Production, Purification, Characterization And Anti-Leukemic Activity Of L-Asparaginase From Streptomyces Spp Aia 1 Section: Microbial And Enzyme Technology

Abstract

Future therapeutic applications of L asparaginase in the treatment of leukemia and lymphoid system cancers are expected to cause the demand for the enzyme to increase several times over. It is therefore imperative to find appropriate sources of production. The novel L asparaginase producer (Streptomyces sp AIA 1) produced in this study offers a proof of concept for its production. It was isolated and then optimized by a solid state fermentation process employing a low-cost, straightforward medium made of soybean meal and rice bran. Examined were the effects of seven distinct independent parameters on the synthesis of L-asparaginase in solid state fermentation. Using the Plackett–Burman design, the effects of seven distinct independent parameters were investigated with regard to L-asparaginase synthesis under solid state fermentation conditions. Ammonium sulfate, FeSO4.7H2O, and soybean meal were the main variables influencing the synthesis of L-asparaginase. Thus, the ideal values of these variables were ascertained by the use of central composite design. Ammonium sulfate was used to purify L-asparaginase, and the final purification fold was 39.87,this was achieved by Sephadex G25 column chromatography and DEAE cellulose 52 ion exchange chromatography. The SDS-PAGE approach yielded a monomeric molecular weight of 30 kDa for the purified L-asparaginase. When L-asparaginase was tested in vitro on the leukemic cancer cell line K562 and the normal cell line PBMC, it was discovered to exhibit potent anti-proliferative properties. The outcomes demonstrated that the K562 cell lines (IC50 = 17.63) was subjected to the greatest cytotoxic effect of L-asparaginase.