Integrated Transcriptomic and Proteomic Analysis of the Bile Stress Response in a Centenarian-originated Probiotic Bifidobacterium longum BBMN68

被引:76
作者
An, Haoran [1 ,2 ]
Douillard, Francois P. [3 ]
Wang, Guohong [1 ,2 ]
Zhai, Zhengyuan [1 ,2 ]
Yang, Jin [4 ]
Song, Shuhui [4 ]
Cui, Jianyun [1 ,2 ]
Ren, Fazheng [1 ,2 ]
Luo, Yunbo [1 ,2 ]
Zhang, Bing [4 ]
Hao, Yanling [1 ,2 ]
机构
[1] China Agr Univ, Key Lab Funct Dairy, Minist Educ, Coll Food Sci & Nutr Engn, Beijing 100083, Peoples R China
[2] China Agr Univ, Key Lab Funct Dairy, Beijing Municipal, Coll Food Sci & Nutr Engn, Beijing 100083, Peoples R China
[3] Univ Helsinki, Dept Vet Biosci, Helsinki, Finland
[4] Chinese Acad Sci, Beijing Inst Genom, Core Genom Facil, Beijing 100101, Peoples R China
关键词
ENTEROCOCCUS-FAECALIS V583; ESCHERICHIA-COLI; HEAT-SHOCK; LACTOBACILLUS-REUTERI; LACTOCOCCUS-LACTIS; OXIDATIVE STRESS; SALT HYDROLASE; MOLECULAR CHARACTERIZATION; MECHANOSENSITIVE CHANNELS; FUNCTIONAL-ANALYSIS;
D O I
10.1074/mcp.M114.039156
中图分类号
Q5 [生物化学];
学科分类号
070307 [化学生物学];
摘要
Bifidobacteria are natural inhabitants of the human gastrointestinal tract and well known for their health-promoting effects. Tolerance to bile stress is crucial for bifidobacteria to survive in the colon and to exert their beneficial actions. In this work, RNA-Seq transcriptomic analysis complemented with proteomic analysis was used to investigate the cellular response to bile in Bifidobacterium longum BBMN68. The transcript levels of 236 genes were significantly changed ( threefold, p < 0.001) and 44 proteins were differentially abundant (1.6-fold, p < 0.01) in B. longum BBMN68 when exposed to 0.75 g l(-1) ox-bile. The hemolysin-like protein and bile efflux systems were significantly over produced, which might prevent bile adsorption and exclude bile, respectively. The cell membrane composition was modified probably by an increase of cyclopropane fatty acid and a decrease of transmembrane proteins, resulting in a cell membrane more impermeable to bile salts. Our hypothesis was later confirmed by surface hydrophobicity assay. The transcription of genes related to xylose utilization and bifid shunt were up-regulated, which increased the production of ATP and reducing equivalents to cope with bile-induced damages in a xylan-rich colon environment. Bile salts signal the B. longum BBMN68 to gut entrance and enhance the expression of esterase and sortase associated with adhesion and colonization in intestinal tract, which was supported by a fivefold increased adhesion ability to HT-29 cells by BBMN68 upon bile exposure. Notably, bacterial one-hybrid and EMSA assay revealed that the two-component system senX3-regX3 controlled the expression of pstS in bifidobacteria and the role of this target gene in bile resistance was further verified by heterologous expression in Lactococcus lactis. Taken altogether, this study established a model for global response mechanisms in B. longum to bile.
引用
收藏
页码:2558 / 2572
页数:15
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