Immobilization of cellulase on magnetic nanoparticles for rice bran oil extraction in a magnetic fluidized bed

This paper presents a method for extracting rice bran oil using magnetic immobilized cellulase (MIC) in a magnetic fluidized bed (MFB). Cellulase was immobilized on Fe3O4/SiO x -g-P (glycydylmethacrylate) with an average grain size of 120 nm. The rice bran was hydrolyzed using MIC combined with magnetic immobilized alkaline protease to extract rice bran oil. Under intermittent conditions, the MIC concentration was 1.6 mg/g, the liquid to material ratio was 4:1, the enzymatic hydrolysis time was 150 min, and the oil yield was as high as 85.6 ± 1.20% at 55 °C. The fluid in the MFB had a magnetic field strength of 0.022 T, a flow velocity of 0.005 m/s, and an oil extraction rate of 90.3%. This provides a theoretical basis for the extraction of rice bran oil using the subsequent MFB hydroenzyme method.

IMMOBILIZATION OF CELLULASES ON CHITOSAN: APPLICATION FOR SUGARCANE BAGASSE HYDROLYSIS

In this work, a commercial cellulolytic cocktail was immobilized on glutaraldehyde activated chitosan gel. The chitosan concentration in the gel preparation, pH, immobilization time and enzymatic loading were evaluated. Immobilized cellulases showed better hydrolysis performance when an enzyme loading of 134 mg protein/g carrier was used for immobilization at pH 9.0 for 30 minutes. Hydrolysates with a glucose content of 13.43 and 10.35 g/L were obtained when Avicel and pretreated sugarcane bagasse were used as substrate, respectively. Immobilized cellulase lost 60% of its hydrolysis performance after 8 cycles using Avicel, and 75% after 6 cycles for sugarcane bagasse. The hydrolysis performance associated with the reuse of the immobilized cellulases indicates that an improvement in the immobilization of cellulases, coupled with an improvement in the pretreatment of lignocellulosic biomass, will allow the development of a continuous hydrolysis system with the enzyme retained in the reactor.

Covalent Immobilization of Cellulase onto Amino and Graphene Oxide Functionalized Magnetic Fe2O3/Fe3O4@SiO2 Nanocomposites

Magnetic Fe2O3/Fe3O4@SiO2 nanocomposites were prepared via the citric-alcohol solution combustion process. The obtained nanocomposites were characterized with SEM, XRD, VSM, TEM, EDS, HRTEM, and FTIR techniques. The results revealed that the magnetic Fe2O3/Fe3O4@SiO2 nanocomposites were successfully obtained with the average grain size of 87 nm and the saturation magnetization of 36 emu/g. After the surface of magnetic Fe2O3/Fe3O4@SiO2 nanocomposites was functionalized by amino group, the amino-functionalized Fe2O3/Fe3O4@SiO2-NH2 nanocomposites were loaded onto graphene oxide based on Mitsunobu reaction. Subsequently, the cellulase was immobilized onto Fe2O3/Fe3O4@SiO2-NH-GO nanocomposites by a glutaraldehyde-mediated Schiff base reaction. The immobilization conditions were optimized by adjusting the pH, temperature, and cellulase dose. The results revealed that optimized immobilization conditions were determined to be temperature of 50 °C, pH of 5, and cellulase solution of 0.1 mL. 97.3% cellulase were successfully immobilized under the optimal conditions. The catalytic performances of the immobilized cellulase were also evaluated. The maximum activity was achieved at pH 4, and 50 °C with cellulase solution of 0.4 mL.

A Method of Improving Physical Adsorption of Cellulase On Mesoporous Zr-Based MOF Surface: Enhanced Stability, Recyclability, And Catalytic Efficiency

In this paper, we aimed at developing immobilized cellulase biocatalyst by enhancing the anchor of cellulase on support surface. A mesoporous Zr-based MOF was first synthesized by biomineralization method using dextran as template. The resultant PVP-cellulase@CD-UIO-66-Zr exhibited a high loading capacity of 265 mg g− 1 support. The physical adsorption of cellulase on CD-UIO-66-Zr could be further enhanced by the capping of cellulase with PVP. About 83% of the activity of PVP-cellulase@CD-UIO-66-Zr could be retained after six cycles, and its equilibrium leakage ratio was 36% during thirty days’ leaching test. It was noting that about 80% of activity of immobilized PVP-cellulase@CD-UIO-66-Zr could be retained after incubation at 80°C for 1 hour. The immobilized cellulase exhibited higher pH stability, thermostability, storage stability and catalytic efficiency than free one.

