Influence of Glutaraldehyde Cross-Linking Modes on the Recyclability of Immobilized Lipase B from Candida antarctica for Transesterification of Soy Bean Oil

Abstract

Lipase B from Candida antarctica (CAL-B) is largely employed as a biocatalyst for hydrolysis, esterification, and transesterification reactions. CAL-B is a good model enzyme to study factors affecting the enzymatic structure, activity and/or stability after an immobilization process. In this study, we analyzed the immobilization of CAL-B enzyme on different magnetic nanoparticles, synthesized by the coprecipitation method inside inverse micelles made of zwitterionic surfactants, with distinct carbon chain length: 4 (ImS4), 10 (ImS10) and 18 (ImS18) carbons. Magnetic nanoparticles ImS4 and ImS10 were shown to cross-link to CAL-B enzyme via a Michael-type addition, whereas particles with ImS18 were bond via pyridine formation after glutaraldehyde cross-coupling. Interestingly, the Michael-type cross-linking generated less stable immobilized CAL-B, revealing the influence of a cross-linking mode on the resulting biocatalyst behavior. Curiously, a direct correlation between nanoparticle agglomerate sizes and CAL-B enzyme reuse stability was observed. Moreover, free CAL-B enzyme was not able to catalyze transesterification due to the high methanol concentration; however, the immobilized CAL-B enzyme reached yields from 79.7 to 90% at the same conditions. In addition, the transesterification of lipids isolated from oleaginous yeasts achieved 89% yield, which confirmed the potential of immobilized CAL-B enzyme in microbial production of biodiesel.

Novozym 435: the “perfect” lipase immobilized biocatalyst?

“…42 The heterogeneity of the natural substrate [43][44][45][46][47][48] has converted lipases in enzymes with a very broad specificity, accepting substrates very different from glycerides (even amides). Thus, lipases are used in vitro to catalyze reactions different from those of the natural hydrolase function, [49][50][51][52][53][54] such as esterification, [55][56][57][58][59][60][61] acidolysis, [62][63][64][65][66][67] interesterificaton, [68][69][70][71] transesterification, [72][73][74][75][76] aminolysis, [77][78][79][80][81] perhydrolysis, 82,83 etc., together with a collection of the so-called promiscuous reactions. [84][85][86][87][88]...…”

Lecitase ultra: A phospholipase with great potential in biocatalysis

“…Among the most used enzymes, lipases stand out [42,43]. The natural function of lipases is the hydrolysis of triglycerides to free fatty acids and glycerol [44]; however, in vitro lipases have been used in a wide variety of reactions, some related to oil modifications (esterifications [45][46][47][48][49][50][51], transesterifications [52][53][54][55][56], acidolysis [57][58][59][60][61][62], interesterifcations [63][64][65][66]), some far from these substrates (resolution of racemic mixtures, regio or enantio selective reactions) and some have even exhibited a diversity of the so-called promiscuous activities far from their natural function [67][68][69][70][71][72][73][74][75].…”

Enzymatic Reetherification in the Production of Butterfat Substitutes

“…Переэтерифицированный жир насосом подают на полировочные фильтры, а затем на окончательный охладитель. Охлажденный переэтерифицированный жир перекачивают в бак для накопления, затем направляют на дезодорацию [11].…”