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

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.