Purification and Characterization of Hyaluronidase From <i>Streptomyces graminisoli</i>

Abstract

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.