A two-step method for the synthesis of magnetic immobilized cellulase with outstanding thermal stability and reusability

In order to make better use of cellulase, a simple and fast method using electrostatic attraction and silica embedment was proposed for enzyme immobilization. Cellulase was adsorbed on the surface...

Preparation of Chitosan/Magnetic Porous Biochar as Support for Cellulase Immobilization by Using Glutaraldehyde

In this work, porous biochar was obtained from sugarcane bagasse by alkali activation and pyrolysis and then magnetized with γ-Fe2O3 by calcination. After functionalization with chitosan and activation with glutaraldehyde, the as-prepared chitosan/magnetic porous biochar served as a support to immobilize cellulase by covalent bonds. The immobilization amount of cellulase was 80.5 mg cellulase/g support at pH 5 and 25 °C for 12 h of immobilization. To determine the enzymatic properties, 1% carboxymethyl cellulose sodium (CMC) (dissolved in 0.1 M buffer) was considered as a substrate for hydrolysis at different pH values (3–7) and temperatures (30–70 °C) for 30 min. The results showed that the optimum pH and temperature of the free and immobilized cellulase did not change, which were pH 4 and 60 °C, respectively. The immobilized cellulase had a relatively high activity recovery of 73.0%. However, it also exhibited a higher Michaelis–Menten constant (Km) value and a slower maximum reaction velocity (Vmax) value compared to the free enzyme. In the reusability assay, the immobilized cellulase showed initial glucose productivity of 330.9 mg glucose/g CMC and remained at 86.0% after 10 uses. In conclusion, the chitosan/magnetic porous biochar has great potential applications as a support for enzyme immobilization.

Adsorption of Cellulase on Wrinkled Silica Nanoparticles with Enhanced Inter-Wrinkle Distance

Mesoporous silica materials offer a unique opportunity for enzyme immobilization thanks to their properties, such as tuneable pore size, large surface area and easy functionalization. However, a significant enhancement of cellulase enzyme activity entrapped inside the silica pores still represents a challenge. In this work, we immobilized cellulase by adsorption on wrinkled silica nanoparticles (WSNs), obtaining an active and stable biocatalyst. We used pentanol as co-solvent to synthesize WSNs with enhanced inter-wrinkle distance in order to improve cellulase hosting. The physical-chemical and morphological characterization of WSNs and cellulase/WSNs was performed by thermogravimetric (TG), Fourier transform infrared (FT-IR), and transmission electron microscopy (TEM) analyses. The obtained results showed that this matrix generates a favourable microenvironment for hosting cellulase. The results of the catalytic assays and operational stability confirmed the key role of size, morphology and distribution of the pores in the successful outcome of the cellulase immobilization process. The immobilization procedure used allowed preserving most of the secondary structure of the enzyme and, consequently, its catalytic activity. Moreover, the same value of glucose yield was observed for five consecutive runs, showing a high operational stability of the biocatalyst.

Biochemical Degradation of Chitosan over Immobilized Cellulase and Supported Fenton Catalysts

This paper describes the application of Fe-MCM-48 (Mobil Composition of Matter No.48) and cellulase-MCM-48 catalysts for the depolymerization of chitosan. The results show that H2O2 is a good oxidant for the depolymerization of chitosan in the presence of Fe-MCM-48. The average polymerization degree of the product decreased to 6.1, and decreased to 29.2 when cellulase-MCM-48 was used as a catalyst, because the effect of the enzyme was affected by the molecular structure of chitosan. When both materials were used for depolymerization, the average degree of polymerization sharply decreased to 3.8. The results show that the two degradation methods can promote each other to obtain oligosaccharides with a lower degree of polymerization. This provides a new method for the controllable degradation of chitosan and lays a good foundation for the industrial production of chitosan oligosaccharides with a low degree of polymerization.