|چکیده انگلیسی||Overexpression and Enzymatic Assessment of Antigenic Fragments of Hyaluronidase Recombinant ProteinFrom Streptococcus pyogenes
Sadoogh Abbasian S1, Ghaznavi Rad E2, Akbari N3, Zolfaghari MR4, Pakzad I5, Abtahi H1.
• 1Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, IR Iran.
• 2Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
• 3Department of Microbiology, Faculty of Science, Arak branch, Islamic Azad University, Arak, IR Iran.
• 4Department of Microbiology, Qom Branch, Islamic Azad University, Qom, IR Iran.
• 5Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, IR Iran.
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 recombinanthyaluronidase antigenic fragment of Streptococcus pyogenes.
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.
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.
The results showed that an antigenic fragment of the recombinant hyaluronidase protein from S. pyogenes has a considerableenzymatic 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.
Enzymatic Activity; Hyaluronidase; Optimization of Expression|