Manipulation of the Precursor Supply in Yeast Significantly Enhances the Accumulation of Prenylated β-Carbolines
Manipulation of the Precursor Supply in Yeast Significantly Enhances the Accumulation of Prenylated β-Carbolines
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DOI：
10.1021/acssynbio.6b00387
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发表时间：
2017-06-01
影响因子：
4.7
通讯作者：
Li, Shu-Ming
中科院分区：
文献类型：
--
作者：
Backhaus, Katja;Ludwig-Radtke, Lena;Li, Shu-Ming
色氨酸衍生物1-methyl-1，2，3，4-tetrahydro-beta-carboline-3-carboxylic酸(MTCA)存在于许多植物和食品中，包括面包酵母酿酒酵母的发酵产物。MTCA由色氨酸和乙醛通过Pictet-Spengler反应生成。在本研究中，以(1S，3S)和(1R，3S)异构体以2.2：1的比例在酿酒酵母中检测到高达9 mg/L的甲氨蝶呤。据我们所知，这是关于实验室面包师酵母培养物中存在MTCA的第一份报告。在酿酒酵母中表达了三种真菌色氨酸异丙烯基转移酶基因fgaPT2、5-dmats和7-dmats，导致在吲哚环的不同位置形成了带有异戊烯基基的MTCA衍生物。这些基因在二甲基烯丙基二磷酸和色氨酸高产菌株中的表达导致了高达400 mg/L的预烯基化MTCA的产生，其比例与未预烯基化的MTCA的比例相似。所描述的物质的结构，包括它们的立体化学，通过质谱学以及一维和二维核磁共振光谱得到了明确的阐明。本研究的结果为在酿酒酵母中大量生产设计的戊烯基化MTCA提供了一种方便的系统。此外，我们的工作可以被认为是通过只引入一个关键基因来构建更复杂的分子的一个很好的例子。
The tryptophan derivative 1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (MTCA) is present in many plants and foods including fermentation products of the baker's yeast Saccharomyces cerevisiae. MTCA is formed from tryptophan and acetaldehyde via a Pictet-Spengler reaction. In this study, up to 9 mg/L of MTCA were detected as a mixture of (1S,3S) and (1R,3S) isomers in a ratio of 2.2:1 in Saccharomyces cerevisiae cultures. To the best of our knowledge, this is the first report on the presence of MTCA in laboratory baker's yeast cultures. Expression of three fungal tryptophan prenyltransferase genes, fgaPT2, 5-dmats, and 7-dmats in S. cerevisiae resulted in the formation of MTCA derivatives with prenyl moieties at different positions of the indole ring. Expression of these genes in dimethylallyl diphosphate and tryptophan overproducing strains led to generation of up to 400 mg/L of prenylated MTCAs as mixtures of (1S,3S) and (1R,3S) diastereomers in ratios similar to that of unprenylated MTCA. The structures of the described substances including their stereochemistry were unequivocally elucidated by mass spectrometry as well as one- and two-dimensional NMR spectroscopy. The results of this study provide a convenient system for the production of high amounts of designed prenylated MTCAs in S. cerevisiae. Furthermore, our work can be considered as an excellent example for the construction of more complex molecules by introducing just one key gene.