title: Overexpression and Enzymatic Assessment of Antigenic Fragments of Hyaluronidase Recombinant Protein From Streptococcus pyogenes.
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.
  
  
  OBJECTIVE
  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.
  
  
  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.
authors: Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, IR Iran.
authors: Shabnam
authors: S
authors: Sadoogh Abbasian
authors: Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
authors: Ehsanollah
authors: E
authors: Ghaznavi Rad
authors: Department of Microbiology, Faculty of Science, Arak branch, Islamic Azad University, Arak, IR Iran.
authors: Neda
authors: N
authors: Akbari
authors: Department of Microbiology, Qom Branch, Islamic Azad University, Qom, IR Iran.
authors: Mohammad Reza
authors: MR
authors: Zolfaghari
authors: Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, IR Iran.
authors: Iraj
authors: I
authors: Pakzad
authors: Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, IR Iran.
authors: Hamid
authors: H
authors: Abtahi
keywords: Enzymatic Activity; Hyaluronidase; Optimization of Expression