Jaya Kumar Arjun1, Balakrishna Pillai Aneesh1, Thulasi Kavitha1, Kumarapillai Harikrishnan2. 1. Environmental Biology Lab, Department of Biotechnology, Government of India, Rajiv Gandhi Centre for Biotechnology, Thycaud P.O. Poojappura, Thiruvananthapuram, Kerala, 695014, India. 2. Environmental Biology Lab, Department of Biotechnology, Government of India, Rajiv Gandhi Centre for Biotechnology, Thycaud P.O. Poojappura, Thiruvananthapuram, Kerala, 695014, India. harikrishnan@rgcb.res.in.
Abstract
OBJECTIVES: To screen soil metagenomic libraries for novel enzymes with enhanced activities. RESULTS: 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 696 IU mg-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, 2 mM, and enzyme specificity constant 7.7 mM-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 0.78 µg ml-1 (0.47 IU ml-1) was observed with HL60 cell line and 0.39 µg ml-1(0.23 IU ml-1) with MOLT-3 and MOLT-4 cell lines, which is better than that of commercially-available drugs. CONCLUSION: 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.
OBJECTIVES: To screen soil metagenomic libraries for novel enzymes with enhanced activities. RESULTS: 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 696 IU mg-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, 2 mM, and enzyme specificity constant 7.7 mM-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 0.78 µg ml-1 (0.47 IU ml-1) was observed with HL60 cell line and 0.39 µg ml-1(0.23 IU ml-1) with MOLT-3 and MOLT-4 cell lines, which is better than that of commercially-available drugs. CONCLUSION: 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.
Authors: Werner Alfinito Feio de Moura; Leonardo Schultz; Carlos Alexandre Breyer; Ana Laura Pires de Oliveira; Carlos Abrunhosa Tairum; Gabriella Costa Fernandes; Marcos Hikari Toyama; Adalberto Pessoa-Jr; Gisele Monteiro; Marcos Antonio de Oliveira Journal: Biotechnol Lett Date: 2020-07-07 Impact factor: 2.461