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不同来源β-葡聚糖对婴儿双歧杆菌亚种生长的比较蛋白质组分析及其利用模型。

Comparative proteome analysis of Bifidobacterium longum subsp. infantis grown on β-glucans from different sources and a model for their utilization.

机构信息

Food and Nutritional Sciences Program, School of Life Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.

出版信息

J Agric Food Chem. 2013 May 8;61(18):4360-70. doi: 10.1021/jf400792j. Epub 2013 Apr 29.

DOI:10.1021/jf400792j
PMID:23577653
Abstract

Recent studies have demonstrated that β-glucans from different sources, which are considered as potential prebiotics, could enhance growth of bifidobacteria. To elucidate the metabolic pathway of β-glucans in the widely used probiotic B. longum subsp. infantis, a comparative proteomic analysis was carried out along with two-dimensional difference gel electrophoresis (2D-DIGE), real-time RT-PCR, and enzyme activity assay on samples obtained from cultures grown on β-glucans derived from barley, seaweed, and mushroom. Results showed that 77 spots were found to be differentially expressed among different cultures, and 17 of them were predicted to play a role in β-glucan catabolism, including ABC transporter for sugars, enolase, and phosphotransferase system protein. Among them, 6 genes encoding for 6 proteins were shown to be induced by β-glucans at the transcriptional level and had higher abundance. The enzyme activity assay detected intracellular glucanase activity present in the cultures grown on the β-glucans from seaweed and mushroom. On the basis of the above results, a model for catabolism of β-glucans in B. infantis is proposed as follows: β-glucan molecules in the medium are transported into the cell through the ABC (ATP-binding cassette) transport system and PTS (phosphotransferase system) proteins followed by hydrolysis through action of intracellular glucanase to glucose, which is subsequently incorporated into the central fermentative pathway 'bifid shunt'. This study for the first time reveals the possible degradation pathway of β-glucans by B. infantis, which has implications for potential use of these β-glucans as novel prebiotics in development of synbiotic application.

摘要

最近的研究表明,来自不同来源的β-葡聚糖被认为是潜在的益生元,可以促进双歧杆菌的生长。为了阐明广泛使用的益生菌长双歧杆菌亚种婴儿双歧杆菌中β-葡聚糖的代谢途径,对从以大麦、海藻和蘑菇来源的β-葡聚糖培养的样品进行了比较蛋白质组学分析,包括二维差异凝胶电泳(2D-DIGE)、实时 RT-PCR 和酶活性测定。结果表明,在不同培养物中发现了 77 个差异表达的斑点,其中 17 个被预测在β-葡聚糖代谢中发挥作用,包括糖的 ABC 转运体、烯醇酶和磷酸转移酶系统蛋白。其中,有 6 个基因编码的 6 种蛋白质在转录水平上被β-葡聚糖诱导,且丰度较高。酶活性测定检测到在以海藻和蘑菇来源的β-葡聚糖培养的培养物中存在的细胞内葡聚糖酶活性。基于上述结果,提出了婴儿双歧杆菌β-葡聚糖代谢的模型如下:培养基中的β-葡聚糖分子通过 ABC(ATP 结合盒)转运系统和 PTS(磷酸转移酶系统)蛋白进入细胞,然后通过细胞内葡聚糖酶水解成葡萄糖,随后葡萄糖被纳入中央发酵途径“双歧支路”。本研究首次揭示了婴儿双歧杆菌降解β-葡聚糖的可能途径,这对于将这些β-葡聚糖作为新型益生元应用于共生体的开发具有重要意义。

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