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Development And Characterization Of Engineered Commensal Bacteria For The Delivery Of Enzymes

Author(s): Nitin K Jain, M.K Chourasia, S Jain, S.K Jain, N.K Jain

The present study was aimed at development and characterization of engineered commensal bacteria for site-specific targeting of enzymes. Separation of plasmid (pJR2 and GB4) and transformation in Lactobacillus acidophilus was carried out using a reported method. Azo polymers were synthesized using water-soluble and water- insoluble monomers belonging to acrylate series. The synthesized polymers were characterized for physical appearance, solubility and film forming properties. Effect of microbial flora of colon on polymers was seen by incubating the glass cover slips coated with azo polymers in sterilized nutrient broth containing freshly voided human fecal culture suspension. Colon-specific drug delivery system was developed by mixing the bacterial pellet with skimmed milk and then coating these granules with azo- and pH-sensitive polymer. Lowering of galactose concentration and a-oxoglutaric acid concentration were monitored to in vitro characterize E. coli (56) and E. coli(797), respectively. In vivo screening of developed oral, colon-specific formulation, azo polymer coated granules of engineered E. coli(56) producing galactokinase (GK) and engineered E. coli(797) producing glutamate dehydrogenase (GDH) was performed on rabbits. D-galactose, which is the substrate for enzyme galactokinase and ammonium chloride, was administered intraperitoneally to all rabbits. The lowering of blood galactoselevel and urea level was observed with respect to variable time interval.


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