overexpression and enzymatic assessment of antigenic fragments of hyaluronidase recombinant protein from streptococcus pyogenes

Background: hyaluronidase catalyzes the hydrolysis of hyaluronan polymers to n-acetyl-d-glucosamine and d-glucuronic acid. this enzyme is a dimer of identical subunits. hyaluronidase has different pharmaceutical and medical applications. previously, we produced a recombinant hyaluronidase antigenic fragment of streptococcus pyogenes. objectives: this study aimed to improve the protein production and purity of hyaluronidase recombinant protein from s. pyogenes. in addition, the enzymatic activity of this protein was investigated. materials and methods: the expression of hyaluronidase antigenic fragments was optimized using iptg concentration, time of induction, temperature, culture, and absorbance of 0.6-0.8-1 at 600 nm. afterwards, the expressed proteins were purified and the enzymatic activity was assessed by turbid metric method. results: data indicated that maximum protein is produced in od = 0.8, 0.5 mm isopropyl β-d-1-thiogalactopyranoside (iptg), 37ºc, nb 1.5x, without glucose, incubated for overnight. the enzymatic activity of the recombinant protein was similar to the commercial form of hyaluronidase. conclusions: the results showed that an antigenic fragment of the recombinant hyaluronidase protein from s. pyogenes has a considerable enzymatic activity. it can be suggested to use it for medical purposes. in addition, applications of bioinformatics software would facilitate the production of a smaller protein with same antigenic properties and enzymatic activity.