Pectinase immobilizing carrier, preparation thereof, and method for immobilizing pectinase
A technology for immobilizing a carrier and pectinase, which is applied in the field of enzyme engineering, can solve problems such as no carrier, and achieve the effects of preventing environmental pollution, high enzyme activity preservation rate, and improving use efficiency.
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Embodiment 1
[0050] Stir and disperse with deionized water and soluble starch to form starch milk with a concentration of 30%. Weigh FeCl at a ratio of 1:1.5 3 , FeCl 2 , dissolved in deionized water to 10 times the starch milk, poured into the starch milk, and placed in a 65°C water bath to raise the temperature. After reaching the temperature, 0.5M NaOH solution was added dropwise to adjust the pH value to 10, and ultrasonic wave treatment was performed at 25KHz and 900W power for 30min. Place the treated liquid in a 65°C water bath and stir for 1.5 hours. After the reaction, let it stand and cool to room temperature, neutralize it with glacial acetic acid, wash it three times with 95% ethanol, and use a magnet with a residual magnetization of 12.3T The magnetic starch microspheres are separated from the liquid, the supernatant is poured off, and vacuum-dried at -45° C. and 20 Pa to obtain the magnetic starch microsphere carrier. The carrier is detected by laser diffraction particle s...
Embodiment 2
[0052] Stir and disperse with deionized water and cornstarch to form starch milk with a concentration of 40%. Weigh Fe at a ratio of 1:2 2 (SO 4 ) 3 , FeSO 4 , dissolved in deionized water to 15 times the starch milk, poured into the starch milk, and placed in a 65°C water bath to raise the temperature. After reaching the temperature, 0.5M KOH solution was added dropwise to adjust the pH value to 11, and ultrasonic treatment was performed at 20KHz and 450W power for 20min. Place the treated liquid in a 65°C water bath and stir for 2 hours. After the reaction, let it stand and cool to room temperature, neutralize it with glacial acetic acid to neutrality, wash it with 95% ethanol three times, and use a magnet with a residual magnetization of 12.3T. The magnetic starch microspheres are separated from the liquid, the supernatant is poured off, and vacuum-dried at -45°C and 20Pa to obtain the magnetic starch microsphere carrier. Detected by a laser diffraction particle size a...
Embodiment 3
[0054] Weigh 1 part of activated magnetic carrier cross-linked with 5% glutaraldehyde, 3 parts of 1% Penicillium oxalicum pectinase solution from goose source, adjust the volume to 25 times the volume with buffer solution with pH 4, and store at 30°C and Shake and react in a shaker at 200r / min for 3 hours, separate the magnetic starch microspheres from the liquid with a magnet with a residual magnetization of 12.3T, pour off the supernatant, and wash repeatedly with buffer until no protein is detected in the supernatant Until then, vacuum freeze-drying at -20°C to -45°C and 20 to 100Pa to obtain magnetically immobilized pectinase. Using laser diffraction particle size analyzer, scanning electron microscope, Fourier transform infrared spectrometer, and X-ray diffractometer, it can be seen that the particle size of immobilized pectinase is larger than that of magnetic starch microsphere carrier, and the functional group has changed, indicating that pectinase Has been cross-linke...
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