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    STUDIA CHEMIA - Issue no. 1 / 2018  
         
  Article:   HETEROLOGOUS EXPRESSION AND PURIFICATION OF RECOMBINANT CROTOXIN B, THE PHOSPHOLIPASE A2 SUBUNIT OF CROTOXIN.

Authors:  FRANCISC-ANDREI BODA, PÁL SALAMON, CSONGOR ORBÁN, LAVINIA BERTA, AUGUSTIN CURTICĂPEAN, ȘERBAN-ANDREI GÂZ, MARIA DOGARU.
 
       
         
  Abstract:  
DOI: 10.24193/subbchem.2018.1.01

Published Online: 2018-03-30
Published Print: 2018-03-30
pp. 7-20

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ABSTRACT: Crotoxin is a heterodimeric β-neurotoxin isolated from the venom of Crotalus durissus terrificus, consisting of two, non-covalently bound subunits, crotapotin (CA) and crotoxin B (CB). Both subunits present four different isoforms, consequently there are up to 16 different crotoxin complexes, each with different activity levels. The biological activities usually associated with crotoxin include neurotoxicity, myotoxicity and cardiotoxicity, however several other important biochemical and pharmacological effects of crotoxin have been observed, such as the antibacterial, antiviral, anti-inflammatory, antitumoral and analgesic activity. In this study we present the production of recombinant crotoxin B (isoform C). Expression of the protein was carried out using E. coli RosettaTM (DE3)pLysS strain, and the obtained protein was separated and purified using Ni2+-affinity chromatography. To the best of our knowledge the developed method represents the first reported method for obtaining the crotoxin B (isoform C) through heterologous expression using an efficient and cost-effective procedure.

Keywords: Crotoxin, neurotoxin, phospholipase A2, heterologous expression
 
         
     
         
         
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