Ionic liquid-based aqueous biphasic system for lipase extraction

Abstract

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 (K 2 CO 3 ), is proposed in this work. The activity of a model Thermomyces lanuginosus lipase (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.

Ionic liquids for the concomitant use in extremophiles lysis and extremozymes extraction

“…The value obtained reached up to 96%, which is indicative of the high affinity of the selected ionic liquid for the extremozyme under study. This behaviour is coincident with the results reported for other mesophilic enzymes such as those produced by Candida antarctica A or Thermomyces lanuginosus (Deive et al, 2011a(Deive et al, , 2012c. In summary, the proposed integrated strategy combining the lytic and extraction potential of the selected ionic liquid has been demonstrated to be suitable for extremozymes recovery.…”

“…This separation technique, named aqueous biphasic systems (ABS) is thus the result of a complex competition between ionic liquids and salts for the water molecules. Different advantages have justified their application for the extraction of a portfolio of compounds with industrial interest, such as their mild operating conditions, short segregation time, or their low energy requirements (Freire et al, 2010) Among the different uses, the extraction of biomolecules such as lipases has been highlighted in different research works (Deive et al, 2011a;Ventura et al, 2012).…”

Evaluating Self-buffering Ionic Liquids for Biotechnological Applications

“…13 In bioseparations water miscible-ILs hold great potential as a feasible alternative for phase forming agents in aqueous biphasic systems (ABS) to replace conventional polymer and/or salt-rich phases, 14 for the purification of various enzymes including alcohol dehydrogenases, 15 lysozyme, 16 cyclodextrin glycosyltransferase, 17 peroxidases, 18 and lipases. [19][20][21][22] The need to maintain a stable pH during an enzyme-catalysed reaction is well established. [23][24] Therefore, most applications of ILs in biotechnological field involve the addition of buffers to stabilize hydrogen ion concentrations during enzymatic reaction.…”

Extraction and stability of bovine serum albumin (BSA) using cholinium-based Good's buffers ionic liquids

“…IL-based ABS have been used for the extraction of proteins and enzymes, while demonstrating to provide higher extraction efficiencies than the traditional PEG-based ABS [17][18][19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34]. Alkylimidazolium-based ILs combined with phosphate-based salts were the most widely investigated phase-forming components of ABS [15].…”