Isolation and characterization of peptides from Gymnema sylvestre and their antimicrobial assay against bacterial isolates of diabetic foot ulcer

Abstract

Diabetic foot ulcers (DFUs) are significant micro vascular diabetes-related lesions that result from a number of predisposing factors, including infections, bone abnormalities, diabetic neuropathy and peripheral arterial disease. If left untreated, these conditions may exacerbate and ultimately result in lower limb amputation. The development of foot ulcers is a significant consequence associated with diabetes. The current global issue is the emergence and spread of antibiotic-resistant microorganisms. Therefore, it is imperative that alternative chemical development proceed immediately. Antimicrobial peptides are referred to as host defense peptides area component of the innate immune system present in every living organism. These small molecules play significant roles in the battle against infection through their wide-spectrum antibacterial action, host immune-modulatory capabilities and other aspects related to wound healing. The present study was carried out to isolate and characterize peptides having antibacterial activity from leaves of Gymnema sylvestre (Gurmar). Crude extract of leaves was prepared in Phosphate Buffer Saline (PBS) and antibacterial activity was checked on Luria Bertani (LB) broth agar plates against several bacteria such as Escherichia coli, Staphylococcus aureus, Klebsiella pneumonia, Pseudomonas aeruginosa and Enterococcus faciealis. The extract was used for peptide(s) precipitation with different percentage of ammonium sulfate solution, homogeneity and molecular mass of protein fractions was determined by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS–PAGE). Purification of isolated protein/peptide was done by Gel Filtration Chromatography (GFC) using Sephadex G-50 and characterization was done by Reverse Phase High Performance Liquid Chromatography (RP-HPLC).