Lipase From Rhizomucor miehei Immobilized on Magnetic Nanoparticles: Performance in Fatty Acid Ethyl Ester (FAEE) Optimized Production by the Taguchi Method

Abstract

In this communication, it was evaluated the production of fatty acid ethyl ester (FAAE) from the free fatty acids of babassu oil catalyzed by lipase from Rhizomucor miehei (RML) immobilized on magnetic nanoparticles (MNP) coated with 3-aminopropyltriethoxysilane (APTES), Fe 3 O 4 @APTES-RML or RML-MNP for short. MNPs were prepared by co-precipitation coated with 3-aminopropyltriethoxysilane and used as a support to immobilize RML (immobilization yield: 94.7 ± 1.0%; biocatalyst activity: 341.3 ± 1.2 U p−NPB /g), which were also activated with glutaraldehyde and then used to immobilize RML (immobilization yield: 91.9 ± 0.2%; biocatalyst activity: 199.6 ± 3.5 U p−NPB /g). RML-MNP was characterized by X-Ray Powder Diffraction (XRPD), Fourier Transform-Infrared (FTIR) spectroscopy and Scanning Electron Microscope (SEM), proving the incorporation and immobilization of RML on the APTES matrix. In addition, the immobilized biocatalyst presented at 60 • C a half-life 16-19 times greater than that of the soluble lipase in the pH range 5-10. RML and RML-MNP showed higher activity at pH 7; the immobilized enzyme was more active than the free enzyme in the pH range (5-10) analyzed. For the production of fatty acid ethyl ester, under optimal conditions [40 • C, 6 h, 1:1 (FFAs/alcohol)] determined by the Taguchi method, it was possible to obtain conversion of 81.7 ± 0.7% using 5% of RML-MNP.

Effect of Additional Amino Group to Improve the Performance of Immobilized Lipase From Aspergillus niger by Adsorption-Crosslinking Method

“…Cross-linking is an enzyme immobilization method without support and used only for bifunctional cross-linkers such as glutaraldehyde. Glutaraldehyde is a strong cross-linker, giving intermolecular cross-linking in any supports, coping the enzyme leakage, providing the stabilization of enzyme, and also functions as an activator to allow an enzyme-support covalent bond, and can be used to bind the enzymes to the matrix that has a free amino group (Broun, 1976;Barbosa et al, 2012;Barbosa et al, 2014;Moreira et al, 2020). Also, glutaraldehyde acts as a spacer arm to decrease inadmissible enzyme-functional group carrier surface interaction and minimize the steric hindrance (Yang et al, 2010).…”

Current Status and Future Perspectives of Supports and Protocols for Enzyme Immobilization

“…Physical and chemical interactions between supports and enzymes define the linkage of the immobilized biocatalyst [69][70][71][72][73]. The developed technologies and methods for this procedure still use one or more basic strategies: adsorption, encapsulation, covalent bonding, entrapment, and crosslinking.…”

Designing of Nanomaterials-Based Enzymatic Biosensors: Synthesis, Properties, and Applications

“…Despite its excellent performance potential, industrial applications were made impossible due to these undesirable characteristics [195,196]. In this scenario, immobilization techniques stand out as alternatives to overcome these limitations, since they can offer better stability, increased activity and selectivity, excellent resistance, improvements in product separation and purification, and the possibility of enzyme reuse, rendering processes increasingly efficient [192][193][194][195][196][197][198][199][200][201][202][203][204][205][206].…”

“…Enzymatic immobilization is based on the confinement of enzyme molecules on the surface of a reliable support that is different from that in which the substrate or products are present [202][203][204][205][206][207][208][209]. In contrast with their solubilized form, immobilized enzymes provide a large enzyme to substrate ratio, efficient digestion, and secure handling, in addition to showing more significant activity and the possibility of reuse for several cycles [197,[210][211][212][213]. The stability of free enzymes is mainly dictated by its intrinsic structure, while the stability of their immobilized counterparts is highly dependent on several other factors, as shown in Figure 4.…”