Dataset Information

Global Transcriptomic Analysis and Function Identification of Malolactic Enzyme Pathway of Lactobacillus paracasei L9 in Response to Bile Stress.

ABSTRACT: Tolerance to bile stress is crucial for Lactobacillus paracasei to survive in the intestinal tract and exert beneficial actions. In this work, global transcriptomic analysis revealed that 104 genes were significantly changed (log2FoldChange > 1.5, P < 0.05) in detected transcripts of L. paracasei L9 when exposed to 0.13% Ox-bile. The different expressed genes involved in various biological processes, including carbon source utilization, amino acids and peptide metabolism processes, transmembrane transport, transcription factors, and membrane proteins. It is noteworthy that gene mleS encoding malolactic enzyme (MLE) was 2.60-fold up-regulated. Meanwhile, L-malic acid was proved to enhance bile tolerance, which could be attributed to the intracellular alkalinization caused by MLE pathway. In addition, membrane vesicles were observed under bile stress, suggesting a disturbance in membrane charge without L-malic acid. Then, genetic and physiological experiments revealed that MLE pathway enhanced the bile tolerance by maintaining a membrane balance in L. paracasei L9, which will provide new insight into the molecular basis of MLE pathway involved in bile stress response in Lactic acid bacteria.

SUBMITTER: Ma X

PROVIDER: S-EPMC6119781 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Global Transcriptomic Analysis and Function Identification of Malolactic Enzyme Pathway of Lactobacillus paracasei L9 in Response to Bile Stress.

Ma Xiayin X Wang Guohong G Zhai Zhengyuan Z Zhou Pengyu P Hao Yanling Y

Frontiers in microbiology 20180823

Tolerance to bile stress is crucial for Lactobacillus paracasei to survive in the intestinal tract and exert beneficial actions. In this work, global transcriptomic analysis revealed that 104 genes were significantly changed (log2FoldChange > 1.5, P < 0.05) in detected transcripts of L. paracasei L9 when exposed to 0.13% Ox-bile. The different expressed genes involved in various biological processes, including carbon source utilization, amino acids and peptide met ...[more]

PMID: 30210466

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Project description:Lactic acid bacteria (LAB) are associated with various plant, animal, and human niches and are also present in many fermented foods and beverages. Thus, they are subjected to several stress conditions and have developed advanced response mechanisms to resist, adapt, and grow. This work aimed to identify the genes involved in some stress adaptation mechanisms in LAB. For this purpose, global reverse genetics was applied by screening a library of 1287 Lactobacillus paracasei transposon mutants for mild monofactorial stresses. This library was submitted independently to heat (52°C, 30 min), ethanol (170 g.L-1, 30 min), salt (NaCl 0.8 M, 24 h), acid (pH 4.5, 24 h), and oxidative (2 mM H2O2, 24 h) perturbations which trigger mild monofactorial stresses compatible with bacterial adaptation. Stress sensitivity of mutants was determined either by evaluating viability using propidium iodide (PI) staining, or by following growth inhibition through turbidity measurement. The screening for heat and ethanol stresses lead respectively to the identification of 63 and 27 genes/putative promoters whose disruption lead to an increased sensitivity. Among them, 14 genes or putative promoters were common for both stresses. For salt, acid and oxidative stresses, respectively 8, 6, and 9 genes or putative promoters were identified as essential for adaptation to these unfavorable conditions, with only three genes common to at least two stresses. Then, RT-qPCR was performed on selected stress response genes identified by mutant screenings in order to evaluate if their expression was modified in response to stresses in the parental strain. Eleven genes (membrane, transposase, chaperone, nucleotide and carbohydrate metabolism, and hypothetical protein genes) were upregulated during stress adaptation for at least two stresses. Seven genes, encoding membrane functions, were upregulated in response to a specific stress and thus could represent potential transcriptomic biomarkers. The results highlights that most of the genes identified by global reverse genetics are specifically required in response to one stress and that they are not differentially transcribed during stress in the parental strain. Most of these genes have not been characterized as stress response genes and provide new insights into the adaptation of lactic acid bacteria to their environment.

Dataset Information

Global Transcriptomic Analysis and Function Identification of Malolactic Enzyme Pathway of Lactobacillus paracasei L9 in Response to Bile Stress.

Publications

Global Transcriptomic Analysis and Function Identification of Malolactic Enzyme Pathway of <i>Lactobacillus paracasei</i> L9 in Response to Bile Stress.

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