Effect of enzyme to support ratio on immobilization of β-glycosidase for synthesis of butyl glycoside / Nooranis Natasha Binti Abdullah

Faculty of Chemical and Process Engineering Technology

Final Year Report (Bachelor of Chemical Engineering ) -- Universiti Malaysia Pahang Al-Sultan Abdullah - 2025

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This study explores the effect of enzyme-to-support ratios β-glycosidase immobilization using modified chitosan-based supports. Chitosan beads were synthesized via emulsion cross-linking and modified with 80% butanal to enhance hydrophobicity (Chang et al., 2010). Characterization techniques such as thermogravimetric analysis (TGA), scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR) confirmed the structural integrity and hydrophobicity of the modified beads. The enzyme was immobilized on these supports, and its activity was assessed using p-nitrophenyl-β-D-glucopyranoside (pNPG) as a substrate. Hydrophobicity testing indicated that butanal modification improved enzyme binding and stability on the beads. Results showed that the enzyme-to-support ratio significantly impacted immobilization efficiency. The effect of the enzyme-to-support ratio was measured with five specific weight ratios are selected which are 1:0.5, 1:1, 1:2, 1:5, 1:7, and 1:10. These ratios represent the proportion of enzyme to micro chitosan in the mixture. For each ratio, the activated enzyme is mixed with activated micro chitosan and kept under mild stirring at 150 rpm to ensure uniform interaction between the enzyme and the support. The process is conducted at 25°C for 3 hours, which allows adequate time for the adsorption process to occur. Immobilization of β-glycosidase at ratio (1:0.5) show the highest yield of 17.5 % successfully bound when been introduced with 30 U of β-glycosidase into the system proving longer alkyl chain increase the ability of the support to immobilize enzyme(Almulaiky et al., 2023).