Genetic modification of Probiotic Lactobacillus by incorporation of Cathelicidin (LL-37), Colicin E2 and Inulosucrase (InuJ) gene

Abstract

Lactobacilli are widely used in probiotic preparations. They have not been associated with disease and for more than 100 years have been regarded as non-pathogenic members of the intestinal and urogenital floras. Lactobacilli are used in treatment of UTI but are inefficient in complete inhibition of the pathogen. The antimicrobial activity of these Lactobacilli can be increased by incorporating antimicrobial peptides such as Cathelicidin (LL-37) and Colicin E2. For this, cathelicidin (LL-37) and colicin E2 were amplified from pCDNA5/FRT and pCoLE2-P9 plasmid by PCR and ligated with pSLP111.3 plasmid (an E.coli-Lactobacillus shuttle vector). The increase consumption of refined carbohydrates and fat-enriched diet has led to an increase in the risk of insulin resistance. Inulosucrase enzyme produced by Lactobacillus present in the gastrointestinal tract synthesizes inulin which increases glucose uptake and decreases insulin resistance. The enzyme is produced in low amounts in the GI tract. By incorporating InuJ in probiotic Lactobacilli the production of inulosucrase enzyme in gastrointestinal tract can be increased. For this, InuJ was amplified from the plasmid pMAL-p2 by PCR and needs to be cloned in pSLP111.3 plasmid