Optimization of aqueous two-phase micellar system for partial purification of L-asparaginase from Penicillium sp. grown in wheat bran as agro-industrial residue

Abstract

L-asparaginase has been used in the remission of malignant neoplasms such as acute lymphoblastic leukemia. The search for new sources of this enzyme has become attractive for therapeutics. Traditional methods for biomolecule purification involve several steps. A two-phase system may be a good strategy to anticipate one of these stages. This study aimed to produce and purify a fungal L-asparaginase through an aqueous two-phase micellar system (ATPMS) using Triton X-114. The fungus Penicillium sp.-encoded 2DSST1 was isolated from Cerrado soil. Plackett-Burman design followed by a 2 4 full factorial design was used to determine the best conditions to produce L-asparaginase. The evaluated variables were L-asparagine, L-proline, wheat bran, potato dextrose broth, ammonium sulfate, yeast extract, sucrose and glucose concentrations, incubation temperature, incubation period, and initial pH of the culture medium. L-asparaginase quantification was valued by the formation of β-aspartyl hydroxamate. The significant positive variables, L-asparagine, L-proline, potato dextrose broth, and sucrose concentrations, were evaluated at 2 levels (+ 1 and − 1) with triplicate of the central point. After 34 runs, maximum activity (2.33 IU/mL) was achieved at the factorial design central point. A central composite design was performed in ATPMS at two levels (+ 1 and − 1) varying Triton X-114 concentration (w/v), separation phase temperature, and crude extract concentration (w/v). The Lasparaginase partition coefficient (K) was considered the experimental design response. Out of the 16 systems that were examined, the most promising presented a purification factor of 1.4 and a yield of 100%.

Production, characterization and techno-economic evaluation of Aspergillus fusant l-asparaginase

“…Molecular weight of l -ASNase varies between 33 and160 kDa (Cardoso et al 2020 ) which accommodates the 116.4 kDa of Fusant-06 l -ASNase in this study. An l -ASNase with molecular weight of 115 kDa from Aspergillus oryzae CCT 3940 has been reported by Dias et al ( 2016 ).…”

Microbial L-asparaginase for Application in Acrylamide Mitigation from Food: Current Research Status and Future Perspectives

“…The aqueous two-phase system (ATPS) is a liquid–liquid fractionation method used as a pre-purification method. Cardoso et al (2020) purified L-asparaginase production of the fungus Penicillium sp.–encoded 2DSST1 through ATPMS using Triton X-114, and obtained a purification factor of 1.4 and a yield of 100% [ 62 ]. Nickel affinity chromatography is also used to purify the recombinant L-asparaginase, with a polyhistidine tag on either terminus [ 20 ].…”

Potential of Anti-Cancer Activity of Secondary Metabolic Products from Marine Fungi

“…[63] The application of L-asparaginase is presented in Table 7. At a concentration of 125 µg/mL, L-Asparaginase showed high activity against lymphoma cancer cells (U937), with an IC 50 value of 500 µg/mL of β-cyclodextrin-asparaginase nanobiocomposite [70]. In the anti-cancer studies of Baskar et al [6], the L-Asparaginase is incorporated into the nano biocomposites which have been synthesized using β-cyclodextrin and chitosan to investigate the anti-cancer activity against prostate cancer cell lines and lymphoma cancer cells.…”