Optimization, purification, and characterization of L-asparaginase from Aspergillus quadrilineatus

Abstract

One remedy used to treat lymphoblastic cancer is L-asparaginase. The ability of a new strain to produce extracellular L-asparaginase was investigated in this work. The study examined the production of L-asparaginase utilising MCDB, which exhibited the highest level of L-asparaginase activity. The optimisation, synthesis, purification, and characterisation of the Aspergillus quadrilineatus derived L-Asparaginase enzyme are the main objectives of this investigation. After the crude enzyme was removed, it was precipitated with ammonium sulphate, filtered using a Sephadex column, and then further purified using ion exchange chromatography to create L-asparaginase. The enzyme’s reaction to different temperatures, pH values, substrate concentrations, incubation times, and SDS-PAGE analyses was evaluated in order to characterise it. To improve the synthesis of L-asparaginase, physical parameters like pH and nutritional parameters like carbon, nitrogen, ionic salts, amino acids, phosphate groups, surfactants, plant oil, and vitamins, as well as their corresponding concentrations, were optimised. The results of the study showed that 37°C and a pH of 8.0 were the most conducive to the synthesis of enzymes. The most effective sources of carbon and nitrogen were L-asparagine (0.5%) and maltose (0.5%), respectively. Vitamins (1%), sodium acetate (1%), citric acid (1%), tryptophan (1%), and sodium dihydrogen phosphate (1%), were found abundant in the yield. With the medium composition described in the paper, continuous fermentation can produce L-asparaginase on a large scale. Protein profiling was conducted using SDS-PAGE on the partly purified enzyme, resulting in a predicted molecular weight of the protein as 47KDa.