INHIBITION OF E.CANCEROGENUS MAB-1 β -LACTAMASE BY SYNERGISTIC FORMULATION: AN IN VITRO HERBAL SHOTGUN STUDY

β-lactamases are increasing worldwide speedily by creating problem to currently available UTI therapies and hence there is an urgent need of research to find the effective therapy. In the present study we propose to develop drug formulation for the reversal of drug resistance in MDR E.cancerogenus MAB-1 by using bioenhancer property of isolated different phytochemicals (lead compounds). β-lactamase enzyme of E.cancerogenus MAB-1 was produced and purified up to 92.5 fold, by using Ion-Exchange chromatography(DEAE-Cellulose -52) techniques with 80.23 U/mg of specific activity. Iso-electric focusing and Sodium Dodecyl Sulfate ?Polyacrylamide gel electrophoresis (SDS-PAGE) study revealed enzyme with PI value 8.5 with Molecular weight ~6.5 KDa. Threonine (9.27 ppm) and Serine (3.94 ppm) were detected in highest quantity when analyzed by GC-MS technique. Organic and Water extracts of selected plants were investigated for GC-MS, IR, NMR analysis for the isolation of pure lead compounds i.e. Theobromine, Myricetin, Gallic acid, Ellagic acid, Kaempferol and Genistein. In vitro Enterobacter cancerogenus MAB-1 β-lactamase inhibition by Dug-phytochemical combinations was studied by Checker board and Time-Kill curve at their ? to 1/32 MIC values. The Theobromine+ Cefaclor synergy inhibited β-lactamases by 91%, Myricetin +Cefaclore (90%), Genistein+Cefaclore (92%), Genistein + Cefotaxime(90%), Genistein+Penicillin (91%) , Ellagic acid+Cefaclor (87%), Kaempferol+Ceftriaxone(87%), Kaempferol+Cefaclor (90%) and Clavulanic Acid (96%) Competitively, Gallic acid+Cefotaxime (92%) Non-Competitively. Thus the resulting data would be helpful to pick up the most effective and suitable combination therapy for further research in the area of drug development against UTI caused by E.cancerogenus MAB-1 or any member of Enterobacteriaceae family.