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Relative gene expression of bile salt hydrolase and surface proteins in two putative indigenous Lactobacillus plantarum strains under in vitro gut conditions

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Abstract

Probiotic bacteria must overcome the toxicity of bile salts secreted in the gut and adhere to the epithelial cells to enable their better colonization with extended transit time. Expression of bile salt hydrolase and other proteins on the surface of probiotic bacteria can help in better survivability and optimal functionality in the gut. Two putative Lactobacillus plantarum isolates i.e., Lp9 and Lp91 along with standard strain CSCC5276 were used. A battery of six housekeeping genes viz. gapB, dnaG, gyrA, ldhD, rpoD and 16S rRNA were evaluated by using geNorm 3.4 excel based application for normalizing the expression of bile salt hydrolase (bsh), mucus-binding protein (mub), mucus adhesion promoting protein (mapA), and elongation factor thermo unstable (EF-Tu) in Lp9 and Lp91. The maximal level of relative bsh gene expression was recorded in Lp91 with 2.89 ± 0.14, 4.57 ± 0.37 and 6.38 ± 0.19 fold increase at 2% bile salt concentration after 1, 2 and 3 h, respectively. Similarly, mub and mapA genes were maximally expressed in Lp9 at the level of 20.07 ± 1.28 and 30.92 ± 1.51 fold, when MRS was supplemented with 0.05% mucin and 1% each of bile and pancreatin (pH 6.5). However, in case of EF-Tu, the maximal expression of 42.84 ± 5.64 fold was recorded in Lp91 in the presence of mucin alone (0.05%). Hence, the expression of bsh, mub, mapA and EF-Tu could be considered as prospective biomarkers for screening of novel probiotic lactobacillus strains for optimal functionality in the gut.

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Acknowledgements

We gratefully acknowledge the Director, National Dairy Research Institute (NDRI, Karnal, India) for providing facilities to carry out the study. We thank Dr. N.·P. Shah (Australia) for kindly providing the standard Lactobacillus strain. The financial support received from Indian Council of Agricultural Research (ICAR, India) in terms of providing fellowship to the first author of the paper to carry out his doctoral program is greatly appreciated.

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Authors and Affiliations

  1. Molecular Biology Unit, Department of Dairy Microbiology, National Dairy Research Institute, Karnal, Haryana, 132001, India

    Virender Kumar Batish & Sunita Grover

  2. Department of Food Processing and Technology, Tezpur University, Napaam, Assam, 784028, India

    Raj Kumar Duary

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  1. Raj Kumar Duary
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  2. Virender Kumar Batish
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  3. Sunita Grover
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Correspondence to Sunita Grover.

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Duary, R.K., Batish, V.K. & Grover, S. Relative gene expression of bile salt hydrolase and surface proteins in two putative indigenous Lactobacillus plantarum strains under in vitro gut conditions. Mol Biol Rep 39, 2541–2552 (2012). https://doi.org/10.1007/s11033-011-1006-9

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