Abstract

EXTRACTION OF POLYSACCHARIDE HYDROLYSATE FROM BACILLUS SUBTILIS IT'S ANTIOXIDANT ANTIBACTERIAL AND DNA DAMAGE STUDIES

S. Swetha, A. Dhinek, R. Ragunathan, Abirami V. and Jesteena Johney*

ABSTRACT

The present study investigates the production, characterization, and bioactivity of extracellular polyhydrolysate from Bacillus subtilis. Biomass estimation revealed a yield of 0.045 g, confirming effective bacterial growth and pellet extraction. Reducing sugar analysis using the DNS method demonstrated significant sugar content, with Ethanol treated sample exhibiting higher glucose concentration (81mg/ml) than the other, indicating enhanced polysaccharide hydrolysis. Thin Layer Chromatography (TLC) analysis showed distinct retention factor (Rf) values of 0.28 and 0.406 for Ethyl acetate and ethanol-treated samples, respectively, signifying variations in polyhydrolysate composition based on extraction methods. The polyhydrolysate exhibited notable antioxidant potential, with an activity of 187mg/ml, reinforcing its capacity to combat oxidative stress. Additionally, H2O2 scavenging assay results demonstrated a significant activities, confirming strongantioxidant properties. Antimicrobial activity analysis further highlighted inhibitory effects against test pathogens. Bacterial DNA isolation, involving centrifugation, enzymatic digestion, and organic solvent extraction, successfully yielded purified genomic DNA. Subsequent gel electrophoresis analysis revealed distinct banding patterns. Control DNA exhibited a sharp, intact band indicating genomic stability, whereas ethanol-treated DNA displayed smeared patterns, indicating fragmentation due to oxidative stress or direct ethanol-induced nucleic acid damage. Overall, the study confirms the extracellular polyhydrolysate's significant antioxidant and antimicrobial activities, emphasizing its potential as a promising bioactive compound for nutraceutical and therapeutic applications.

Keywords: Thin Layer Chromatography, Antioxidant Activity, H2O2 Scavenging Assay, DNA Damage Analysis, Nutraceutical Applications.