Transcriptome analysis and reverse engineering verification of SNZ3Val125Ile and Pho3Asn134Asp revealed the mechanism of adaptive laboratory evolution to increase the yield of tyrosol in Saccharomyces cerevisiae strain S26-AE2

IF 6.1 1区工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology for Biofuels Pub Date : 2025-03-05 DOI:10.1186/s13068-025-02627-4

Na Song, Huili Xia, Xiaoxue Yang, Siyao Liu, Linglong Xu, Kun Zhuang, Lan Yao, Shihui Yang, Xiong Chen, Jun Dai

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Abstract

Background

Tyrosol is an important drug precursor, and Saccharomyces cerevisiae is one of the main microorganisms that produces tyrosol. Although excessive metabolic modification increases the production of tyrosol, it also causes a decrease in the growth rate of yeast. Therefore, this study attempted to restore the growth of S. cerevisiae through adaptive evolution and further improve tyrosol production.

Results

After the adaptive laboratory evolution of S. cerevisiae S26, three evolutionary strains were obtained. The biomass of strain S26-AE2 reached 17.82 g DCW/L in the presence of 100 g/L glucose, which was 15.33% higher than that of S26, and its tyrosol production reached 817.83 mg/L. The transcriptome analysis revealed that, upon exposure to 100 g/L glucose, the S26-AE2 strain may reduce the transcriptional regulation of glucose repression through decreased HXK2 expression. The expression of genes related to pyruvate synthesis was increased in strain S26-AE2. Meanwhile, the expression levels of most tricarboxylic acid cycle-related genes in S26-AE2 were increased when cultured with 20 g/L glucose. Furthermore, the amount of tyrosol produced by strain S26 with the SNZ3^Val125Ile mutation increased by 17.01% compared with that of the control strain S26 following exposure to 100 g/L glucose.

Conclusions

In this study, a strain, S26-AE2, with good growth and tyrosol production performance was obtained by adaptive evolution. The transcriptome analysis revealed that the differences in the expression of genes involved in metabolic pathways in adaptive evolutionary strains may be related to yeast growth and tyrosol production. Further reverse engineering verified that the mutation of SNZ3 promoted tyrosol synthesis in S. cerevisiae in glucose-rich medium. This study provides a theoretical basis for the metabolic engineering of S. cerevisiae to synthesise tyrosol and its derivatives.

Graphical Abstract

查看原文本刊更多论文

通过对SNZ3Val125Ile和Pho3Asn134Asp的转录组分析和逆向工程验证，揭示了酿酒酵母菌S26-AE2的适应性实验室进化提高酪醇产量的机制

背景酪醇是重要的药物前体，酿酒酵母是产生酪醇的主要微生物之一。虽然过度的代谢修饰增加了酪醇的产量，但它也会导致酵母生长速度的下降。因此，本研究试图通过适应性进化恢复酿酒酵母的生长，进一步提高酪醇的产量。结果酿酒葡萄球菌S26经过实验室适应性进化，获得3个进化菌株。菌株S26- ae2在100 g/L葡萄糖条件下的生物量达到17.82 g DCW/L，比菌株S26提高了15.33%，其酪醇产量达到817.83 mg/L。转录组分析显示，当暴露于100 g/L葡萄糖时，S26-AE2菌株可能通过降低HXK2表达来降低葡萄糖抑制的转录调节。菌株S26-AE2中丙酮酸合成相关基因的表达增加。与此同时，在20 g/L葡萄糖的培养下，S26-AE2中大部分三羧酸循环相关基因的表达量均增加。此外，SNZ3Val125Ile突变菌株S26与对照菌株S26相比，在暴露于100 g/L葡萄糖后，酪醇的产量增加了17.01%。结论本研究通过自适应进化获得了一株生长和产酪醇性能良好的菌株S26-AE2。转录组分析显示，适应性进化菌株中代谢途径相关基因的表达差异可能与酵母生长和酪醇生产有关。进一步的逆向工程证实SNZ3突变促进了酿酒酵母在富葡萄糖培养基中酪醇的合成。本研究为酿酒酵母代谢工程合成酪醇及其衍生物提供了理论依据。图形抽象

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来源期刊

Biotechnology for Biofuels 工程技术-生物工程与应用微生物

自引率

0.00%

发文量

审稿时长

2.7 months

期刊介绍： Biotechnology for Biofuels is an open access peer-reviewed journal featuring high-quality studies describing technological and operational advances in the production of biofuels, chemicals and other bioproducts. The journal emphasizes understanding and advancing the application of biotechnology and synergistic operations to improve plants and biological conversion systems for the biological production of these products from biomass, intermediates derived from biomass, or CO2, as well as upstream or downstream operations that are integral to biological conversion of biomass. Biotechnology for Biofuels focuses on the following areas: • Development of terrestrial plant feedstocks • Development of algal feedstocks • Biomass pretreatment, fractionation and extraction for biological conversion • Enzyme engineering, production and analysis • Bacterial genetics, physiology and metabolic engineering • Fungal/yeast genetics, physiology and metabolic engineering • Fermentation, biocatalytic conversion and reaction dynamics • Biological production of chemicals and bioproducts from biomass • Anaerobic digestion, biohydrogen and bioelectricity • Bioprocess integration, techno-economic analysis, modelling and policy • Life cycle assessment and environmental impact analysis