Investigation of the in vitro effects of some antibiotics on the purified Beta-glucosidases from the rat liver

Abstract

Beta-glucosidases catalyzes the hydrolysis of the glycosidic bondsto terminal non-reducing residues inb-D-glucosides and oligosac-charides.b-glucosidases are widely distributed in the livingworld.b-glucosidases which in mammals, primarily found in theliver and kidneys;lysosomalb-glucosidase (GBA1),non-lysosomalb-glucosidase (GBA2), cytosolicb-glucosidase (GBA3),intestinallactase-phloriz the hydrolase (LPH).Liver tissues of Wistar-Albino rats were homogenized withhomogenizer in the extraction buffer and crude extract wasobtained after centrifugation.Ammoniumsulfate precipitationrange designated crude extract was purified by sepharose 4B-L-tyrosine-1-naphthylamine hydrophobic gel.Commercially avail-abled antibiotics were prepared with substrate buffer.It wasinvestigated inhbition effects of cefuroximesodium, ampicillin-sul-bactam, amoxicillin trihydrate/potassium clavulanate, cefazolinsodium, gentamicin sulfate and ceftriaxone disodium antibioticsonto GBA2.Inhibition types and Kivalues of related antibioticswere determined with p-NPG substrate.Lineweaver-Burk plotwas used for that purpose.Rat liver GBA2 was purified at 30.2-fold with 43.4%yield.GBA2 was illustrated 58 and 110 kDa at SDS-PAGE.IC50value of ampicillin/sulbactam antibiotic for GBA2 was found62.97 mg/ml with competitive type inhibition and other antibi-otics didn’t inhibit.Purfication methods are being used in the literature for thepurifiedb-glucosidase from different sources.Purified GBA2 wasillustrated 58 and 110 kDa at SDS-PAGE.About molecularweight ofb- glucosidases is presented different information in theliterat€ure. This has been reported because of acid beta glucosidases are abnormal migration at the acrylamide or agarosegels.It was investigated inhbition effects of various antibioticsonto purified GBA2.Ampicillin/sulbactam antibiotic inhibited topurfied GBA2 at the competitive type.Similiar antibiotics studieshave been made in the literature for different enzymes.