Equipe de Chimie Analytique des Molécules Bio-Actives, IPHC-DSA, Université de Strasbourg, CNRS, 67400, Illkirch-Graffenstaden, France.
J Proteome Res. 2012 Jan 1;11(1):109-18. doi: 10.1021/pr200828t. Epub 2011 Nov 17.
The identification of cell determinants involved in probiotic features is a challenge in current probiotic research. In this work, markers of bile tolerance in Lactobacillus casei were investigated using comparative proteomics. Six L. casei strains were classified on the basis of their ability to grow in the presence of bile salts in vitro. Constitutive differences between whole cell proteomes of the most tolerant strain (L. casei Rosell-215), the most sensitive one (L. casei ATCC 334), and a moderately tolerant strain (L. casei DN-114 001) were investigated. The ascertained subproteome was further studied for the six strains in both standard and bile stressing conditions. Focus was on proteins whose expression levels were correlated with observed levels of bile tolerance in vitro, particularly those previously reported to be involved in the bile tolerance process of lactobacilli. Analysis revealed that 12 proteins involved in membrane modification (NagA, NagB, and RmlC), cell protection and detoxification (ClpL and OpuA), as well as central metabolism (Eno, GndA, Pgm, Pta, Pyk, Rp1l, and ThRS) were likely to be key determinants of bile tolerance in L. casei and may serve as potential biomarkers for phenotyping or screening purposes. The approach used enabled the correlation of expression levels of particular proteins with a specific probiotic trait.
确定参与益生菌特性的细胞决定因素是当前益生菌研究中的一个挑战。在这项工作中,使用比较蛋白质组学研究了干酪乳杆菌耐胆盐的标记物。根据体外胆汁盐存在下生长的能力,将 6 株干酪乳杆菌菌株进行分类。研究了最耐受菌株(干酪乳杆菌 Rosell-215)、最敏感菌株(干酪乳杆菌 ATCC 334)和中度耐受菌株(干酪乳杆菌 DN-114 001)的全细胞蛋白质组之间的组成差异。在标准和胆汁应激条件下,进一步研究了这 6 株菌株的确定亚蛋白质组。重点是那些表达水平与体外观察到的胆汁耐受性相关的蛋白质,特别是那些先前报道与乳杆菌胆汁耐受性过程有关的蛋白质。分析表明,12 种涉及膜修饰(NagA、NagB 和 RmlC)、细胞保护和解毒(ClpL 和 OpuA)以及中心代谢(Eno、GndA、Pgm、Pta、Pyk、Rp1l 和 ThRS)的蛋白质可能是干酪乳杆菌耐胆盐的关键决定因素,可作为表型或筛选目的的潜在生物标志物。所使用的方法能够将特定蛋白质的表达水平与特定的益生菌特性相关联。