during fungal infection, wounding or UV radiation. Resveratrol is synthesised particularly in the skins of grape berries and only trace amounts are present in the fruit flesh. Red wine contains a much higher resveratrol concentration than white wine, due to skin contact during fermentation. Apart from its antifungal characteristics, resveratrol has also been shown to have cancer chemopreventive activity and to reduce the risk of coronary heart disease. It acts as an antioxidant and anti-mutagen and has the ability to induce specific enzymes that metabolise carcinogenic substances. The objective of this pilot study was to investigate the feasibility of developing wine yeasts with the ability to produce resveratrol during fermentation in both red and white wines, thereby increasing the wholesomeness of the product. To achieve this goal, the phenylpropanoid pathway in Saccharomyces cerevisiae would have to be introduced to produce p-coumaroyl-CoA, one of the substrates required for resveratrol synthesis. The other substrate for resveratrol synthase, malonyl-CoA, is already found in yeast and is involved in de novo fatty-acid biosynthesis. We hypothesised that production of p-coumaroyl-CoA and resveratrol can be achieved by co-expressing the coenzyme-A ligase-encoding gene (4CL216) from a hybrid poplar and the grapevine resveratrol synthase gene (vst1) in laboratory strains of S. cerevisiae. This yeast has the ability to metabolise p-coumaric acid, a substance already present in grape must. This compound was therefore added to the synthetic media used for the growth of laboratory cultures. Transformants expressing both the 4CL216 and vst1 genes were obtained and tested for production of resveratrol. Following beta-glucosidase treatment of organic extracts for removal of glucose moieties that are typically bound to resveratrol, the results showed that the yeast transformants had produced the resveratrol beta-glucoside, piceid. This is the first report of the reconstruction of a biochemical pathway in a heterologous host to produce resveratrol. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.