Lipolytic Enzymes with Improved Activity and Selectivity upon Adsorption on Polymeric Nanoparticles

Abstract

Nanostructured polystyrene (PS) and polymethylmethacrylate (PMMA) were used as carriers for the preparation of bioconjugates with lipolytic enzymes, such as Candida rugosa lipase (CRL) and Pseudomonas cepacia lipase (PCL). Simple addition of the lipase solution to the polymeric nanoparticles under protein-friendly conditions (pH 7.6) led to the formation of polymer-enzyme bioconjugates. Energy filtered-transmission electron microscopy (EF-TEM) performed on immuno-gold labeled samples revealed that the enzyme preferentially binds to the polymer nanoparticles and that the binding does not affect the nanostructured features of the carriers. The studies performed on the activity of the bioconjugates pointed out that the lipases adsorbed onto polymeric nanoparticles show an improved performance in terms of activity and selectivity with respect to those shown by lipases adsorbed on the same non-nanostructured carriers. The residual activities of CRL and PCL immobilized on nanostructured PMMA and PS reached 60% and 74%, respectively. Moreover, we found that enantioselectivity and pH and thermal stability increase upon immobilization. These results highlight the fact that new protein conformers with improved enantioselectivity stabilized after adsorption on nanoparticles are obtained. On the basis of the chemical structures of the selected polymers and the slopes of the adsorption isotherms, a hydrophobic binding model for lipase/nanostructured polymers is suggested.

Immobilization of Lipases onto Magnetic Fe3O4 Nanoparticles for Application in Biodiesel Production

“…Compared to studies by other researchers, the immobilized lipases were more stable during reuse and storage. [6,8,15,16] Previous reports showed that lipase from Paeudomonas cepacia has the greatest methanol resistance among different lipases; consequently, the Paeudomonas cepacia lipase (lipase L3) has often been used in enzymatic transesterification reactions. [17,18] In this study, the immobilized lipase L3 was investigated for its activity in biodiesel production.…”

Design of novel nano-carriers for multi-enzyme co-localization

“…Enzymes can be attached onto polymeric nanoparticles via physical adsorption, covalent bonding, cross-linking or a combination thereof (Akgol et al, 2009;Daubresse et al, 1994;Matsuno and Ishihara, 2009;Palocci et al, 2007;Watanabe and Ishihara, 2004). Various polymeric materials and immobilization strategies have been designed for attaching enzymes.…”

Hybrid Systems Biomolecule-Polymeric Nanoparticle: Synthesis, Properties and Biotechnological Applications

“…Also, many enzymes, e.g., lipases, improve their activity upon the adsorption onto nanostructured carrier materials [106].…”

“…Enzymes such as Candida rugosa lipase (CRL) and Pseudomonas cepacia lipase (PCL) have been adsorbed on nanostructured polystyrene (PS) and polymethylmethacrylate (PMMA) by simple addition of the lipase solution to the polymeric nanoparticles under protein-friendly conditions (pH 7.6). Adsorption leads to improved performance in terms of activity and selectivity with respect to that shown by lipases adsorbed on the same nonnanostructured carriers, as well as increased enantioselectivity and pH and thermal stability [106]. Karajanagi et al have also examined the structure and function of two enzymes, R-chymotrypsin (CT) and soybean peroxidase (SBP), adsorbed onto single-walled carbon nanotubes (SWNTs).…”

“…In this regard, nanostructured materials will provide the upper limits in terms of balancing the contradictory issues including surface area, mass transfer resistance, and effective enzyme loading [119]. The reported works in this area have revealed the great potential for the use of nanoporous [120], nanofibrous [119], and nanoparticle [106] materials as new classes of carriers for biocatalysts. The effective enzyme loading on nanomaterials can be very high (for example, it can reach over 10 wt% with particles smaller than 100 nm [122], and a large surface area per unit mass is also provided to facilitate reaction kinetics).…”