Improvement of Saccharomyces cerevisiae strain tolerance to vanillin through heavy ion radiation combined with adaptive laboratory evolution

IF 4.1 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of biotechnology Pub Date : 2024-08-27 DOI:10.1016/j.jbiotec.2024.08.014
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Abstract

Vanillin is an inhibitor of lignocellulose hydrolysate, which can reduce the ability of Saccharomyces cerevisiae to utilize lignocellulose, which is an important factor limiting the development of the ethanol fermentation industry. In this study, mutants of vanillin-tolerant yeast named H6, H7, X3, and X8 were bred by heavy ion irradiation (HIR) combined with adaptive laboratory evolution (ALE). Phenotypic tests revealed that the mutants outperformed the original strain WT in tolerance, growth rate, genetic stability and fermentation ability. At 1.6 g/L vanillin concentration, the average OD600 value obtained for mutant strains was 0.95 and thus about 3.4-fold higher than for the wild-type. When the concentration of vanillin was 2.0 g/L, the glucose utilization rate of the mutant was 86.3 % within 96 h, while that of the original strain was only 70.0 %. At this concentration of vanillin, the mitochondrial membrane potential of the mutant strain recovered faster than that of the original strain, and the ROS scavenging ability was stronger. We analyzed the whole transcriptome sequencing map and the whole genome resequencing of the mutant, and found that DEGs such as FLO9, GRC3, PSP2 and SWF1, which have large differential expression multiples and obvious mutation characteristics, play an important role in cell flocculation, rDNA transcription, inhibition of DNA polymerase mutation and protein palmitoylation. These functions can help cells resist vanillin stress. The results show that combining HIR with ALE is an effective mutagenesis strategy. This approach can efficiently obtain Saccharomyces cerevisiae mutants with improved vanillin tolerance, and provide reference for obtaining robust yeast strains with lignocellulose inhibitor tolerance.

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通过重离子辐射结合实验室适应性进化提高酿酒酵母菌株对香兰素的耐受性。
香兰素是木质纤维素水解物的抑制剂,可降低酿酒酵母利用木质纤维素的能力,是制约乙醇发酵工业发展的重要因素。本研究通过重离子辐照(HIR)结合实验室适应性进化(ALE)培育出了耐香兰素酵母突变体H6、H7、X3和X8。表型测试表明,突变体在耐受性、生长速度、遗传稳定性和发酵能力方面均优于原始菌株 WT。当香兰素浓度为 1.6 克/升时,突变株的平均 OD600 值为 0.95,因此比野生型高出约 3.4 倍。当香兰素浓度为 2.0 克/升时,突变体在 96 小时内的葡萄糖利用率为 86.3%,而原始菌株的葡萄糖利用率仅为 70.0%。在此香兰素浓度下,突变株的线粒体膜电位恢复速度比原菌株快,清除 ROS 的能力更强。我们分析了突变株的全转录组测序图谱和全基因组重测序,发现FLO9、GRC3、PSP2和SWF1等DEGs具有较大的差异表达倍数和明显的突变特征,在细胞絮凝、rDNA转录、抑制DNA聚合酶突变和蛋白质棕榈酰化等方面发挥重要作用。这些功能可以帮助细胞抵抗香兰素应激。研究结果表明,将 HIR 与 ALE 结合是一种有效的诱变策略。这种方法可以有效地获得具有更好的香兰素耐受性的酿酒酵母突变体,并为获得具有木质纤维素抑制剂耐受性的健壮酵母菌株提供参考。
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来源期刊
Journal of biotechnology
Journal of biotechnology 工程技术-生物工程与应用微生物
CiteScore
8.90
自引率
2.40%
发文量
190
审稿时长
45 days
期刊介绍: The Journal of Biotechnology has an open access mirror journal, the Journal of Biotechnology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. The Journal provides a medium for the rapid publication of both full-length articles and short communications on novel and innovative aspects of biotechnology. The Journal will accept papers ranging from genetic or molecular biological positions to those covering biochemical, chemical or bioprocess engineering aspects as well as computer application of new software concepts, provided that in each case the material is directly relevant to biotechnological systems. Papers presenting information of a multidisciplinary nature that would not be suitable for publication in a journal devoted to a single discipline, are particularly welcome.
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