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Ionic liquids (ILs) have evolved as a new type of non-aqueous solvents for biocatalysis, mainly due to their unique and tunable physical properties. A number of recent review papers have described a variety of enzymatic reactions conducted in IL solutions, on the other hand, to improve the enzyme's activity and stability in ILs; major methods being explored include the enzyme immobilization (on solid support, sol-gel, etc.), protic ionic liquids used as an additive process. The immobilization of the lipase from Burkholderia cepacia by the sol-gel technique using protic ionic liquids (PIL) as additives to protect against inactivation of the lipase due to release of alcohol and shrinkage of the gel during the sol-gel process was investigated in this study. The influence of various factors such as the length of the alkyl chain of protic ionic liquids (monoethanolamine-based) and a concentration range between 0.5 and 3.0% (w/v) were evaluated. The resulting hydrophobic matrices and immobilized lipases were characterised with regard to specific surface area, adsorption-desorption isotherms, pore volume (V-p) and size (d(p)) according to nitrogen adsorption and scanning electron microscopy (SEM), physico-chemical properties (thermogravimetric - TG, differential scanning calorimetry - DSC and Fourier transform infrared spectroscopy - FTIR) and the potential for ethyl ester and emulsifier production. The total activity yields (Y-a) for matrices of immobilized lipase employing protic ionic liquids as additives always resulted in higher values compared with the sample absent the protic ionic liquids, which represents 35-fold increase in recovery of enzymatic activity using the more hydrophobic protic ionic liquids. Compared with arrays of the immobilized biocatalyst without additive, in general, the immobilized biocatalyst in the presence of protic ionic liquids showed increased values of surface area (143-245 m(2) g(-1)) and pore size (19-38 angstrom). Immobilization with protic ionic liquids also favoured reduced mass loss according to TG curves (always less than 42.9%) when compared to the immobilized matrix without protic ionic liquids (45.1%), except for the sample containing 3.0% protic ionic liquids (46.5%), verified by thermogravimetric analysis. Ionic liquids containing a more hydrophobic alkyl group in the cationic moiety were beneficial for recovery of the activity of the immobilized lipase. The physico-chemical characterization confirmed the presence of the enzyme and its immobilized derivatives obtained in this study by identifying the presence of amino groups, and profiling enthalpy changes of mass loss. (c) 2013 Elsevier Inc. All rights reserved.

To create compatible interface for enzyme immobilization, the surface of multi-walled carbon nanotubes (MWCNTs) was functionalized using soft technique dielectric barrier discharge plasma (DBDP) for carboxylation and amination; followed by further amidation of carboxyl group with alkylamine. Successful functionalization and enzyme immobilization were structurally confirmed using spectroscopic analysis Fourier-Transform Infrared Spectroscopy (FTIR) and X-ray Photoelectron Spectroscopy (XPS). The immobilization of Candida rugosa lipase (CRL) on functionalized MWCNTs was evidenced by clearly viewing with Transmission Electron Microscopy (TEM) and Atomic Force Microscopy (AFM) imaging. CRL showed more Freundlich equilibrium behavior upon immobilization on annealed and octadecylamidated MWCNTs, which suggested a multilayer adsorption; while upon physical adsorption on aminated and carboxylated MWCNTs, CRL, to more extent, demonstrated a Langmuir equilibrium property, producing an enzyme monolayer. It was proven that DBDP-mediated surface-functionalization could create compatible microenvironments for enzyme immobilization, resulted in improved specific activity and thermostability. The immobilized CRL on octadecylamidated MWCNTs displayed excellent reusability and operation stability, indicating its potential for industrial application. (C) 2014 Elsevier B.V. All rights reserved.