A flow-through strategy using supported ionic liquids for L-asparaginase purification

L-asparaginase (ASNase) is an amidohydrolase enzyme widely distributed in nature, e.g., microorganisms, plants and tissues of several animals. Nevertheless, microorganisms are the preferential source ASNase since they usually grow simple substrates culture conditions. However, a high level purity required by pharmaceutical industry, which applied downstream process may account for up to 80% total production costs. Silica-based supported ionic liquid-like phase (SSILLP) materials here proposed as alternative immobilization/capture or processing supports purification. SSILLP with different alkyl chain lengths at cation Cl− counterion were investigated purify flow-through like mode. Silica functionalized dimethylbutylpropylammonium chloride ([Si][N3114]Cl) was selected most promising material it displayed highest purification factor (1.65) specific activity (0.026 U mg−1) achieved. The operating conditions then optimized through Response Surface Methodology, using pH (range active) solid/liquid ratio (S/L ratio) factors, achieving maximum 3.36. Semi-continuous finally performed under (pH 3 S/L 15), enabling 5.15. This corresponds 3.12- 1.53-fold increase obtained compared initial screening batch assays These findings demonstrate that can act semi-continuous potential processing.