Construction of Multienzyme Co-immobilized Hybrid Nanoflowers for an Efficient Conversion of Cellulose into Glucose in a Cascade Reaction

Abstract

Today, we are seeking an efficient biotransformation of cellulosic material into sustainable biochemical products to meet the increasing global energy demand. Herein, we report the fabrication of multienzyme hybrid nanoflowers (ECG-NFs) by co-immobilizing three recombinant enzymes (cellobiohydrolase (CBH), endo-glucanase (EG), and β-glucosidase (BG)) integrating a binary tag composed of elastin-like polypeptide (ELP) and His-tag to act as a tri-enzyme biocatalyst, which catalyzes the hydrolysis of cellulose into glucose. The prepared ECG-NFs exhibited excellent performance in terms of pH stability, thermal stability, storage stability, and catalytic efficiency compared to free multienzyme system. Notably, ECG-NFs could be recycled for up to eight consecutive runs. The K m and k cat/K m values for ECG-NFs were 9.33 g L–1 and 0.0051 L min–1 g–1, respectively, which were better than those of the free multienzyme system, indicating a better substrate affinity. Finally, the overall enzyme activity of ECG-NFs increased by 1.12 times and the degradation efficiency of ECG-NFs was superior to the free multienzyme system, which revealed that ECG-NFs could facilitate an effective one-pot hydrolysis of cellulose into glucose.

Cellulase Immobilization on Nanostructured Supports for Biomass Waste Processing

“…An original method of preparation of well-defined multienzyme hybrid nanoflowers (ECG-NFs) was proposed by Han et al by cross-linking all three cellulase enzymes (cellobiohydrolase (CBH), endo-glucanase (EG), and β -glucosidase (BG)) and combining a binary tag consisting of elastin-like polypeptide (ELP) and His -tag [ 79 ]. Here, recombinant enzymes (EG-Linker-ELP-His (EGLEH), CBH-Linker-ELP-His (CBHLEH), and Glu-Linker-ELP-His (GLEH) ( Figure 4 ), were assembled by incorporating a dual sticker (ELP-His) into the above enzymes.…”

“… Schematic of the formation mechanism of ECG-NFs and the functional mechanism of cellulase. Reproduced with permission from [ 79 ], American Chemical Society, 2021. …”

Facile Multilayer Deposition of Polyethylenimine and κ-Carrageenan on a 3D-Printed Lattice for Cellulase Immobilization

“…47 Moreover, certain free and immobilized cellulases at pH 3−4 with activity retentions of ∼35−40% were also reported in some previous works, which might be due to the nature of the enzyme under extremely acidic conditions. 48,49 Nevertheless, the E-GA-PkG showed satisfactory activity retention across the range of pH 5−7, with activity retention of 81.82 ± 5.43% at pH 7.…”

“Microflower-Templated” Janus Sheets: Synthesis and Application in Stabilizing Foams

“…So far, various immobilized enzymes have been widely prepared by biomimetic mineralization to form “microflowers” and applied to biodegradation. According to our previous work, we had prepared various enzyme-inorganic hybrid nanoflowers as biological catalyst; the enzymes mainly included β-glucosidase, , horseradish peroxidase, , glucose oxidase, glucoamylase, cellulase, bromelain, and glucose isomerase, which indicated that “microflowers” have the advantages of simple synthesis, mild reaction, regular morphology, high stability, and high specific surface area. Therefore, “microflowers” have become an ideal biomaterial template.…”