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首页> 外文期刊>Applied Microbiology >Himalayan Saccharomyces eubayanus Genome Sequences Reveal Genetic Markers Explaining Heterotic Maltotriose Consumption by Saccharomyces pastorianus Hybrids
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Himalayan Saccharomyces eubayanus Genome Sequences Reveal Genetic Markers Explaining Heterotic Maltotriose Consumption by Saccharomyces pastorianus Hybrids

机译：喜马拉雅酿酒酵母eubayanus基因组序列揭示了遗传标记，解释了酵母菌对杂种麦芽三糖的消费。

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Saccharomyces pastorianus strains are hybrids of Saccharomyces cerevisiae and Saccharomyces eubayanus that have been domesticated for centuries in lager beer brewing environments. As sequences and structures of S. pastorianus genomes are being resolved, molecular mechanisms and evolutionary origins of several industrially relevant phenotypes remain unknown. This study investigates how maltotriose metabolism, a key feature in brewing, may have arisen in early S. eubayanus × S. cerevisiae hybrids. To address this question, we generated a nearly complete genome assembly of Himalayan S. eubayanus strains of the Holarctic subclade. This group of strains has been proposed to be the S. eubayanus subgenome origin of current S. pastorianus strains. The Himalayan S. eubayanus genomes harbored several copies of an S. eubayanusAGT1 (SeAGT1) α-oligoglucoside transporter gene with high sequence identity to genes encountered in S. pastorianus. Although Himalayan S. eubayanus strains cannot grow on maltose and maltotriose, their maltose-hydrolase and SeMALT1 and SeAGT1 maltose transporter genes complemented the corresponding null mutants of S. cerevisiae. Expression, in Himalayan S. eubayanus of a functional S. cerevisiae maltose metabolism regulator gene (MALx3) enabled growth on oligoglucosides. The hypothesis that the maltotriose-positive phenotype in S. pastorianus is a result of heterosis was experimentally tested by constructing an S. cerevisiae × S. eubayanus laboratory hybrid with a complement of maltose metabolism genes that resembles that of current S. pastorianus strains. The ability of this hybrid to consume maltotriose in brewer’s wort demonstrated regulatory cross talk between subgenomes and thereby validated this hypothesis. These results support experimentally the new postulated hypothesis on the evolutionary origin of an essential phenotype of lager brewing strains and valuable knowledge for industrial exploitation of laboratory-made S. pastorianus-like hybrids.IMPORTANCES. pastorianus, an S. cerevisiae × S. eubayanus hybrid, is used for production of lager beer, the most produced alcoholic beverage worldwide. It emerged by spontaneous hybridization and colonized early lager brewing processes. Despite accumulation and analysis of genome sequencing data of S. pastorianus parental genomes, the genetic blueprint of industrially relevant phenotypes remains unresolved. Assimilation of maltotriose, an abundant sugar in wort, has been postulated to be inherited from the S. cerevisiae parent. Here, we demonstrate that although Asian S. eubayanus isolates harbor a functional maltotriose transporter SeAGT1 gene, they are unable to grow on α-oligoglucosides, but expression of S. cerevisiae regulator MAL13 (ScMAL13) was sufficient to restore growth on trisaccharides. We hypothesized that the S. pastorianus maltotriose phenotype results from regulatory interaction between S. cerevisiae maltose transcription activator and the promoter of SeAGT1. We experimentally confirmed the heterotic nature of the phenotype, and thus these results provide experimental evidence of the evolutionary origin of an essential phenotype of lager brewing strains.
机译：巴斯德酿酒酵母菌株是酿酒酵母和欧亚酿酒酵母的杂种，已经在大型啤酒酿造环境中驯化了几个世纪。在解决巴氏链球菌基因组的序列和结构时，几种与工业相关的表型的分子机制和进化起源仍然未知。这项研究调查了早期酿酒酵母×酿酒酵母杂交物中麦芽三糖代谢（酿造的关键特征）如何发生。为了解决这个问题，我们生成了Holarctic子群喜马拉雅S. eubayanus菌株的几乎完整的基因组组装体。已经提出这组菌株是当前巴氏巴氏酵母菌株的欧亚巴氏酵母亚基因组起源。喜马拉雅葡萄球菌的基因组具有几个拷贝的葡萄球菌AGT1（SeAGT1）α-寡糖苷转运蛋白基因，该基因与巴斯德酵母中遇到的基因具有高度序列同一性。尽管喜马拉雅葡萄球菌不能在麦芽糖和麦芽三糖上生长，但它们的麦芽糖水解酶以及SeMALT1和SeAGT1麦芽糖转运蛋白基因补充了酿酒酵母的相应无效突变体。功能性酿酒酵母麦芽糖代谢调节基因（MALx3）在喜马拉雅芥中的表达使寡糖苷能够生长。通过构建具有与目前的巴氏酵母菌株相似的麦芽糖代谢基因的互补序列的酿酒酵母×欧巴氏酵母实验室杂交体，通过实验检验了巴氏酵母中麦芽三糖阳性表型是杂种优势的假说。这种杂种在啤酒麦芽汁中消耗麦芽三糖的能力证明了亚基因组之间的调节性串扰，从而验证了这一假设。这些结果在实验上支持了关于较大啤酒酿造菌株基本表型的进化起源的新假设假说，以及对实验室制造的巴氏链球菌样杂种的工业开发的宝贵知识。 pastorianus是一种酿酒酵母×欧亚酵母（S. eubayanus）杂种，用于生产较大的啤酒，这是世界上产量最高的酒精饮料。它通过自发杂交和定殖的早期啤酒酿造过程出现。尽管积累和分析了巴氏链球菌亲本基因组的基因组测序数据，但仍未解析工业上相关表型的遗传蓝图。麦芽三糖（麦芽汁中的大量糖）的同化被认为是从酿酒酵母的亲本遗传而来的。在这里，我们证明，尽管亚洲葡萄球菌菌株具有功能性的麦芽三糖转运蛋白SeAGT1基因，但它们无法在α-寡糖苷上生长，但是酿酒酵母调节物MAL13（ScMAL13）的表达足以恢复三糖的生长。我们假设S. pastorianus麦芽三糖表型是由酿酒酵母麦芽糖转录激活因子和SeAGT1启动子之间的调控相互作用引起的。我们通过实验证实了表型的异质性，因此这些结果提供了啤酒酿造菌株基本表型进化起源的实验证据。

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