A successful process to extract lipolytic enzymes based on an aqueous biphasic system (ABS), which uses both ionic liquids (ILs) and a high charge-density inorganic salt (K₂CO₃), is proposed in this work. The activity of a model Thermomyces lanuginosuslipase (TlL) in some of the most common hydrophilic ILs, based on the 1-alkyl-3-methylimidazolium cation, combined with chloride, alkylsulfate, alkylsulfonate and acetate, was investigated. Several operating conditions influencing lipase activity and ABS formation were investigated. Parameters such as temperature, pH, deactivation kinetics and water content were evaluated in order to propose a viable extraction process. A deeper analysis in terms of enzyme deactivation kinetics was carried out, and the data were modelled through a series-type deactivation equation. ATR-FTIR studies aimed at identifying the TlL structure in selected ILs have also provided an insight into the enzyme deactivation behaviour.

An integrated process based on the concomitant role of non-ionic surfactants as cell disrupters and organic extractants of intracellular antioxidants is proposed for the first time in this work. The lytic effect of two common families of surfactants on the cell walls of the microalga Tetraselmis suecica has been initially evaluated. The antioxidant extraction based on aqueous two-phase systems (ATPS) was proposed using different potential salting out agents such as sodium inorganic and organic salts in aqueous solutions of the selected surfactants. The extraction efficiency was ascertained for the most representative biomolecules previously detected in this microalga: α-tocopherol, β-carotene and gallic acid. The viability of the process was checked in real lyophilized microalga samples, yielding higher antioxidant activity than that provided by an ultrasound-based conventional method.

One of the challenges in the field of biocatalysis is the search for efficient reaction media avoiding enzyme deactivation.