Abstract
AbstractBACKGROUNDl‐Asparaginase (ASNase) is an important biopharmaceutical for the treatment of acute lymphoblastic leukemia (ALL); however, with some restrictions due to its high manufacturing costs. Aqueous biphasic systems (ABS) have been suggested as more economical platforms for the separation/purification of proteins, but a full understanding of the mechanisms behind the ASNase partition is still a major challenge. Polymer/salt‐based ABS with different driving‐forces (salting‐out and hydrophilicity/hydrophobicity effects) were herein applied to control the partition of commercial ASNase.RESULTSThe main results showed the ASNase partition to the salt‐ or polymer‐rich phase depending on the ABS studied, with extraction efficiencies higher than 95%. For systems composed of inorganic salts, the ASNase partition was controlled by the polyethylene glycol (PEG) molecular weight used. Cholinium‐salts‐based ABS were able to promote a preferential ASNase partition to the polymer‐rich phase using PEG‐600 and to the salt‐rich phase using a more hydrophobic polypropylene glycol (PPG)‐400 polymer. It was possible to select the ABS composed of PEG‐2000 + potassium phosphate buffer as the most efficient to separate the ASNase from the main contaminant proteins (purification factor = 2.4 ± 0.2), while it was able to maintain the enzyme activity for posterior application as part of a therapeutic.CONCLUSIONPolymer/salt ABS can be used to control the partition of ASNase and adjust its purification yields, demonstrating the ABS potential as more economic platform for the selective recovery of therapeutic enzymes from complex broths. © 2019 Society of Chemical Industry
Highlights
Violacein is a natural purple-blue hydrophobic pigment[1] with interesting biological activities such as antitumoral, antiparasitic, antifungal, antiviral, antiprotozoal, antioxidant, antiulcerogenic and immunomodulatory.[2]
This is probably due to the low ability of hydrophilic ionic liquids (ILs) to disrupt cell membranes, considering their low affinity to interact with the phospholipids composing the membranes, due to their hydrophobic nature.[42,48]
The ability to induce these changes in cell permeability and/or to solubilize violacein is dependent on the surfactant type, which is demonstrated by the intensity in the lysate supernatant
Summary
Violacein is a natural purple-blue hydrophobic pigment[1] with interesting biological activities such as antitumoral, antiparasitic, antifungal, antiviral, antiprotozoal, antioxidant, antiulcerogenic and immunomodulatory.[2]. The available methods for intracellular compound release are normally divided into two main groups, mechanical and non-mechanical techniques.[5] Mechanical techniques (bead mill, homogenization and ultrasonic treatment) are easy to scale up. They present a non-selective character and can negatively affect the biological activity of the target compounds and the downstream process owing to the finer cell debris resulting from the high degree of disruption. Violacein is a natural purple-blue hydrophobic pigment with interesting bioactivity whose expression in genetically modified Yarrowia lipolytica production was successfully achieved
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