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OJVRTM

Online Journal of Veterinary Research©

Volume 21(7):410-427, 2017.


Method to conjugate L-Aspariginase with Carboxymethyl Dextran

Marjan Chahardahcherik, Mahboobeh Ashrafi, Mahmoud Aminlari*

aDepartment of Biochemistry, School of Veterinary Medicine, Shiraz University, Shiraz,  71345 Iran *Mahmoud Aminlari, E. mail: aminlari@shirazu.ac.ir.

 

ABSTRACT

 

Chahardahcherika M, Ashrafia M, Aminlaria M., Method to conjugate L-Aspariginase with Carboxymethyl Dextran, Onl J Vet Res., 21(7):410-427, 2017. L-asparaginase aminohydrolase is used to manage childhood acute leukemia and non-Hodgkin's lymphoma by catalyzing hydrolysis of asparagine to aspartic acid and ammonia. The enzyme also prevents formation of acrylamide in foods processed at high temperatures. A method to conjugate L-asparaginase with carboxymethyl dextran (CMD) to increase specific activity is described. Conjugation was performed at pH 7.2 or 8.5 at a CMD:asparaginase molar ratio of 85:1 for 2h at room temperature. Conjugation was determined by presence of free amino groups and SDS-PAGE. Findings suggested that conjugation with CMD increased specific activity of L-asparaginase 2-3 fold, and its Michaelis constant (Km) 2-7 fold.

 Key words: L-asparaginase, conjugation, carboxymethyl dextran, enzyme activity, kinetic properties.


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