Thermally Stable and Reusable Silica and Nano-Fructosome Encapsulated CalB Enzyme Particles for Rapid Enzymatic Hydrolysis and Acylation

Abstract

This study reports the preparation of silica-coated and nano-fructosome encapsulated Candida antarctica lipase B particles (CalB@NF@SiO2) and a demonstration of their enzymatic hydrolysis and acylation. CalB@NF@SiO2 particles were prepared as a function of TEOS concentration (3–100 mM). Their mean particle size was 185 nm by TEM. Enzymatic hydrolysis was performed to compare catalytic efficiencies of CalB@NF and CalB@NF@SiO2. The catalytic constants (Km, Vmax, and Kcat) of CalB@NF and CalB@NF@SiO2 were calculated using the Michaelis–Menten equation and Lineweaver–Burk plot. Optimal stability of CalB@NF@SiO2 was found at pH 8 and a temperature of 35 °C. Moreover, CalB@NF@SiO2 particles were reused for seven cycles to evaluate their reusability. In addition, enzymatic synthesis of benzyl benzoate was demonstrated via an acylation reaction with benzoic anhydride. The efficiency of CalB@NF@SiO2 for converting benzoic anhydride to benzyl benzoate by the acylation reaction was 97%, indicating that benzoic anhydride was almost completely converted to benzyl benzoate. Consequently, CalB@NF@SiO2 particles are better than CalB@NF particles for enzymatic synthesis. In addition, they are reusable with high stability at optimal pH and temperature.

Enzymatic Degradation Behavior of Self-Degradable Lipase-Embedded Aliphatic and Aromatic Polyesters and Their Blends

“…These values were as expected, justifying the selection of this pH value for the degradation experiments due to the higher enzymatic activity at pH 8. 27 Finally, the Pluronic batch presented slightly lower U values, probably due to the lower protein content. Similar observations are reported in the literature.…”

“…Regarding the enzymatic activity, in all cases, CalB exhibited higher activity at pH 8 (10–46% higher, up to ∼91% in one of the batches); mean values were 0.2743 μmol·s –1 ·mg –1 at pH 8, and 0.2084 μmol·s –1 ·mg –1 at pH 7. These values were as expected, justifying the selection of this pH value for the degradation experiments due to the higher enzymatic activity at pH 8 . Finally, the Pluronic batch presented slightly lower U values, probably due to the lower protein content.…”

“…Candida antarctica lipase B (CalB) is a widely employed enzyme for polymerization reactions (esterification) in organic solvents and depolymerization (hydrolysis of esters) in aqueous media . Previous studies have shown the best conditions for this lipase in terms of enzymatic activity, which is directly related to depolymerization and polymeric degradation: the optimum and most stable pH is in the range of 8–9 and temperature in the range of 35–40 °C. , However, it remains stable from pH 3.5 to 9.5 in polar organic solvents and at temperatures as high as 150 °C . There are several studies in the literature regarding the enzymatic degradation of PLA, PBAT, and PBS, by externally adding CalB, which reported a wide range of results.…”

Ultrathin silica-tiling on living cells for chemobiotic catalysis

“…For proof-of-concept, we encapsulated Candida Antarctica Lipase B (CALB) inside 2D-SiNTs (designated as CALB/2D-SiNTs), characterized by Fourier transform infrared spectroscopy (FTIR) and Raman spectroscopy (Supplementary Figs. 23 – 25 ) 46 . After assembling CALB/2D-SiNTs on yeast (designated as Yeast@CALB/2D-SiNTs), we planned a two-step reaction sequence: yeast-mediated reduction of aldehyde to benzyl alcohol followed by CALB-mediated trans-esterification in one-pot (Fig.…”

Enhanced catalytic performance of penicillin G acylase by covalent immobilization onto functionally-modified magnetic Ni0.4Cu0.5Zn0.1Fe2O4 nanoparticles

“…Specifically, we focused on the use of magnetic Ni 0.4 Cu 0.5 Zn 0.1 Fe 2 O 4 nanoparticles, which were prepared using an advanced, convenient, simple, and rapid combustion process. To enhance the nanoparticles' functionality, a SiO 2 functional layer was applied to their surface using sodium silicate [63,64]. The modification process with sodium silicate introduced a substantial amount of Si-OH groups on the surface of the nanoparticles.…”