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嗜酸乳杆菌在牛奶中生长期间的时间基因表达及益生菌特性

Temporal gene expression and probiotic attributes of Lactobacillus acidophilus during growth in milk.

作者信息

Azcarate-Peril M A, Tallon R, Klaenhammer T R

机构信息

Department of Food, Bioprocessing, and Nutrition Sciences, and Southeast Dairy Foods Research Center, North Carolina State University, Raleigh, NC 27695, USA.

出版信息

J Dairy Sci. 2009 Mar;92(3):870-86. doi: 10.3168/jds.2008-1457.

Abstract

Lactic acid bacteria have been used as starter strains in the production of fermented dairy products for centuries. Lactobacillus acidophilus is a widely recognized probiotic bacteria commonly added to yogurt and used in dietary supplements. In this study, a whole genome microarray was employed to monitor gene expression of L. acidophilus NCFM cells propagated in 11% skim milk during early, mid and late logarithmic phase, and stationary phase. Approximately 21% of 1,864 open reading frames were differentially expressed at least in one time point. Genes differentially expressed in skim milk included several members of the proteolytic enzyme system. Expression of prtP (proteinase precursor) and prtM (maturase) increased over time as well as several peptidases and transport systems. Expression of Opp1 (oligopeptide transport system 1) was highest at 4 h, whereas gene expression of Opp2 increased over time reaching its highest level at 12 h, suggesting that the 2 systems have different specificities. Expression of a 2-component regulatory system, previously shown to regulate acid tolerance and proteolytic activity, also increased during the early log and early stationary phases of growth. Expression of the genes involved in lactose utilization increased immediately (5 min) upon exposure to milk. The acidification activity, survival under storage conditions, and adhesion to mucin and Caco-2 tissue culture cells of selected mutants containing insertionally inactivated genes differentially expressed in the wild-type strain during growth in milk were examined for any potential links between probiotic properties and bacterial growth and survival in milk. Some of the most interesting genes found to be expressed in milk were correlated with signaling (autoinducer-2) and adherence to mucin and intestinal epithelial cells, in vitro.

摘要

几个世纪以来,乳酸菌一直被用作发酵乳制品生产的起始菌株。嗜酸乳杆菌是一种广为人知的益生菌,通常添加到酸奶中并用于膳食补充剂。在本研究中,采用全基因组微阵列来监测嗜酸乳杆菌NCFM细胞在11%脱脂乳中对数生长期早期、中期、晚期及稳定期的基因表达。在1864个开放阅读框中,约21%至少在一个时间点存在差异表达。在脱脂乳中差异表达的基因包括蛋白水解酶系统的几个成员。prtP(蛋白酶前体)和prtM(成熟酶)的表达随时间增加,一些肽酶和转运系统也是如此。Opp1(寡肽转运系统1)的表达在4小时时最高,而Opp2的基因表达随时间增加,在12小时时达到最高水平,这表明这两个系统具有不同的特异性。先前已证明可调节耐酸性和蛋白水解活性的双组分调节系统的表达,在生长的对数早期和稳定早期阶段也增加。参与乳糖利用的基因在接触牛奶后立即(5分钟)表达增加。对在牛奶中生长期间野生型菌株中差异表达的插入失活基因的选定突变体的酸化活性、储存条件下的存活率以及对粘蛋白和Caco-2组织培养细胞的粘附性进行了检测,以探究益生菌特性与细菌在牛奶中的生长和存活之间的潜在联系。一些在牛奶中表达的最有趣的基因与信号传导(自诱导物-2)以及体外对粘蛋白和肠上皮细胞的粘附相关。

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