Xiao Zhiqiang, Zhao Yifei, Wang Yongtong, Tan Xinjia, Wang Lian, Mao Jiwei, Zhang Siqi, Lu Qiyuan, Hu Fanglin, Zuo Shasha, Liu Juan, Shan Yang
Longping Agricultural College, Hunan University, Changsha, 410125, China.
Hunan Institute of Agricultural Product Processing and Quality Safety, DongTing Laboratory, Hunan Academy of Agricultural Sciences, Changsha, 410125, China.
Nat Commun. 2025 Jun 5;16(1):5211. doi: 10.1038/s41467-025-60578-8.
Saccharomyces cerevisiae primarily generates energy through glycolysis and respiration. However, the manifestation of the Crabtree effect results in substantial carbon loss and energy inefficiency, which significantly diminishes product yield and escalates substrate costs in microbial cell factories. To address this challenge, we introduce the sucrose phosphorolysis pathway and delete the phosphoglucose isomerase gene PGI1, effectively decoupling glycolysis from respiration and facilitating the metabolic transition of yeast to a Crabtree-negative state. Additionally, a synthetic energy system is engineered to regulate the NADH/NAD ratio, ensuring sufficient ATP supply and maintaining redox balance for optimal growth. The reprogrammed yeast strain exhibits significantly higher yields of various non-ethanol compounds, with lactic acid and 3-hydroxypropionic acid production increasing by 8- to 11-fold comparing to the conventional Crabtree-positive strain. This study describes an approach for overcoming the Crabtree effect in yeast, substantially improving energy metabolism, carbon recovery, and product yields.
酿酒酵母主要通过糖酵解和呼吸作用产生能量。然而,克勒勃屈利效应的表现导致大量碳损失和能量利用效率低下,这显著降低了微生物细胞工厂中的产品产量并提高了底物成本。为应对这一挑战,我们引入了蔗糖磷酸解途径并删除了磷酸葡萄糖异构酶基因PGI1,有效地使糖酵解与呼吸作用脱钩,并促进酵母向克勒勃屈利阴性状态的代谢转变。此外,设计了一种合成能量系统来调节NADH/NAD比率,确保充足的ATP供应并维持氧化还原平衡以实现最佳生长。重新编程的酵母菌株表现出各种非乙醇化合物的产量显著提高,与传统的克勒勃屈利阳性菌株相比,乳酸和3-羟基丙酸的产量增加了8至11倍。本研究描述了一种克服酵母中克勒勃屈利效应的方法,可大幅改善能量代谢、碳回收和产品产量。
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