Molecular Cloning and Sequence Analysis of Bile Salt Hydrolase Gene (bsh) from Lactobacillus plantarum MBUL90 Strain of Human Origin

Abstract

This study provided genetic information on a bile salt hydrolase (bsh) of a Lactobacillus plantarum strain of Indian origin, MBUL90. L. plantarum strains were screened by PCR for the determination of the bsh locus in their genome using specific primers. None of the lactobacilli strains produced the expected size of amplicon (∼ 1.0 kb) except L. plantarum strains, which proved the specificity of the primers. The bsh amplicon of L. plantarum MBUL90 was cloned into pDrive vector, and nucleotide sequences were determined. Sequence analysis of bsh genes revealed a high level of similarity within the species of L. plantarum as well as with other species of Lactobacillus. The resulting nucleotide sequence of an ORF of 975 bp encoded a predicted protein of 324 amino acids representing a theoretical molecular mass of 37 kDa with a pI of 4.92. The protein deduced from the complete ORF had high similarity with other Bsh proteins, and four highly conserved amino acid motifs (YFGRNXD, NEXGLXXAGLNF, VXVLTNNPXF, and SXSRFVRXAF) were located around the active site. Genetic data presented in this paper provide a sound foundation for better understanding the genetic diversity of bsh in Lactobacillus genus and may provide a new genetic marker for phylogenetic study.

Keywords:

• bile salt hydrolase
• Lactobacillus plantarum
• molecular cloning
• PCR
• taurodeoxycholic acid

ACKNOWLEDGMENT

The authors gratefully acknowledge the financial support received from Department of Biotechnology, India. Mr. Rajesh Kumar is the recipient of scholarship from Indian Council of Agriculture Research and duly acknowledges the financial assistance received from the Council.