Transcriptome analysis of Kluyveromyces marxianus under succinic acid stress and development of robust strains,Applied Microbiology and Biotechnology

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Transcriptome analysis of Kluyveromyces marxianus under succinic acid stress and development of robust strains
Applied Microbiology and Biotechnology ( IF 3.9 ) Pub Date : 2024-04-09 , DOI: 10.1007/s00253-024-13097-3
Du-Wen Zeng ₁ , Yong-Qiang Yang ₂ , Qi Wang ₁ , Feng-Li Zhang ₁ , Mao-Dong Zhang ₁ , Sha Liao ₃ , Zhi-Qiang Liu ₂ , Ya-Chao Fan ₃ , Chen-Guang Liu ₁ , Lin Zhang ₃ , Xin-Qing Zhao ₁

Affiliation

Abstract

Kluyveromyces marxianus has become an attractive non-conventional yeast cell factory due to its advantageous properties such as high thermal tolerance and rapid growth. Succinic acid (SA) is an important platform molecule that has been applied in various industries such as food, material, cosmetics, and pharmaceuticals. SA bioproduction may be compromised by its toxicity. Besides, metabolite-responsive promoters are known to be important for dynamic control of gene transcription. Therefore, studies on global gene transcription under various SA concentrations are of great importance. Here, comparative transcriptome changes of K. marxianus exposed to various concentrations of SA were analyzed. Enrichment and analysis of gene clusters revealed repression of the tricarboxylic acid cycle and glyoxylate cycle, also activation of the glycolysis pathway and genes related to ergosterol synthesis. Based on the analyses, potential SA-responsive promoters were investigated, among which the promoter strength of IMTCP2 and KLMA_50231 increased 43.4% and 154.7% in response to 15 g/L SA. In addition, overexpression of the transcription factors Gcr1, Upc2, and Ndt80 significantly increased growth under SA stress. Our results benefit understanding SA toxicity mechanisms and the development of robust yeast for organic acid production.

Key points

• Global gene transcription of K. marxianus is changed by succinic acid (SA)

• Promoter activities of IMTCP2 and KLMA_50123 are regulated by SA

• Overexpression of Gcr1, Upc2, and Ndt80 enhanced SA tolerance

Graphical Abstract

中文翻译：

琥珀酸胁迫下马克斯克鲁维酵母的转录组分析及强健菌株的开发

抽象的

马克斯克鲁维酵母因其高耐热性和快速生长等优势特性，已成为极具吸引力的非传统酵母细胞工厂。琥珀酸（SA）是一种重要的平台分子，已应用于食品、材料、化妆品、药品等各个行业。 SA 的生物生产可能会因其毒性而受到影响。此外，已知代谢物响应启动子对于基因转录的动态控制很重要。因此，研究不同SA浓度下的全局基因转录具有重要意义。在此，分析了暴露于不同浓度 SA 的马克斯克鲁维酵母的转录组变化的比较。基因簇的富集和分析揭示了三羧酸循环和乙醛酸循环的抑制，以及糖酵解途径和与麦角甾醇合成相关的基因的激活。基于分析，研究了潜在的SA响应启动子，其中IMTCP2和KLMA_50231的启动子强度响应15g/L SA而增加了43.4%和154.7%。此外，转录因子 Gcr1、Upc2 和 Ndt80 的过度表达显着增加了 SA 胁迫下的生长。我们的结果有助于了解 SA 毒性机制和开发用于有机酸生产的强大酵母。