Author/Authors :
Sadoogh Abbasian، Shabnam نويسنده Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, IR Iran , , Ghaznavi-Rad، Ehsanollah نويسنده Department of microbiology and immunology, Arak University of Medical Sciences, Arak, Iran. Ghaznavi-Rad, Ehsanollah , Akbari، Neda نويسنده Biochemistry Department, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran , , Zolfaghari، Mohammad Reza نويسنده , , Pakzad، Iraj نويسنده Department of Microbiology and Clinical Microbiology Research Center, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, IR Iran , , Abtahi، Hamid نويسنده Department of Microbiology, Molecular and Medicine Research Center, Arak University of Medical Sciences Abtahi, Hamid
Abstract :
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 B-D-1-thiogalactopyranoside (IPTG), 37?C, 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.
