Enzymatic production of wax esters by esterification using lipase immobilized via physical adsorption on functionalized rice husk silica as biocatalyst.

The present study consists of developing an enzymatic process for the production of wax esters (lauryl stearate and cetyl stearate) by esterification in a heptane medium. Lipase from Thermomyces lanuginosus (TLL) immobilized via interfacial activation on silica particles from rice husks functionalized with triethoxy(octyl)silane (TLL-Octyl-SiO2 ) was used as biocatalyst. Maximum immobilized protein loading of around 22 mg g-1 (that corresponds to an immobilization yield of ≈55%) of support was observed using an initial protein loading of 40 mg g-1 of Octyl-SiO2 . Its hydrolytic activity (olive oil emulsion hydrolysis) was of 620 U g-1 of biocatalyst. The effect of certain factors on the cetyl estearate production was evaluated using a central composite rotatable design (CCDR). Under optimal conditions (64°C, 21% of mass of biocatalyst per volume of reaction mixture, 170 rpm, and stoichiometric acid:alcohol molar ratio 1 mol L-1 of each reactant), maximum acid conversion percentage of 91% was observed after 60 min of reaction. Lauryl stearate was also produced under such conditions, and an acid conversion of 93% after 60 min of reaction was also achieved. Free lipase exhibited acid conversion of only 15%-20% for both reaction mixtures. After nine successive esterification batches, TLL-Octyl-SiO2 retained 85%-90% of its original activity. These results show the promising use of the prepared biocatalyst in wax esters production due to its high catalytic activity and reusability.

Decyl esters production from soybean-based oils catalyzed by lipase immobilized on differently functionalized rice husk silica and their characterization as potential biolubricants.

This study aimed the enzymatic decyl esters production by hydroesterification, a two-step process consisting of hydrolysis of refined soybean (RSBO) or used soybean cooking (USCO) oils to produce free fatty acids (FFA) and further esterification of purified FFA. Using free lipase from Candida rugosa (CRL), about 98% hydrolyses for both oils have been observed after 180 min of reaction using a CRL loading of 50 U g-1 of reaction mixture, 40 °C, and a mechanical stirring of 1500 rpm. FFA esterification with decanol in solvent-free systems was performed using lipase from Thermomyces lanuginosus (TLL) immobilized by physical adsorption on silica particles extracted from rice husk, an agricultural waste. For such purpose, non-functionalized (SiO2) or functionalized rice husk silica bearing octyl (Octyl-SiO2) or phenyl (Phe-SiO2) groups have been used as immobilization supports. Protein amounts between 22 and 28 mg g-1 of support were observed. When used in the esterification, they enabled a FFA conversion of 81.3-87.6% after 90-300 min of reaction. Lipozyme TL IM, a commercial immobilized TLL, exhibited similar performance compared to TLL-Octyl-SiO2 (FFA conversion ≈90% after 90-120 min of reaction). However, high operational stability after fifteen successive esterification batches was observed only for TLL immobilized on Octyl-SiO2 (activity retention of ≈90% using both FFA sources). The produced decyl esters presented good characteristics as potential biolubricants according to standard methods (ASTM) and thermal analysis.

Improved immobilization of lipase from Thermomyces lanuginosus on a new chitosan-based heterofunctional support: Mixed ion exchange plus hydrophobic interactions.

In this study, a new mixed heterofunctional support (Chit-GA-Gly) has been prepared by sequential activation of chitosan hydrogel (Chit) with glutaraldehyde (GA) and further functionalization with glycine (Gly). The immobilization of the lipase from Thermomyces lanuginosus (TLL) on this support was compared with that on GA-activated Chit hydrogel (Chit-GA). The supports have been characterized by FT-IR, zeta potential and TG analyses. A similar maximum lipase loading of 53-55 mg per gram of support has been obtained for both supports. Both biocatalysts retained ≈40% of their initial activity after 48 h of incubation at 50 °C in heptane, toluene or iso-octane. The immobilization of TLL on Chit-GA proceeded via preferential covalent attachment (95%) and a combined ion exchange (cationic and anionic) and hydrophobic adsorption was observed using Chit-GA-Gly. TLL immobilized on Chit-GA-Gly was ≈4-times more active than when immobilized on Chit-GA in both olive oil emulsion hydrolysis and alkyl palmitate synthesis via esterification. Isoamyl palmitate synthesis in iso-octane at 50 °C using this new biocatalyst gave a maximum acid conversion of 85% after 90 min of reaction. After nine consecutive esterification batches, the biocatalyst retained around 40% of its initial activity.