Mesoporous titanium dioxide immobilized enzyme and preparation method and application thereof
A technology of mesoporous titanium dioxide and immobilized enzymes, which is applied in the fields of immobilization on or in inorganic carriers, chemical industry, sustainable manufacturing/processing, etc. It can solve the problems of unreported immobilization methods and achieve good reusability , Applicable pH and temperature range expansion, the effect of structural stability
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Embodiment 1
[0032] M-TiO 2 Balance of carrier: weigh 5g M-TiO 2 For the mesoporous material, the carrier was equilibrated with 250 mL of Tris-HCl buffer solution of pH 8.050 mmol / L for 24 hours, and stored in the above buffer solution at 4° C. for future use.
[0033] Mesoporous material M-TiO 2 Immobilization of γ-glutamyl transpeptidase on the carrier: the above M-TiO 2 Drain, weigh 1gM-TiO 2 Put the wet granules in a sand core tube, add 10 mL of 0.038 mg / mL γ-glutamyl transpeptidase solution (enzyme activity is 22.35 U / mL), and shake and react in an ice bath at 4°C for 2.5 hours. After the reaction, wash off the excess enzyme solution with a Tris-HCl buffer solution of pH 8.050mmol / L, and store it in the above buffer solution to obtain M-TiO 2Immobilized γ-glutamyl transpeptidase. Its enzyme activity is 131U / g carrier, and the recovery rate of enzyme activity is 58.6%.
Embodiment 2
[0035] M-TiO 2 Balance of carrier: weigh 5g M-TiO 2 For the mesoporous material, the carrier was equilibrated with 250 mL of Tris-HCl buffer solution of pH 8.050 mmol / L for 24 hours, and stored in the above buffer solution at 4° C. for future use.
[0036] Mesoporous material M-TiO 2 Immobilization of γ-glutamyl transpeptidase on the carrier: the above M-TiO 2 Drain, weigh 1gM-TiO 2 The wet granules were placed in a sand core tube, 10 mL of 0.073 mg / mL γ-glutamyl transpeptidase solution (enzyme activity: 22.62 U / mL) was added, and the reaction was shaken in an ice bath at 4°C for 4 hours. After the reaction, wash off the excess enzyme solution with a Tris-HCl buffer solution of pH 8.050mmol / L, and store it in the above buffer solution to obtain M-TiO 2 Immobilized γ-glutamyl transpeptidase. Its enzyme activity is 119U / g carrier, and the recovery rate of enzyme activity is 52.6%.
Embodiment 3
[0038] M-TiO 2 Balance of carrier: weigh 5g M-TiO 2 For the mesoporous material, the carrier was equilibrated with 250 mL of Tris-HCl buffer solution of pH 8.050 mmol / L for 24 hours, and stored in the above buffer solution at 4° C. for future use.
[0039] Mesoporous material M-TiO 2 Immobilization of γ-glutamyl transpeptidase on the carrier: the above M-TiO 2 Drain, weigh 1gM-TiO 2 Put the wet granules in a sand core tube, add 10 mL of 0.017 mg / mL γ-glutamyl transpeptidase solution (enzyme activity is 21.07 U / mL), and shake and react in an ice bath at 4°C for 2.5 hours. After the reaction, wash off the excess enzyme solution with a Tris-HCl buffer solution of pH 8.050mmol / L, and store it in the above buffer solution to obtain M-TiO 2 Immobilized γ-glutamyl transpeptidase. Its enzyme activity is 107.8U / g carrier, and the recovery rate of enzyme activity is 51.15%.
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