“…Enzyme structural alterations or adsorption of the substrates and products, which affect the reaction sites, may have decreased the activity during repeated cycling [ 41 , 42 ]. This is consistent with previous studies showing that sodium alginate-coated magnetic nanoparticles [ 43 ] and modified polyester [ 16 ] used as β-glucosidase carrier supports maintained 39.5 % and 82 % of the original activities after 10 and 6 cycles, respectievly. Fig.…”
“…Enzyme structural alterations or adsorption of the substrates and products, which affect the reaction sites, may have decreased the activity during repeated cycling [ 41 , 42 ]. This is consistent with previous studies showing that sodium alginate-coated magnetic nanoparticles [ 43 ] and modified polyester [ 16 ] used as β-glucosidase carrier supports maintained 39.5 % and 82 % of the original activities after 10 and 6 cycles, respectievly. Fig.…”
“…The effect of pH on the activity of immobilized β-glucosidase in the single-channel microreactor was studied and compared to that of the free enzyme (Figure 4). Both enzyme forms exhibit maximum activity at a pH value of 7.0 like that previously reported in the literature [38][39][40][41][42][43]. When the pH value rises to 8.0 and 9.0, the immobilized enzyme excels in activity preservation over the free form, indicating a stabilizing effect of the microreactor towards the protein molecules [41,42].…”
Section: Biocatalytic Characterization Of the Immobilized Single-chan...supporting
Microfluidic devices have attracted much attention in the current day owing to the unique advantages they provide. However, their application for industrial use is limited due to manufacturing limitations and high cost. Moreover, the scaling-up process of the microreactor has proven to be difficult. Three-dimensional (3D) printing technology is a promising solution for the above obstacles due to its ability to fabricate complex structures quickly and at a relatively low cost. Hence, combining the advantages of the microscale with 3D printing technology could enhance the applicability of microfluidic devices in the industrial sector. In the present work, a 3D-printed single-channel immobilized enzyme microreactor with a volume capacity of 30 μL was designed and created in one step via the fused deposition modeling (FDM) printing technique, using polylactic acid (PLA) as the printing material. The microreactor underwent surface modification with chitosan, and β-glucosidase from Thermotoga maritima was covalently immobilized. The immobilized biocatalyst retained almost 100% of its initial activity after incubation at different temperatures, while it could be effectively reused for up to 10 successful reaction cycles. Moreover, a multi-channel parallel microreactor incorporating 36 channels was developed, resulting in a significant increase in enzymatic productivity.
“…One of the most important advantages of immobilization process is reusability that should be considered for industrial applications due to the possibility of reusing the enzyme over a long time. The immobilization strategy of an enzyme to a support material helps in recycling it for several batches and subsequently reducing the cost of reaction system [ 37 , 38 ]. The novel activated carrier Alg/TW was utilized to obtain a stable immobilized β-gal that can be separated easily from its products and restarted several reaction cycles.…”
Abstractβ-galactosidase has been immobilized onto novel alginate/tea waste gel beads (Alg/TW) via covalent binding. Alg/TW beads were subjected to chemical modification through amination with polyethyleneimine (PEI) followed by activation with glutaraldehyde (GA). Chemical modification parameters including PEI concentration, PEI pH, and GA concentration were statistically optimized using Response Surface methodology (RSM) based on Box–Behnken Design (BBD). Analysis of variance (ANOVA) results confirmed the great significance of the model that had F value of 37.26 and P value < 0.05. Furthermore, the R2 value (0.9882), Adjusted R2 value (0.9617), and predicted R2 value (0.8130) referred to the high correlation between predicted and experimental values, demonstrating the fitness of the model. In addition, the coefficient of variation (CV) value was 2.90 that pointed to the accuracy of the experiments. The highest immobilization yield (IY) of β-galactosidase (75.1%) was given under optimized conditions of PEI concentration (4%), PEI pH (9.5), and GA concentration (2.5%). Alg/TW beads were characterized by FT-IR, TGA, and SEM techniques at each step of immobilization process. Moreover, the immobilized β-galactosidase revealed a very good reusability as it could be reused for 15 and 20 consecutive cycles keeping 99.7 and 72.1% of its initial activity, respectively. In conclusion, the environmental waste (tea waste) can be used in modern technological industries such as the food and pharmaceutical industry.
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