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:: Volume 17, Number 4 (7-2014) ::
amuj 2014, 17(4): 67-75 Back to browse issues page
Production and Purification of Enzymatic Region of Streptococcal Hyaluronidase in E.coli
Nafise O Sadat Mirjamali Mehrabadi, Safieh Soufian, Hamid Abtahi
Associate Professor, Department of Microbiology, School of Medicine, Arak University of Medical Sciences, Arak, Iran , habtahi2001@yahoo.com
Abstract:   (4401 Views)

Background: Streptococcus pyogenes produce extracellular hyaluronidase enzyme which is directly associated with the spreading of the organism during infection. Hyaluronidase enzyme is able to break hyaluronic acid or interstitial cement. This enzyme might be used in cancer treatment.The objective of the present study was to clone and express the nucleotide sequence of this enzyme which is involved in hyaluronidase enzymatic activity.

Materials and Methods: The enzymatic region of hyaluronidase gene was detected by bioinformatics methods. The polymerase chain reaction method was used to amplify the region. The amplified product was cloned into the expression vector pET32a. E. coli BL21 (DE3) pLYsS was transformed with recombinant plasmids. Then gene expression was induced by IPTG. The expressed protein was purified successfully via affinity chromatography by NiNTA kit. The integrity of the product was confirmed by western-blot analysis.

Results: The nucleotide sequence of amplified gene was consistent with the streptocuccal hyaluronidase gene. The concentration of recombinant protein calculated to 500 mg purified protein per liter. The enzymatic region of recombinant protein from Streptococcus pyogenes was recognized by all five patient’s sera with Streptococcus infection.

Conclusion: In general, it is possible to produce the enzymatic regions of the Streptococcus pyogenes hyaluronidase in Escherichia coli. The antigenic property of the produced protein is well retained. Considering the product's domestic demand and also low efficiency of production and pathogenicity of Streptococcus species, it is possible to produce it as recombinant product.

Keywords: Cloning, Gene expression, Hyaluronidase, Streptococcus pyogenes
Full-Text [PDF 331 kb]   (1027 Downloads)    
Type of Study: Original Atricle | Subject: Basic Sciences
Received: 2014/03/11 | Accepted: 2014/05/7 | Published: 2014/06/21
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Mirjamali Mehrabadi N O S, Soufian S, Abtahi H. Production and Purification of Enzymatic Region of Streptococcal Hyaluronidase in E.coli. amuj. 2014; 17 (4) :67-75
URL: http://amuj.arakmu.ac.ir/article-1-2838-en.html
Volume 17, Number 4 (7-2014) Back to browse issues page
مجله دانشگاه علوم پزشکی اراک Arak Medical University Journal
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