Immobilized enzyme catalyst, preparation method thereof and application of immobilized enzyme catalyst in synthesis of vitamin A palmitate
A technology for immobilizing enzymes and catalysts, which can be used in biochemical equipment and methods, enzymes, etc., which are immobilized on or in inorganic carriers, and can solve the problems of acid resistance, poor hydrophobicity and diffusivity, and achieve high physical and chemical stability. Use efficiency, enhance the effect of structural stability
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Embodiment 1
[0047] Preparation of immobilized enzyme catalyst:
[0048] Dissolve 5.0 g of free enzyme (Candida sp.99-125 dry enzyme powder) in 250 mL of 0.2 mol / L sodium hydrogen phosphate buffer, adjust the pH to 4.5, and store it under nitrogen atmosphere with stirring.
[0049] 10.0 g of tetrabutyl titanate was weighed and added to 100 mL of ethanol / acetic acid mixed solution (the volume ratio of ethanol to acetic acid was 0.5:1). Weigh 1g of α-cellulose and add it into 50mL of methanol, heat up to 30°C and stir vigorously to disperse evenly. Add the mixed solution of tetrabutyl titanate above to the cellulose methanol solution dropwise, and after adding dropwise for 1 hour, transfer the resulting mixture to an electrospinning syringe, and after electrospinning at 10kV for 5 hours, place the obtained solid in a nitrogen atmosphere , baked at 300°C for 4h.
[0050] Accurately weigh 10.0 g of the roasted solid and add it to the above-mentioned sodium hydrogen phosphate buffer solution ...
Embodiment 2
[0056] Preparation of immobilized enzyme catalyst:
[0057] Dissolve 5.0 g of free enzyme (Candida sp.99-125 dry enzyme powder) in 300 mL of 0.3 mol / L sodium hydrogen phosphate buffer, adjust the pH to 5.0, and store it under nitrogen atmosphere with stirring for later use.
[0058] Weigh 10.0 g of cobalt nitrate and add to 200 mL of ethanol / acetic acid mixed solution (the volume ratio of ethanol to acetic acid is 0.6:1). Weigh 2.0g of α-cellulose and add it into 400mL of methanol, heat up to 40°C and stir vigorously to disperse evenly. The above mixed solution of cobalt nitrate was added dropwise to the cellulose methanol solution, and after 1.2 h, the resulting mixture was transferred to an electrospinning syringe, and after electrospinning at 15 kV for 6 h, the resulting solid was placed in a nitrogen atmosphere, Calcined at 350°C for 4.5h.
[0059] Accurately weigh 10.0 g of the roasted solid and add it to the above-mentioned sodium hydrogen phosphate buffer solution con...
Embodiment 3
[0065] Modification and immobilized preparation of free enzyme:
[0066] Dissolve 5.0 g of free enzyme Candida sp.99-125 dry enzyme powder in 300 mL of 0.5 mol / L sodium hydrogen phosphate buffer, adjust the pH to 5.5, and store it under nitrogen atmosphere with stirring. 10.0 g of tetrabutyl titanate was weighed and added to 300 mL of ethanol / acetic acid mixed solution (the volume ratio of ethanol to acetic acid was 0.8:1). Weigh 3.0g of pectin and add it to 600mL of methanol, heat up to 60°C and stir vigorously to disperse evenly.
[0067] Add the mixed solution of tetrabutyl titanate above to the pectin methanol solution dropwise, and after 1.8h, transfer the resulting mixture to the syringe for electrospinning, and after electrospinning at 18kV for 8h, put the obtained solid under nitrogen In the atmosphere, bake at 450°C for 6.5h. Accurately weigh 10.0 g of the roasted solid and add it to the above-mentioned sodium hydrogen phosphate buffer solution containing free enzym...
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