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. 2024 Feb;47(2):249-261.
doi: 10.1007/s00449-023-02959-1. Epub 2024 Jan 10.

Covalent immobilization of β-galactosidase using a novel carrier alginate/tea waste: statistical optimization of beads modification and reusability

Mohamed A A Abdella  1 , Mohamed E Hassan  2   3
Affiliations

Covalent immobilization of β-galactosidase using a novel carrier alginate/tea waste: statistical optimization of beads modification and reusability

Mohamed A A Abdella et al. Bioprocess Biosyst Eng. 2024 Feb.

Abstract

β-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.

Keywords: Alginate/tea waste; Chemical modification; Immobilized β-galactosidase; Reusability; Statistical optimization.

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Conflict of interest statement

The authors declare that there is no conflict of interests.

Figures

Fig. 1
Fig. 1
Encapsulator for making uniform gel beads
Scheme 1
Scheme 1
Activation of alginate/tea waste complex and immobilization processes
Fig. 2
Fig. 2
A The relation between predicted and actual values B Normal probability plot of residuals C Plot of residuals vs. predicted values of β-galactosidase IY% using BBD for optimizing beads modifications
Fig. 3
Fig. 3
The 3D response surface and contour plots showing interactions between each two variables affecting beads modifications A PEI concentration and PEI pH B PEI concentration and GA concentration C PEI pH and GA concentration
Fig. 4
Fig. 4
FT-IR analysis of Alg/TW gel beads (Alg/TW), Alg/TW beads treated with PEI (Alg/TW/PEI), Aminated Alg/TW beads treated with GA (Alg/TW/PEI/GA), and Activated Alg/TW beads treated with β-gal enzyme (Alg/TW/PEI/GA/Enz)
Fig. 5
Fig. 5
TGA of Alg/TW gel beads (A), Alg/TW beads treated with PEI (B), Aminated Alg/TW beads treated with GA (C), and Activated Alg/TW beads treated with β-gal enzyme (D)
Fig. 6
Fig. 6
SEM of Alg/TW gel beads (A), Alg/TW beads treated with PEI (B), Aminated Alg/TW beads treated with GA (C), and Activated Alg/TW beads treated with β-gal enzyme (D)
Fig. 7
Fig. 7
Operational stability of immobilized β-galactosidase
See this image and copyright information in PMC

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