Development of a downstream process for purification and purity analysis of glutaminase free L-asparaginase using UPLC, DLS-ZP and DSC-TGA

Several bacterial strains were isolated from soil and water in the current work and screened for glutaminase-free L-asparaginase activity. The selected strain was subjected to the production of glutaminase-free L-asparaginase and thereby purification. Before purification, pH scouting was performed, and 8.6 pH was the optimum working pH to carry out purification. The enzyme was purified using Sephadex G-200 gel filtration chromatography and up to 4.55-folds and its purity percentage was determined using UPLC. The purity of L-asparaginase was determined to be 99.2%. The enzyme was characterized using UV-Visible Spectrophotometer, DLS, ZP, DSC, TGA. The loss of the enzyme was 65% after sublimation, indicating its purity compared to the standard drug. The results report a developed process for the purification of L-asparaginase and its characterization, which can prove instrumental in the purification process of naturally occurring enzymes because today, most developed processes deal with recombinant enzymes.

Automated summary

In this study, multiple bacterial strains were isolated from soil and water, screened for glutaminase-free L-asparaginase activity, and purified for further characterization. The optimal working pH for purification was found to be 8.6, resulting in a purity level of 99.2% for the L-asparaginase enzyme. This developed purification process and characterization could be instrumental for naturally occurring enzymes, as most current processes focus on recombinant enzymes.