Biomass Refinery Research Center (BRRC), National Institute of Advanced Industrial Science and Technology (AIST), 3-11-32 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, Japan.
Microb Cell Fact. 2014 Jan 28;13:16. doi: 10.1186/1475-2859-13-16.
There has been much research on the bioconversion of xylose found in lignocellulosic biomass to ethanol by genetically engineered Saccharomyces cerevisiae. However, the rate of ethanol production from xylose in these xylose-utilizing yeast strains is quite low compared to their glucose fermentation. In this study, two diploid xylose-utilizing S. cerevisiae strains, the industrial strain MA-R4 and the laboratory strain MA-B4, were employed to investigate the differences between anaerobic fermentation of xylose and glucose, and general differences between recombinant yeast strains, through genome-wide transcription analysis.
In MA-R4, many genes related to ergosterol biosynthesis were expressed more highly with glucose than with xylose. Additionally, these ergosterol-related genes had higher transcript levels in MA-R4 than in MA-B4 during glucose fermentation. During xylose fermentation, several genes related to central metabolic pathways that typically increase during growth on non-fermentable carbon sources were expressed at higher levels in both strains. Xylose did not fully repress the genes encoding enzymes of the tricarboxylic acid and respiratory pathways, even under anaerobic conditions. In addition, several genes involved in spore wall metabolism and the uptake of ammonium, which are closely related to the starvation response, and many stress-responsive genes mediated by Msn2/4p, as well as trehalose synthase genes, increased in expression when fermenting with xylose, irrespective of the yeast strain. We further observed that transcript levels of genes involved in xylose metabolism, membrane transport functions, and ATP synthesis were higher in MA-R4 than in MA-B4 when strains were fermented with glucose or xylose.
Our transcriptomic approach revealed the molecular events underlying the response to xylose or glucose and differences between MA-R4 and MA-B4. Xylose-utilizing S. cerevisiae strains may recognize xylose as a non-fermentable carbon source, which induces a starvation response and adaptation to oxidative stress, resulting in the increased expression of stress-response genes.
已有大量研究致力于通过基因工程酿酒酵母将木质纤维素生物质中的木糖转化为乙醇。然而,与葡萄糖发酵相比,这些木糖利用酵母菌株从木糖生产乙醇的速率相当低。在这项研究中,使用了两株二倍体木糖利用酿酒酵母菌株,工业菌株 MA-R4 和实验室菌株 MA-B4,通过全基因组转录分析来研究木糖和葡萄糖厌氧发酵之间的差异,以及重组酵母菌株之间的一般差异。
在 MA-R4 中,许多与麦角固醇生物合成相关的基因在葡萄糖存在时的表达水平高于木糖存在时。此外,在葡萄糖发酵过程中,这些与麦角固醇相关的基因在 MA-R4 中的转录水平高于 MA-B4。在木糖发酵过程中,在两种菌株中,许多与中心代谢途径相关的基因的表达水平都更高,这些基因通常在非可发酵碳源上生长时会增加。即使在厌氧条件下,木糖也不能完全抑制三羧酸循环和呼吸途径的酶编码基因的表达。此外,一些与孢子壁代谢和铵吸收有关的基因,这些基因与饥饿反应密切相关,以及许多由 Msn2/4p 介导的应激响应基因,以及海藻糖合酶基因,在以木糖发酵时表达增加,与酵母菌株无关。我们进一步观察到,当菌株以葡萄糖或木糖发酵时,参与木糖代谢、膜转运功能和 ATP 合成的基因的转录水平在 MA-R4 中高于 MA-B4。
我们的转录组学方法揭示了响应木糖或葡萄糖的分子事件以及 MA-R4 和 MA-B4 之间的差异。木糖利用酿酒酵母菌株可能将木糖识别为非可发酵碳源,这会诱导饥饿反应和适应氧化应激,导致应激响应基因的表达增加。