Industrial effluent as a substrate for glutaminase free l-asparaginase production from Pseudomonas plecoglossicida strain RS1; media optimization, enzyme purification and its characterization

Abstract

Glutaminase free L-asparaginase is a vital enzyme because of its anticancer potential. A potent bacterium isolated from a marine environment which produces glutaminase free L-asparaginase using M-9 medium with L-asparagine, was identified as Pseudomonas plecoglossicida RS1 by 16S rRNA gene sequencing. Statistical modeling was employed to optimize the medium using sugar cane industry effluent as the sole substrate for L-asparaginase production. The enzyme activity of L-asparaginase was higher with M-9 medium containing 0.8% effluent (3.25 ± 0.12 IU mL⁻¹) compared to M-9 medium containing 0.3% L-asparagine (0.73 ± 0.08 IU mL⁻¹). The apparent K_m and V_max of the purified L-asparaginase was 2.25 ± 0.61 mM and 8.9 ± 0.81 IU mL⁻¹ min⁻¹ respectively and the optimal activity of L-asparaginase was at pH 8.5 and 55 °C. This study highlights the use of industrial effluent as an alternate to L-asparagine for the production of L-asparaginase and how to improve the cost effectiveness of this enzyme.