Effect of surface charge conditions of carriers on the immobilization of β-d-glucosidase

In this study, a series of acidic or alkaline polypeptide chains were designed and grafted onto DEG-AM resin using Fmoc solid-phase synthesis to study the relationship between enzyme conformation and carrier surface charge. β-d-glucosidase (βGase) was then immobilized onto these modified carriers by adsorption. Each form of immobilized βGase showed decreasing specific activity compared to that of the free. It could be attributed to both the changes in the enzyme conformation and the decrease in mass transfer efficiency. The optimum temperature of free βGase, DEG@B3-βGase is 55 °C, which of DEG@A3-βGase is 65 °C and they all have the highest activity at pH 5. The E_a values ​​of free βGase, DEG@A3-βGase, and DEG@B3-βGase are 0.546 kJ/mol, 0.224 kJ/mol, and 0.446 kJ/mol, and the K_m values were 1.30 mmol/L, 1.44 mmol/L and 2.63 mmol/L, respectively. It shows that free βGase and DEG@A3-βGase are more similar. Meanwhile, the free βGase (1.0 g/L, pH 5.0) stored at 4 °C has a shorter half-life (t_1/2), which is only 9 days. However, the half-life of DEG@B3-βGase and DEG@A3-βGase is 20 days and over 60 days, indicating that the negative charged surface was conducive to maintenance of the structure and catalytic property of βGase.