An alternative method to isolate protease and phospholipase A2 toxins from snake venoms based on partitioning of aqueous two-phase systems

Gómez GN (1), Nerli BB (2), Acosta OC (3), Picó GA (2), Leiva LCA (1)

(1) Protein Research Laboratory (LabInPro), Biochemistry Department, School of Natural and Exact Sciences, Northeast National University (UNNE), Corrientes, Argentina; (2) Laboratory of Physical Chemistry Applied to Bioseparation Processes, Department of Physical Chemistry, School of Biochemical and Pharmaceutical Sciences, National University of Rosario, Rosario, Argentina; (3) School of Veterinary Science, Northeast National University (UNNE), Corrientes, Argentina.

Abstract: Snake venoms are rich sources of active proteins that have been employed in the diagnosis and treatment of health disorders and antivenom therapy. Developing countries demand fast economical downstream processes for the purification of this biomolecule type without requiring sophisticated equipment. We developed an alternative, simple and easy to scale-up method, able to purify simultaneously protease and phospholipase A2 toxins from Bothrops alternatus venom. It comprises a multiple-step partition procedure with polyethylene-glycol/phosphate aqueous two-phase systems followed by a gel filtration chromatographic step. Two single bands in SDS-polyacrylamide gel electrophoresis and increased proteolytic and phospholipase A2 specific activities evidence the homogeneity of the isolated proteins.

Key words: partition, snake toxins, isolation, phospholipase A2, proteases.

ACKNOWLEDGMENTS

The authors would like to thank Laura Rey for supplying Bothrops alternatus venom (Serpentarium of the local zoo, Corrientes, Argentina). This work was financially supported by General Secretariat of Sciences and Technology, Northeast National University, Argentina (Projects n. PI F009/08 and PI F 018/10). G. N. Gómez is the recipient of a fellowship from General Secretariat of Sciences and Technology, Northeast National University, Argentina.

COPYRIGHT

© CEVAP 2012

SUBMISSION STATUS

Received: April 4, 2012.

Accepted: June 28, 2012.

Abstract published online: June 29, 2012.

Full paper published online: August 31, 2012.

CONFLICTS OF INTEREST

The authors declare no conflicts of interest.

FINANCIAL SOURCE

General Secretariat of Sciences and Technology - UNNE (PI F009/08 and PI F 018/10) provided the financial grants.

CORRESPONDENCE TO

Laura C. A. Leiva, Departamento de Bioquímica, Facultad de Ciencias Exactas y Naturales y Agrimensura (FaCENA), Universidad Nacional del Nordeste (UNNE), Av. Libertad 5470, (3400) Corrientes, Argentina. Phone: +54 0379 4457996 ext. 112. Email: lcleiva@exa.unne.edu.ar.