Combining inhibitor tolerance and D-xylose fermentation in industrial Saccharomyces cerevisiae for efficient lignocellulose-based bioethanol production.
Combining inhibitor tolerance and D-xylose fermentation in industrial Saccharomyces cerevisiae for efficient lignocellulose-based bioethanol production.
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DOI：
10.1186/1754-6834-6-120
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发表时间：
2013-08-26
影响因子：
6.3
通讯作者：
Thevelein JM
中科院分区：
文献类型：
--
作者：
Demeke MM;Dumortier F;Li Y;Broeckx T;Foulquié-Moreno MR;Thevelein JM
除了有效的戊糖利用之外，高抑制剂耐受性是用于利用木质纤维素生物质进行经济上可行的工业生物乙醇生产的任何生物体中所需的关键性状。尽管最近的工作已经成功地在稳健的工业酿酒酵母菌株中建立了有效的木糖发酵，但所得菌株仍然缺乏足够的抑制剂耐受性以在木质纤维素水解产物中进行有效的糖发酵。本工作的目的是在单一工业酵母菌株中将高木糖发酵活性和高抑制剂耐受性结合联合收割机。我们已经筛选了580酵母菌株的高抑制剂耐受性使用underefied酸预处理云杉水解产物，并确定了一个三倍体工业面包酵母菌株具有最高的抑制剂耐受性。从该菌株中，获得了具有更高抑制剂耐受性的可交配二倍体分离体。它与新选育的具有乙醇红遗传背景的发酵D-木糖二倍体工业菌株GS 1.11 -26杂交。从四倍体杂种中筛选出819个二倍体分离子，得到两个菌株，GSF 335和GSF 767，其结合了高抑制剂耐受性和高效木糖发酵。在平行的方法中，GS1.11-26与乙醇红的单倍体分离体的减数分裂重组和104个分离体的筛选产生了类似的抑制剂耐受二倍体菌株GSE 16。与GS1.11-26相比，3株上级菌株对云杉水解液中抑制剂的耐受性显著提高，葡萄糖消耗速率、好氧生长速率和超浓发酵最大乙醇积累能力也显著提高。在复合培养基中，3株上级菌株对D-木糖的利用率为0.36 ~ 0.67 g/g DW/h，低于GS 1.11 ~ 26的1.10 g/g DW/h。另一方面，在分批发酵未酸化的酸预处理的云杉水解液中，三个上级菌株显示出与GS 1.11 -26相当的D-木糖利用率，可能是因为它们具有更高的抑制剂耐受性。与Ethanol Red相比，它们生产的乙醇多出23%。我们成功地建设了三个上级工业基地。结合了联合收割机有效的D-木糖利用和高抑制剂耐受性的酿酒酵母菌株。由于背景菌株Ethanol Red具有成功的工业应用记录，因此三个新的上级菌株具有直接应用于工业生物乙醇生产的强大潜力。
In addition to efficient pentose utilization, high inhibitor tolerance is a key trait required in any organism used for economically viable industrial bioethanol production with lignocellulose biomass. Although recent work has succeeded in establishing efficient xylose fermentation in robust industrial Saccharomyces cerevisiae strains, the resulting strains still lacked sufficient inhibitor tolerance for efficient sugar fermentation in lignocellulose hydrolysates. The aim of the present work was to combine high xylose fermentation activity and high inhibitor tolerance in a single industrial yeast strain. We have screened 580 yeast strains for high inhibitor tolerance using undetoxified acid-pretreated spruce hydrolysate and identified a triploid industrial baker’s yeast strain as having the highest inhibitor tolerance. From this strain, a mating competent diploid segregant with even higher inhibitor tolerance was obtained. It was crossed with the recently developed D-xylose fermenting diploid industrial strain GS1.11-26, with the Ethanol Red genetic background. Screening of 819 diploid segregants from the tetraploid hybrid resulted in two strains, GSF335 and GSF767, combining high inhibitor tolerance and efficient xylose fermentation. In a parallel approach, meiotic recombination of GS1.11-26 with a haploid segregant of Ethanol Red and screening of 104 segregants resulted in a similar inhibitor tolerant diploid strain, GSE16. The three superior strains exhibited significantly improved tolerance to inhibitors in spruce hydrolysate, higher glucose consumption rates, higher aerobic growth rates and higher maximal ethanol accumulation capacity in very-high gravity fermentation, compared to GS1.11-26. In complex medium, the D-xylose utilization rate by the three superior strains ranged from 0.36 to 0.67 g/g DW/h, which was lower than that of GS1.11-26 (1.10 g/g DW/h). On the other hand, in batch fermentation of undetoxified acid-pretreated spruce hydrolysate, the three superior strains showed comparable D-xylose utilization rates as GS1.11-26, probably because of their higher inhibitor tolerance. They produced up to 23% more ethanol compared to Ethanol Red. We have successfully constructed three superior industrial S. cerevisiae strains that combine efficient D-xylose utilization with high inhibitor tolerance. Since the background strain Ethanol Red has a proven record of successful industrial application, the three new superior strains have strong potential for direct application in industrial bioethanol production.
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影响因子：
4.5
作者：
Gerstein AC;Chun HJ;Grant A;Otto SP
通讯作者：
Otto SP
影响因子：
3.3
作者：
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通讯作者：
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影响因子：
6
作者：
Kim, S;Dale, BE
通讯作者：
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影响因子：
3.2
作者：
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通讯作者：
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影响因子：
6.3
作者：
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通讯作者：
Gorwa-Grauslund MF