A potential type-II L-asparaginase from marine isolate Bacillus australimaris NJB19: Statistical optimization, in silico analysis and structural modeling

L-asparaginase is a cardinal biotherapeutic drug for treating acute lymphoblastic leukemia, which is highly prevalent in children worldwide. In the current investigation, L-asparaginase producing marine bacterial isolate, Bacillus australimaris NJB19 (MG734654), was observed to be producing extracellular glutaminase free L-asparaginase (1327 +/- 0.4 IU mL(-1)). Production of L-asparaginasc was enhanced by the Box-Behnken design approach that enumerated the significant variables affecting the enzyme production. The optimum levels of the derived variables resulted in 2.8-fold higher levels of the enzyme production (37.93 +/- 1.06 IU mL(-1)). An 1146 bp L-asparaginase biosynthetic gene of Bacillus australimaris NJB19 was identified and cloned in E. call DH5a, fused with a histidine tag. The in silico analysis of the protein sequence revealed the presence of a signal peptide and classified it as a type II L-asparaginase. Toxic peptide prediction disclosed no toxin domain in the protein sequence, hence suggesting it as a non-toxic protein. The secondary structure analysis of the enzyme displayed a comparable percentage of alpha-helical and random coil structure, while 14.39% and 6.57% of amino acid residues were composed of extended strands and beta-turns, respectively. The functional sites in the three-dimensional structural model of the protein were predicted and interestingly had a few less conserved residues. Bacillus australimaris NJB19 identified in this study produces type-II L-asparaginase, known for its high affinity for asparagine and effectiveness against leukemic cells. Hence, these observations indicate the Lasparaginase, thus obtained, as a potentially significant and novel therapeutic drug. (C) 2021 Elsevier B.V. All rights reserved.