Regulation of the PFK1 gene on the interspecies microbial competition behavior of Saccharomyces cerevisiae,Applied Microbiology and Biotechnology

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Regulation of the PFK1 gene on the interspecies microbial competition behavior of Saccharomyces cerevisiae
Applied Microbiology and Biotechnology ( IF 3.9 ) Pub Date : 2024-03-22 , DOI: 10.1007/s00253-024-13091-9
Caijuan Zheng ₁ , Shuxin Hou ₁ , Yu Zhou ₂ , Changyuan Yu ₁ , Hao Li ₂

Affiliation

Abstract

Saccharomyces cerevisiae is a widely used strain for ethanol fermentation; meanwhile, efficient utilization of glucose could effectively promote ethanol production. The PFK1 gene is a key gene for intracellular glucose metabolism in S. cerevisiae. Our previous work suggested that although deletion of the PFK1 gene could confer higher oxidative tolerance to S. cerevisiae cells, the PFK1Δ strain was prone to contamination by other microorganisms. High interspecies microbial competition ability is vital for the growth and survival of microorganisms in co-cultures. The result of our previous studies hinted us a reasonable logic that the EMP (i.e., the Embden-Meyerhof-Parnas pathway, the glycolytic pathway) key gene PFK1 could be involved in regulating interspecies competitiveness of S. cerevisiae through the regulation of glucose utilization and ethanol production efficiency. The results suggest that under 2% and 5% glucose, the PFK1Δ strain showed slower growth than the S288c wild-type and TDH1Δ strains in the lag and exponential growth stages, but realized higher growth in the stationary stage. However, relative high supplement of glucose (10%) eliminated this phenomenon, suggesting the importance of glucose in the regulation of PFK1 in yeast cell growth. Furthermore, during the lag growth phase, the PFK1Δ strain displayed a decelerated glucose consumption rate (P < 0.05). The expression levels of the HXT2, HXT5, and HXT6 genes decreased by approximately 0.5-fold (P < 0.05) and the expression level of the ZWF1 exhibited a onefold increase in the PFK1Δ strain compared to that in the S. cerevisiae S288c wild-type strain (P < 0.05).These findings suggested that the PFK1 inhibited the uptake and utilization of intracellular glucose by yeast cells, resulting in a higher amount of residual glucose in the medium for the PFK1Δ strain to utilize for growth during the reverse overshoot stage in the stationary phase. The results presented here also indicated the potential of ethanol as a defensive weapon against S. cerevisiae. The lower ethanol yield in the early stage of the PFK1Δ strain (P < 0.001) and the decreased expression levels of the PDC5 and PDC6 (P < 0.05), which led to slower growth, resulted in the strain being less competitive than the wild-type strain when co-cultured with Escherichia coli. The lower interspecies competitiveness of the PFK1Δ strain further promoted the growth of co-cultured E. coli, which in turn activated the ethanol production efficiency of the PFK1Δ strain to antagonize it from E. coli at the stationary stage. The results presented clarified the regulation of the PFK1 gene on the growth and interspecies microbial competition behavior of S. cerevisiae and would help us to understand the microbial interactions between S. cerevisiae and other microorganisms.

Key points

• PFK1Δ strain could realize reverse growth overshoot at the stationary stage

• PFK1 deletion decreased ethanol yield and interspecific competitiveness

• Proportion of E. coli in co-culture affected ethanol yield capacity of yeast cells

中文翻译：

PFK1基因对酿酒酵母种间微生物竞争行为的调控

抽象的

酿酒酵母是一种广泛用于乙醇发酵的菌株；同时，葡萄糖的高效利用可以有效促进乙醇生产。 PFK1基因是酿酒酵母细胞内葡萄糖代谢的关键基因。我们之前的工作表明，虽然PFK1基因的缺失可以赋予酿酒酵母细胞更高的氧化耐受性，但PFK1Δ菌株很容易受到其他微生物的污染。微生物的高种间竞争能力对于共培养中微生物的生长和生存至关重要。我们前期的研究结果给我们提示了一个合理的逻辑：EMP（即Embden-Meyerhof-Parnas途径，糖酵解途径）关键基因PFK1可能通过调节葡萄糖的利用和利用来参与调节酿酒酵母的种间竞争力。乙醇生产效率。结果表明，在2%和5%葡萄糖下， PFK1Δ菌株在滞后期和指数生长期表现出比S288c野生型和TDH1Δ菌株更慢的生长，但在稳定期实现了更高的生长。然而，相对较高的葡萄糖补充量（10%）消除了这种现象，表明葡萄糖在酵母细胞生长中调节PFK1的重要性。此外，在生长迟缓阶段， PFK1Δ菌株表现出葡萄糖消耗率减慢（ P <0.05）。 HXT2 、 HXT5和HXT6基因的表达水平降低了约 0.5 倍（ P < 0.05），与酿酒酵母S288c野生型菌株相比， PFK1Δ菌株中ZWF1的表达水平增加了一倍（ P < 0.05）。这些结果表明PFK1抑制了细胞内葡萄糖的摄取和利用酵母细胞的作用，导致培养基中残留葡萄糖量较高，供PFK1Δ菌株在稳定期的反向过冲阶段用于生长。这里提出的结果还表明乙醇作为对抗酿酒酵母的防御武器的潜力。 PFK1Δ菌株早期乙醇产量较低（ P < 0.001）， PDC5和PDC6表达水平降低（ P < 0.05），导致生长速度减慢，导致该菌株的竞争力较野生菌株差。与大肠杆菌共培养时的典型菌株。 PFK1Δ菌株较低的种间竞争力进一步促进了共培养大肠杆菌的生长，进而激活了PFK1Δ菌株的乙醇生产效率，以对抗处于稳定期的大肠杆菌。研究结果阐明了PFK1基因对酿酒酵母生长和种间微生物竞争行为的调控，有助于我们了解酿酒酵母与其他微生物之间的微生物相互作用。