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比较生理学和转录组学分析超声生化控制对德氏乳杆菌保加利亚亚种后酸化的影响。

Comparative physiological and transcriptomic analysis of sono-biochemical control over post-acidification of Lactobacillus delbrueckii subsp. bulgaricus.

机构信息

College of Food Science and Engineering, Zhejiang Key Laboratory of Intelligent Food Logistic and Processing, Zhejiang-Malaysia Joint Research Laboratory for Agricultural Product Processing and Nutrition, Ningbo University, Ningbo, 315211, China.

State Key Laboratory of Dairy Biotechnology, Shanghai Engineering Research Center of Dairy Biotechnology, Dairy Research Institute, Bright Dairy & Food Co., Ltd., Shanghai, 200436, China.

出版信息

Food Microbiol. 2024 Sep;122:104563. doi: 10.1016/j.fm.2024.104563. Epub 2024 May 18.

Abstract

Thermosonication (UT) prestress treatments combining with varied fermentation patterns has been revealed as an effective method to regulate post-acidification as exerted by Lactobacillus delbrueckii subsp. bulgaricus (L. delbrueckii), but sono-biochemical controlling mechanisms remain elusive. This study employed physiological and transcriptomic analysis to explore the response mechanism of L. delbrueckii to UT-induced microstress (600 W, 33 kHz, 10 min). UT stress-induced inhibition of acidification of L. delbrueckii during (post)-fermentation was first confirmed, relying on the UT process parameters such as stress exposure duration and UT power. The significantly enhanced membrane permeability in cells treated by 600 W for 10 min than the microbes stressed by 420 W for 20 min suggested the higher dependence of UT-derived stresses on the treatment durations, relative to the ultrasonic powers. In addition, ultrasonication treatment-induced changes in cell membrane integrity enhanced and/or disrupted permeability of L. delbrueckii, resulting in an imbalance in intracellular conditions associated with corresponding alterations in metabolic behaviors and fermentation efficiencies. UT-prestressed inoculum exhibited a 21.46% decrease in the membrane potential during the lag phase compared to untreated samples, with an intracellular pH of 5.68 ± 0.12, attributed to the lower activities of H-ATPase and lactate dehydrogenase due to UT stress pretreatments. Comparative transcriptomic analysis revealed that UT prestress influenced the genes related to glycolysis, pyruvate metabolism, fatty acid synthesis, and ABC transport. The genes encoding 3-oxoacyl-[acyl-carrier-protein] reductases I, II, and III, CoA carboxylase, lactate dehydrogenase, pyruvate oxidase, glucose-6-phosphate isomerase, and glycerol-3-phosphate dehydrogenase were downregulated, thus identifying the relevance of the UT microstresses-downregulated absorption and utilization of carbohydrates with the attenuated fatty acid production and energy metabolisms. These findings could contribute to provide a better understanding of the inactivated effects on the post-acidification of L. delbrueckii by ultrasonic pretreatments, thus providing theoretical basis for the targeted optimization of acidification inhibition efficiencies for yogurt products during chilled preservation processes.

摘要

声处理(UT)预应力处理与不同的发酵模式相结合,已被证明是一种有效调节保加利亚乳杆菌(Lactobacillus delbrueckii subsp. bulgaricus,L. delbrueckii)后酸化的方法,但声生化控制机制仍不清楚。本研究采用生理和转录组分析方法,探讨了 L. delbrueckii 对 UT 诱导微应激(600 W、33 kHz、10 min)的响应机制。首先证实了 UT 应激抑制了发酵后(后)L. delbrueckii 的酸化,这依赖于 UT 过程参数,如应激暴露持续时间和 UT 功率。与用 420 W 处理 20 min 的微生物相比,用 600 W 处理 10 min 的细胞的膜通透性显著增强,这表明 UT 衍生的应激对处理时间的依赖性高于超声功率。此外,超声处理诱导的细胞膜完整性变化增强和/或破坏了 L. delbrueckii 的通透性,导致与代谢行为和发酵效率变化相关的细胞内条件失衡。与未处理的样品相比,UT 预应激接种物在迟滞期的膜电位下降了 21.46%,细胞内 pH 值为 5.68±0.12,这归因于 UT 应激预处理导致 H-ATPase 和乳酸脱氢酶活性降低。比较转录组分析显示,UT 预处理影响与糖酵解、丙酮酸代谢、脂肪酸合成和 ABC 转运相关的基因。编码 3-氧酰基-[酰基载体蛋白]还原酶 I、II 和 III、辅酶 A 羧化酶、乳酸脱氢酶、丙酮酸氧化酶、葡萄糖-6-磷酸异构酶和甘油-3-磷酸脱氢酶的基因下调,因此确定了 UT 微应激下调碳水化合物的吸收和利用与减弱的脂肪酸生成和能量代谢之间的相关性。这些发现有助于更好地理解超声预处理对 L. delbrueckii 后酸化的灭活作用,从而为冷藏保存过程中酸奶产品酸化抑制效率的靶向优化提供理论依据。

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