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Functional Genomic Analyses of Exopolysaccharide-Producing Streptococcus thermophilus ASCC 1275 in Response to Milk Fermentation Conditions

机译:产外多糖的嗜热链球菌ASCC 1275对牛奶发酵条件的功能基因组分析

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摘要

Exopolysaccharide (EPS) produced from dairy bacteria improves texture and functionalities of fermented dairy foods. Our previous study showed improved EPS production from Streptococcus thermophilus ASCC1275 (ST1275) by simple alteration of fermentation conditions such as pH decrease (pH 6.5 → pH 5.5), temperature increase (37°C → 40°C) and/or whey protein isolate (WPI) supplementation. The iTRAQ-based proteomics in combination with transcriptomics were applied to understand cellular protein expression in ST1275 in response to above shifts during milk fermentation. The pH decrease induced the most differentially expressed proteins (DEPs) that are involved in cellular metabolic responses including glutamate catabolism, arginine biosynthesis, cysteine catabolism, purine metabolism, lactose uptake, and fatty acid biosynthesis. Temperature increase and WPI supplementation did not induce much changes in global protein express profiles of ST1275 between comparisons of pH 5.5 conditions. Comparative proteomic analyses from pairwise comparisons demonstrated enhanced glutamate catabolism and purine metabolism under pH 5.5 conditions (Cd2, Cd3, and Cd4) compared to that of pH 6.5 condition (Cd1). Concordance analysis for differential expressed genes (DEGs) and DEPs highlighted down-regulated glutamate catabolism and up-regulated arginine biosynthesis in pH 5.5 conditions. Down regulation of glutamate catabolism was also confirmed by pathway enrichment analysis. Down-regulation of EpsB involved in EPS assembly was observed at both mRNA and protein level in pH 5.5 conditions compared to that in pH 6.5 condition. Medium pH decreased to mild acidic level induced cellular changes associated with glutamate catabolism, arginine biosynthesis and regulation of EPS assembly in ST1275.
机译:乳制品细菌产生的胞外多糖(EPS)改善了发酵乳制品的质地和功能。我们以前的研究表明,通过简单地改变发酵条件,例如pH降低(pH 6.5→pH 5.5),温度升高(37°C→40°C)和/或分离乳清蛋白,可以改善嗜热链球菌ASCC1275(ST1275)的EPS产量。 WPI)补充。基于iTRAQ的蛋白质组学与转录组学相结合,可用于了解牛奶发酵过程中上述变化对ST1275中细胞蛋白质表达的影响。 pH值降低会诱导参与细胞代谢反应的最差异表达蛋白质(DEP),这些代谢反应包括谷氨酸分解代谢,精氨酸的生物合成,半胱氨酸分解代谢,嘌呤代谢,乳糖摄取和脂肪酸生物合成。在pH 5.5条件的比较之间,温度升高和WPI补充并未在ST1275的整体蛋白表达谱中引起太多变化。通过成对比较进行的蛋白质组分析比较表明,与pH 6.5条件(Cd1)相比,在pH 5.5条件(Cd2,Cd3和Cd4)下谷氨酸分解代谢和嘌呤代谢增强。差异表达基因(DEGs)和DEPs的一致性分析突出显示了在pH 5.5条件下谷氨酸分解代谢的下调和精氨酸生物合成的上调。通过途径富集分析也证实了谷氨酸分解代谢的下调。与在pH 6.5条件下相比,在pH 5.5条件下在mRNA和蛋白质水平上均观察到参与EPS装配的EpsB的下调。在ST1275中,中等pH值降低至中等酸性水平会诱导与谷氨酸分解代谢,精氨酸生物合成和EPS装配调控相关的细胞变化。

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