Enhanced acetic acid stress tolerance and ethanol production in Saccharomyces cerevisiae by modulating expression of the de novo purine biosynthesis genes
Enhanced acetic acid stress tolerance and ethanol production in Saccharomyces cerevisiae by modulating expression of the de novo purine biosynthesis genes
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通过调节从头嘌呤生物合成基因的表达增强酿酒酵母的乙酸胁迫耐受性和乙醇产量
DOI：
10.1186/s13068-019-1456-1
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发表时间：
2019-05
影响因子：
6.3
通讯作者：
Zhao Xin-qing
中科院分区：
文献类型：
--
作者：
Zhang Ming Ming;Xiong Liang;Tang Ya Jie;Mehmood M. Aamer;Zhao Zongbao Kent;Bai Feng-Wu;Zhao Xin-qing
背景耐受多种环境胁迫的YEast菌株在各种工业应用中都是非常需要的。尽管人们在寻找与酿酒酵母耐逆相关的关键基因方面做了大量的工作，但新的嘌呤生物合成基因在酵母耐逆中的作用还没有得到很好的研究。我们之前的研究表明，添加硫酸锌提高了酵母对醋酸的耐受性，并对参与酵母耐受胁迫的关键基因进行了进一步的研究。结果在醋酸胁迫下，添加硫酸锌显著提高了3个参与从头合成嘌呤生物合成的基因ADE1、ADE13和ADE17的转录水平，这些基因在酿酒酵母BY4741中的过表达促进了不同胁迫条件下的细胞生长。同时，3个ADE基因在胁迫条件下的过表达也提高了乙醇产量，其中在木质纤维生物质混合抑制剂存在下，ADE17的过表达提高幅度最高，达到158.39%。结果表明，无论在对照条件下还是在醋酸胁迫条件下，ADE17过表达均能提高腺嘌呤核苷酸库 + 和 + 含量，这与重组酵母较好的生长状态是一致的。ADE基因的过度表达也改变了全球细胞内氨基酸组成。在这些变化的氨基酸中，由于ade基因在醋酸胁迫下的过表达，导致胁迫保护剂γ-氨基丁酸的显著增加，这表明ade基因的过表达对嘌呤生物合成和氨基酸生物合成都起到控制作用，从而保护酵母细胞免受胁迫。结论我们证明从头合成嘌呤生物合成基因是酵母耐逆代谢工程的有用靶点。本研究开发的工程菌株对多种抑制剂的耐受性提高，可用于高效的木质纤维生物精炼生产生物燃料和生物化学品。
BackgroundYeast strains that are tolerant to multiple environmental stresses are highly desired for various industrial applications. Despite great efforts in identifying key genes involved in stress tolerance of budding yeast Saccharomyces cerevisiae, the effects of de novo purine biosynthesis genes on yeast stress tolerance are still not well explored. Our previous studies showed that zinc sulfate addition improved yeast acetic acid tolerance, and key genes involved in yeast stress tolerance were further investigated in this study.ResultsThree genes involved in de novo purine biosynthesis, namely, ADE1, ADE13, and ADE17, showed significantly increased transcription levels by zinc sulfate supplementation under acetic acid stress, and overexpression of these genes in S. cerevisiae BY4741 enhanced cell growth under various stress conditions. Meanwhile, ethanol productivity was also improved by overexpression of the three ADE genes under stress conditions, among which the highest improvement attained 158.39% by ADE17 overexpression in the presence of inhibitor mixtures derived from lignocellulosic biomass. Elevated levels of adenine-nucleotide pool “AXP” ([ATP] + [ADP] + [AMP]) and ATP content were observed by overexpression of ADE17, both under control condition and under acetic acid stress, and is consistent with the better growth of the recombinant yeast strain. The global intracellular amino acid profiles were also changed by overexpression of the ADE genes. Among the changed amino acids, significant increase of the stress protectant γ-aminobutyric acid (GABA) was revealed by overexpression of the ADE genes under acetic acid stress, suggesting that overexpression of the ADE genes exerts control on both purine biosynthesis and amino acid biosynthesis to protect yeast cells against the stress.ConclusionWe proved that the de novo purine biosynthesis genes are useful targets for metabolic engineering of yeast stress tolerance. The engineered strains developed in this study with improved tolerance against multiple inhibitors can be employed for efficient lignocellulosic biorefinery to produce biofuels and biochemicals.
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影响因子：
3.6
作者：
Pagani, M. Ayelen;Casamayor, Antonio;Arino, Joaquin
通讯作者：
Arino, Joaquin
影响因子：
5.3
作者：
通讯作者：
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影响因子：
6.4
作者：
Mira NP;Palma M;Guerreiro JF;Sá-Correia I
通讯作者：
Sá-Correia I
影响因子：
6.3
作者：
González-Ramos D;Gorter de Vries AR;Grijseels SS;van Berkum MC;Swinnen S;van den Broek M;Nevoigt E;Daran JM;Pronk JT;van Maris AJ
通讯作者：
van Maris AJ
影响因子：
6.3
作者：
Chen Y;Sheng J;Jiang T;Stevens J;Feng X;Wei N
通讯作者：
Wei N