title: Relative gene expression of bile salt hydrolase and surface proteins in two putative indigenous Lactobacillus plantarum strains under in vitro gut conditions.
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.
authors.affiliation: Department of Food Processing and Technology, Tezpur University, Napaam, Assam 784028, India.
authors.forename: Raj Kumar
authors.initials: RK
authors.lastname: Duary
authors.forename: Virender Kumar
authors.initials: VK
authors.lastname: Batish
authors.forename: Sunita
authors.initials: S
authors.lastname: Grover
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D001646:Bile; D018929:Cell Culture Techniques; D017931:DNA Primers / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D048191:Lactiplantibacillus plantarum / Q000201:enzymology*; D008565:Membrane Proteins / Q000378:metabolism*; D009077:Mucins; D010194:Pancreatin; D019936:Probiotics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D008565:Membrane Proteins; D009077:Mucins; D010194:Pancreatin; D000581:Amidohydrolases; C023154:choloylglycine hydrolase