Large-scale robot-assisted genome shuffling yields industrial Saccharomyces cerevisiae yeasts with increased ethanol tolerance.
Large-scale robot-assisted genome shuffling yields industrial Saccharomyces cerevisiae yeasts with increased ethanol tolerance.
复制标题
大规模机器人辅助基因组改组产生的酿酒酵母酵母具有乙醇耐受性的增加。
DOI：
10.1186/s13068-015-0216-0
复制
发表时间：
2015
影响因子：
6.3
通讯作者：
Verstrepen KJ
中科院分区：
文献类型：
--
作者：
Snoek T;Picca Nicolino M;Van den Bremt S;Mertens S;Saels V;Verplaetse A;Steensels J;Verstrepen KJ
在生物乙醇发酵的最后阶段，酵母细胞面临高乙醇浓度。这种压力导致发酵减慢或停滞，并限制乙醇生产。因此，具有上级乙醇耐受性的新型酿酒酵母菌株可以允许增加的产量和效率。基因组改组已成为快速增强包括乙醇耐受性在内的复杂性状的有力方法，但以前的努力主要依赖于单一菌株的诱变池，这可能会限制有效性。在这里，我们探索新的机器人辅助策略，允许以前所未有的规模改组多个亲本酵母的基因组。对318种不同酵母的乙醇积累、孢子形成效率和遗传相关性进行筛选，产生了8种异宗配合菌株，作为基因组改组的亲本。在第一种方法中，用不同的选择方法对亲本菌株进行多轮连续的随机基因组改组，产生几种显示出增加的乙醇耐受性的杂交种。有趣的是，平均而言，第一代（F1）的杂交种比第三代（F3）的杂交种显示出更高的乙醇产量。在第二种方法中，我们应用了几轮连续的机器人辅助靶向基因组改组，产生了3,000多个靶向杂交。选择耐乙醇的杂交种表现出增加的耐乙醇性和生产相比，双杂交种，和F1杂种平均上级F3杂种。总共有135个单独的F1和F3杂交种在小规模极高浓度发酵中进行了测试。八个杂交种表现出优于商业生物燃料菌株Ethanol Red的上级发酵性能，显示出最大乙醇积累增加2至7%。在8升的中试规模中，表现最好的混合发酵培养基含有32%（w/v）的葡萄糖至干，产乙醇18.7%（v/v），生产率为0.90 g乙醇/l/h，产乙醇0.45 g乙醇/g葡萄糖。我们报告使用几种不同的大规模基因组改组策略，以获得新的杂交种增加乙醇耐受性和发酵能力。几种新的杂交种显示出最佳亲本杂种优势，并优于常用的生物乙醇菌株Ethanol Red，使它们成为工业生产的有趣候选菌株。本文的在线版本（doi：10.1186/s13068-015-0216-0）包含补充材料，可供授权用户使用。
During the final phases of bioethanol fermentation, yeast cells face high ethanol concentrations. This stress results in slower or arrested fermentations and limits ethanol production. Novel Saccharomyces cerevisiae strains with superior ethanol tolerance may therefore allow increased yield and efficiency. Genome shuffling has emerged as a powerful approach to rapidly enhance complex traits including ethanol tolerance, yet previous efforts have mostly relied on a mutagenized pool of a single strain, which can potentially limit the effectiveness. Here, we explore novel robot-assisted strategies that allow to shuffle the genomes of multiple parental yeasts on an unprecedented scale. Screening of 318 different yeasts for ethanol accumulation, sporulation efficiency, and genetic relatedness yielded eight heterothallic strains that served as parents for genome shuffling. In a first approach, the parental strains were subjected to multiple consecutive rounds of random genome shuffling with different selection methods, yielding several hybrids that showed increased ethanol tolerance. Interestingly, on average, hybrids from the first generation (F1) showed higher ethanol production than hybrids from the third generation (F3). In a second approach, we applied several successive rounds of robot-assisted targeted genome shuffling, yielding more than 3,000 targeted crosses. Hybrids selected for ethanol tolerance showed increased ethanol tolerance and production as compared to unselected hybrids, and F1 hybrids were on average superior to F3 hybrids. In total, 135 individual F1 and F3 hybrids were tested in small-scale very high gravity fermentations. Eight hybrids demonstrated superior fermentation performance over the commercial biofuel strain Ethanol Red, showing a 2 to 7% increase in maximal ethanol accumulation. In an 8-l pilot-scale test, the best-performing hybrid fermented medium containing 32% (w/v) glucose to dryness, yielding 18.7% (v/v) ethanol with a productivity of 0.90 g ethanol/l/h and a yield of 0.45 g ethanol/g glucose. We report the use of several different large-scale genome shuffling strategies to obtain novel hybrids with increased ethanol tolerance and fermentation capacity. Several of the novel hybrids show best-parent heterosis and outperform the commonly used bioethanol strain Ethanol Red, making them interesting candidate strains for industrial production. The online version of this article (doi:10.1186/s13068-015-0216-0) contains supplementary material, which is available to authorized users.
登录
查看更多内容
芽殖酵母酿酒酵母乙醇耐受性的遗传解析
影响因子：
3.3
作者：
Hu, X. H.;Wang, M. H.;Luo, Z. W.
通讯作者：
Luo, Z. W.
影响因子：
2.6
作者：
GIETZ, RD;SCHIESTL, RH;WOODS, RA
通讯作者：
WOODS, RA
影响因子：
11.3
作者：
Steensels J;Snoek T;Meersman E;Picca Nicolino M;Voordeckers K;Verstrepen KJ
通讯作者：
Verstrepen KJ
影响因子：
4.4
作者：
Ibeas, JI;Jimenez, J
通讯作者：
Jimenez, J
DOI：
10.1534/g3.113.009381
发表时间：
2014-02-19
期刊：
G3 (Bethesda, Md.)
影响因子：
--
作者：
Plech M;de Visser JA;Korona R
通讯作者：
Korona R