Extraction and purification of L-Asparaginase, L-Glutaminase from two Capsicum annuum varieties and their molecular characterization

Capsicum annuum L. varieties (Green and Red Chilly) were screened for the production of two antitumor enzymes; L-Asparaginase and L-Glutaminase. The crude protein concentration and the crude enzyme activities in both the varieties were determined. The enzymes were subjected to partial purification by anion exchange chromatography and the concentration of protein and the enzyme activities in purified L-Asparaginase and L-Glutaminase fractions were again determined. Both the enzyme fractions showed more L-Glutaminase activity than L-Asparaginase activity. L-Glutaminase activity was between 3 to 4 U/g and L-Asparaginase activity was between 2 to 3 U/g. Enzyme characterization studies showed that optimum activity of all enzyme fractions was at 37^oC, pH 9 and substrate concentration of 0.0006 g/l. Effect of activator and inhibitor concentration on enzyme activity were also studied. The molecular characterization of Green and Red chilly done by RFLP and RAPD analyses revealed that there was not much genetic heterogeneity between both the varieties of chilly.