Purification and Characterization of Hyaluronidase From Streptomyces graminisoli.

Abstract

Hyaluronidases (HAase) are a family of hydrolytic enzymes that degrade hyaluronic acid (HA) and have gained significant attention in medical and pharmaceutical applications. This study presents an optimized approach for isolating and characterizing a novel HAase-producing actinomycete strain. Using HA-supplemented minimal salt medium (MSM) as the sole carbon and nitrogen source, we isolated and identified Streptomyces graminisoli by 16S-rRNA sequencing. The enzyme was purified using an improved DEAE-Sephadex G50 column chromatography method, yielding a molecular weight of 35 kDa. Kinetic analysis results revealed a K_m value of 0.28 mg/mL and a V_max value of 24.9 U/mL, indicating high substrate affinity. The results of advanced bioinformatics analysis using both nucleotide (Basic Local Alignment Search Tool nucleotide [BLASTN]) and amino acid (Basic Local Alignment Search Tool protein [BLASTP]) sequences demonstrated significant conservation across actinomycetes, with sequence identity and query coverage exceeding 75% and 90%, respectively. This study provides new insights into microbial HAase production and characterization, with potential applications in biotechnology and medicine.