Large batch production of Galactooligosaccharides using β‐glucosidase immobilized on chitosan‐functionalized magnetic nanoparticle

Abstract

β-glucosidase (BglA) immobilization from Thermotoga maritima on magnetic nanoparticles (MNPs) functionalized with chitosan (Cs) were efficiently investigated to improve lactose conversion and galactooligosaccharides (GOS) production. We used a batch method in order to improve the conversion of lactose to GOS. The efficiency and yield of immobilization were 79% and immobilized BglA was effectively recycled via a magnetic separation procedure through a batch-wise GOS with no activity lessening. Furthermore, analyses were done through screening kinetics of enzyme activity, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Fourier transform infrared spectroscopy (FT-IR), and transmission electron microscopy (TEM). Proposed methodology of immobilization shows a potential application as it is stable which was proved through many methods including pH, temperature, heat treatment, storage, and kinetics of the enzyme. GOS and residual enzyme activity showed to be 28.76 and 40.44%, respectively. However, free enzyme synthesis of GOS yield was just 24% after 12 hr. This study proposed applying magnet in the immobilization process of BglA on Cs-MNPs to produce GOS as new method for immobilizing enzyme in a biostable and cost-efficient way. Practical applications This paper focus on immobilization of BglA from T. maritima onto MNPs functionalized with CS to investigate their further possibility improving lactose conversion and GOS production. Interestingly, a successful immobilization of Tm-BglA on the substrates were achieved in Cs-MNPs. The obtained results from enzyme activity, SDS-PAGE, FT-IR, and TEM showed that the high binding capacity of BglA to Cs-MNPs was successfully obtained. Furthermore, the binding efficiency calculation indicated that the immobilized BglA-Cs-MNPs conserved 40.44% of its native activity at the end of its 6th repeated use. In addition, magnetic separation technique was successfully employed for reuse of the immobilized BglA for repetitive batch-wise GOS without significant loss of activity. How to cite this article: Alnadari F, Xue Y, Almakas A, et al. Large batch production of Galactooligosaccharides using β-glucosidase immobilized on chitosan-functionalized magnetic nanoparticle.

Production and immobilization of β-glucanase from Aspergillus niger with its applications in bioethanol production and biocontrol of phytopathogenic fungi

“…The immobilization of enzyme led to an increase in enzyme rigidity, which is generally reflected by improving the stability against denaturation on rising the temperature 12 , 33 . The successful immobilization of β-glucanase on carrageenan and chitosan was reported by various investigators 8 , 16 , 34 .

Figure 5 Schematic representation of β-glucanase immobilization on activated carrageenan (A) and chitosan (B) beads.

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Immobilization and Recyclability of Β-Glucosidase From Thermatoga Maritima on Biopolymer-Coated Magnetic Nanoparticles

“…However, due to the fundamentally complicated nature of the enzyme structure, no single approach is optimal for all compounds or goals. The formulae may be used to determine the predicted immobilization yield (Alnadari et al 2021).…”

“…The recent interest in nanotechnology has provided a wide variety of nano scaffolds, which may support enzyme immobilization due to its potential applications in biotechnology and biomedicine (Ansari and Husain 2012). Immobilization of enzymes is useful for commercial applications owing to the simplicity of handling, ease of separating enzymes from reaction mixtures and reusing, a lower transfer resistance to solve the diffusion problem, lower operating cost, and a potential increase in heat and pH stability (Tischer and Wedekind 1999;Ansari and Husain 2012;Verma et al 2013;Vaghari et al 2016;Alnadari et al 2020;Alnadari et al 2021;Abdalmegeed et al 2022). Biological systems and their components have a significant impact because biotechnology involves nano-sized molecules such as proteins and nucleic acids.…”

Thiolation of Myco-Synthesized Fe3O4-NPs: A Novel Promising Tool for Penicillium expansium Laccase Immobilization to Decolorize Textile Dyes and as an Application for Anticancer Agent

“…In the present study, the immobilized laccase was more stable toward heat denaturation, with only a 5% of laccase activity loss at 50 • C when compared to the free counterpart (Figure 7C(b)). The enhanced thermostability of immobilized enzyme could be assigned to the decline of heat transfer to enzyme microenvironment and the protection of active conformational site as a result of the covalent linking arising between laccase and the thiol functionalized magnetic nanoparticles [6,41].…”

“…The Fe 3 O 4 /3-MPA-S-S-laccase exhibited remarkable stability while retaining at 84.34% of its initial activity after 10 cycles. The activity loss in repetitive cycles of substrate oxidation might be connected to the repetitive joining of the substrate to active sites of the biocatalyst and hence, influence the binding potency between the enzyme and carrier which is linked to denaturation and inactivation of the enzyme [6,41].…”