Abstract
An Unique extracelluar glutaminase free L-asparaginase from novel marine Actinomycetes was isolated to perceptible homogeneity in agro industrial wastes. Quantitative Preparative Continuous-Elution SDS PAGE Electrophoresis is a high-resolution method for the preparative isolation of L-Asparaginase in biological samples. The enzyme was purified 248.68-fold and showed a final specific activity of 5035.28 IU/mg with an 80.71% yield. The homotetramer enzyme has a molecular mass of 133.25 kDa and an isoelectric point of approximately 5.4.Kinetic parameters, Km and Vmax of purified L-asparaginase from Streptomyces radiopugnans MS1 were found to be 0.0598, 3.5478 IU μg - 1 respectively. The de novo sequencing strategy presented here provides a rapid and reliable means to identify proteins in Streptomyces radiopugnans MS1. The purified L-asparaginase has no glutaminase activity, which can diminish the leeway of side effects during the itinerary of anti-malignancy therapy.
Highlights
L-asparaginase is used in the treatment of malignancies of the multiorgans [1]
Proportional examinations of preparations of E. coli, L-asparaginase produced in the USSR, Germany (Crasntin) and Japan (Leumase) were made [4] and it revealed that clinically USSR preparation was superior to that made in Japan
We report here a High yield method which can be used for both preparative and analytical scale purification of L-Asparaginase with sodium dodecyl sulfate (SDS) PAGE by using continuous elution electrophoresis
Summary
L-asparaginase is used in the treatment of malignancies of the multiorgans [1]. The use of L-asparaginase in anti-cancer therapy is based on its ability to catalyze the conversion of L-Asparagine to L-Aspartate and ammonia in human body fluid. Enormous numbers of researches have been accomplished on the biosynthesis of this therapeutically important L-Asparaginase, since Mashburn and Wriston demonstrated anti tumor activity [3]. The FDA and WHO organizations have approved L-asparaginase for the successful treatment of Acute Lymphoblastic Leukemia (ALL) and Lymphsarcoma. Proportional examinations of preparations of E. coli, L-asparaginase produced in the USSR, Germany (Crasntin) and Japan (Leumase) were made [4] and it revealed that clinically USSR preparation was superior to that made in Japan
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