Abstract

NEW RECOMBINATION OF LACTOBACILLUS ACIDOPHILUS BILE SALT HYDROLASE GENE A (BSHA) AND TRANSFORMATION IN ESCHERICHIA COLI

Atheer Ahmed*

Abstract

The bile salt hydrolase gene (bshA), encoding bile salt hydrolase enzyme (EC 3.5.1.24) from Lactobacillus acidophilus Ar probiotic isolate, which is responsible for assimilation cholesterol were studied in the present work. This gene designed contain two restriction sites (PstI/SacI) were added to each end of fragment and amplified by PCR techniques use specific primer to genetic analysis sequencing for gene nucleotides which were 801 bp. Gene manipulation during cloning inserted into pMGe36 vector respectively. pMGe36 vector is overexpression plasmid for E. coli. The resulted constructs were named as pMG36e / bshA. The recombinant was transferred to E. coli MC1022 by chemical transformation. Obtained recombinants analyzed for expression and sequences. The results were confirmed that production of bile salt hydrolase from recombinant E.coli MC1022 pMGe36/bshA found to be higher expression while compared with E.coli MC1022 wild type strain from 100 to 184.78 (U\mg). The recombinant plasmid also found to be stable in host organism after a few generations.

Keywords: Probiotic strain Lactobacillus acidophilus Ar, E. coli MC1022, Bile salt hydrolase, bshA genes.