Abstract
To screen soil metagenomic libraries for novel enzymes with enhanced activities. To screen soil metagenomic libraries for novel enzymes with enhanced activities. A novel L-asparaginase was identified from forest soil metagenome and its characteristics were studied. The purified protein had a specific activity of 696IUmg-1 and optimum activity at pH 7 and 35°C. Enhanced enzyme activities were observed in the presence of Mg2+, Ca2+ and K+. The Km value, 2mM, and enzyme specificity constant 7.7mM-1s-1 indicated that the recombinant enzyme has good substrate affinity to L-asparagine compared with commercially-available Escherichia coli asparaginase. The IC50 value of 0.78µgml-1 (0.47IUml-1) was observed with HL60 cell line and 0.39µgml-1(0.23IUml-1) with MOLT-3 and MOLT-4 cell lines, which is better than that of commercially-available drugs. The soil metagenome derived L-asparaginase with enhanced activities could be a potential candidate to develop as a drug in Acute Lymphoblastic Leukemia (ALL) therapy.
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