Additives Enhancing the Catalytic Properties of Lipase from Burkholderia cepacia Immobilized on Mixed-Function-Grafted Mesoporous Silica Gel

Abstract

Effects of various additives on the lipase from Burkholderia cepacia (BcL) immobilized on mixed-function-grafted mesoporous silica gel support by hydrophobic adsorption and covalent attachment were investigated. Catalytic properties of the immobilized biocatalysts were characterized in kinetic resolution of racemic 1-phenylethanol (rac-1a) and 1-(thiophen-2-yl)ethan-1-ol (rac-1b). Screening of more than 40 additives showed significantly enhanced productivity of immobilized BcL with several additives such as PEGs, oleic acid and polyvinyl alcohol. Effects of substrate concentration and temperature between 0-100 °C on kinetic resolution of rac-1a were studied with the best adsorbed BcLs containing PEG 20 k or PVA 18-88 additives in continuous-flow packed-bed reactor. The optimum temperature of lipase activity for BcL co-immobilized with PEG 20k found at around 30 °C determined in the continuous-flow system increased remarkably to around 80 °C for BcL co-immobilized with PVA 18-88.

Selection of Lipases for the Synthesis of Biodiesel from Jatropha Oil and the Potential of Microwave Irradiation to Enhance the Reaction Rate

“…These additives have been used to protect the enzyme from external denaturing agents by providing additional sites for hydrogen bonding with the enzyme surface that decreases dehydration and distortion of its three-dimensional structure [39]. However, these additives can interact with nucleophilic groups in the enzyme structure (e.g., –NH 2 and –OH) that influence the orientation of enzyme molecules on the support surface [40, 41]. Similar results were reported in previous studies carried out in our lab concerning the immobilization of Thermomyces lanuginosus lipase from different commercial formulations supplied by Novozymes (Lipolase® and Lipex® 100 L) by different protocols on activated organic supports (glyoxyl-agarose and epoxy-chitosan/alginate beads) [23, 24].…”

Strategies for the one-step immobilization–purification of enzymes as industrial biocatalysts

“…The problem is the necessity of having at least a primary amino group near the area involved in the immobilization., in In a recent paper it has been shown that this may (Figure 17) (Rueda et al, 2014). In other papers, amino-phenyl silicates were used to immobilize the lipases from C. antarctica (form B) (Boros et al, 2013) and Burkholderia cepacia (Abaházi et al, 2014). After adsorption, the immobilized enzymes were treated with glycerol diglycidyl ether to have some enzyme-support crosslinking.…”

“…Immobilization was suggested to proceed via interfacial activation (activity improved after immobilization), and the final covalent attachments improve enzyme stability (Abaházi et al, 2014, Boros et al, 2013 This kind of heterofunctional hydrophobic supports may greatly improve the prospects of using interfacial activation of lipases as a tool to prepare biocatalysts useful under any experimental conditions, using the hydrophobic groups to have the a selective adsorption, and the other groups to transform this in an irreversible immobilization. The change of reversible to irreversible immobilization means that the support cannot be reused, therefore the strategy is only recommended when the improvement in enzyme stability or the prevention of enzyme release to the reaction medium are really relevant.…”

Cross‐linked enzyme aggregates of recombinant Candida antarctica lipase B for the efficient synthesis of olvanil, a nonpungent capsaicin analogue

“…The tailor‐made enzymes obtained by bioimprinting are limited to nonaqueous dry organic solvents, as the induced catalytic active site created by the bioimprint molecule is lost in contact to aqueous solutions . To overcome this limitation, the induced complex can be locked by immobilization on aminated or hydrophobic supports or trapped by cross‐linking upon precipitation . This technique not only allows the enhancement of the activity and selectivity of immobilized enzymes, but also the specificity and thermal stability of lipases, and can be strategically combined with an induced interfacial activation to enhance the catalytic properties of lipases.…”