Purification and Characterization of Hyaluronic Acid Produced by Priestia flexa N7

Abstract

Background: Hyaluronic acid (HA), a disaccharide polysaccharide, is widely used in medicine and cosmetics, with its applications depending on molecular weight (MW). Traditionally sourced from animals such as umbilical cords and chicken combs, extraction yields are low and cost high. Microbial fermentation offers a better alternative. This study aims to optimize the purification and characterization of HA from Priestia flexa N7 strain fermentation broth. Methods: The biochemical methods used in this study included Superdex™ G100 columns chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis, protein and nucleic acid quantitation assays, and purification of HA. Results: In this study, a method for purifying HA from P. flexa N7 fermentation broth was presented. Before purification, the crude HA was purified of toxins and low MW proteins using a mixture of activated charcoal (3%) with gamma alumina (2%) at a temperature of 25°C. Under these conditions, the yield of HA was 1720 mg/mL. The purification steps of HA were performed using adsorb toxins, ultrafiltration 50 kDa, and Superdex™ G100 columns. Purification efficiency is 38.5% with a purity of up to 7.5-fold after passing through Superdex™ G100 columns. The MW of HA as determined by SDS-PAGE was 459.6 kDa. Finally, the yield of HA was 11.89 mg/mL when precipitated with ammonium sulfate (60%). Conclusion: The present study describes a simple, economical process with higher yields when compared to other methods for the separation and purification of HA from P. flexa N7. The HA purification method in this study provides an opportunity to produce HA at a low cost and high yield.