Optimization of hyaluronic acid production through bacterial fermentation for industrial application

Abstract

This study aims to evaluate the potential for hyaluronic acid (HA) production and improve detection methods, using bacteria discovered by Dr. Bouzid Djihane, which have not yet been classified or identified. We performed several tests on these bacteria, including Gram staining, catalase testing, and India ink staining, to determine their characteristics and classify them. The bacteria were Gram-positive, appeared in chains, and possessed a capsule. To detect and purify hyaluronic acid, we used the Cetyltrimethylammonium bromide (CTAB) method and hyaluronidase enzyme assays. Purity tests using a NanoDrop spectrophotometer showed a 260/280 ratio of 2.2, indicating no protein contamination. The 260/230 ratio was approximately 0.9 for both commercial and bacterially produced HA, consistent across three replicates. Enzymatic degradation tests showed that the bacterial HA degraded slightly slower than commercial HA, possibly indicating a higher molecular weight. The specific activity of hyaluronidase on HA was confirmed, demonstrating its ability to cleave β-1,4-glycosidic bonds in the HA polymer. Importantly, this study confirmed that the newly discovered bacterial strain is capable of producing hyaluronic acid, and it provides a detailed description of improved HA production, detection, and purification methods, a confirmation that opens promising perspectives for biotechnological and medical applications.