Method for preparing chitin oligosaccharide or chitosan oligosaccharide by using immobilized enzyme technology
A technology of immobilizing enzymes and chitooligosaccharides, applied in the field of chemical biology of renewable resources, can solve the problems of non-reusable, harsh reaction conditions, poor stability, etc., and achieve high affinity, good mechanical properties, and stable chemical properties.
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Embodiment 1
[0013] Embodiment 1: first the FeSO of 0.5mol / L 4 ·7H 2 O and 0.5mol / L FeCl 3 ·6H 2 O was mixed at a molar ratio of 1:2, 5 mol / L NaOH was added, and vigorously stirred until an oily magnetic fluid suspension was obtained. Dissolving chitosan in an acetic acid solution with a volume percentage concentration of 1-5% is made into a chitosan solution with a weight-to-volume ratio of 3%. In 50mL of the chitosan solution, add 3mL of the magnetic fluid and stir evenly. Add 50mL of liquid paraffin, drop in the emulsifier span80, stir at high speed to make the system into an emulsion, add glutaraldehyde with a volume of 1% of the system volume as a crosslinking agent, and pour the emulsion that has fully reacted at 35-45°C into the other In 5 times the volume of NaOH / ethanol (1:1) mixture, keep stirring for a period of time, discard the oil layer and water layer, wash off the adsorbate on the particle surface, and obtain a black magnetic shell with a particle size of 20-30um and a r...
Embodiment 2
[0014] Embodiment 2: first the FeSO of 0.25mol / L 4 ·7H 2 O and 0.25mol / L FeCl 3 ·6H 2 O was mixed at a molar ratio of 1:2, 5 mol / L NaOH was added, and vigorously stirred until an oily magnetic fluid suspension was obtained. Dissolve chitosan in an acetic acid solution with a concentration of 1-5% by volume to form a chitosan solution with a weight-volume ratio of 4%. Add 10mL of magnetic fluid to 50mL of the chitosan solution, stir evenly, and add 50mL of liquid paraffin, drop emulsifier span80, stir at high speed to make the system into an emulsion, add a volume ratio of 3% glutaraldehyde as a cross-linking agent, and pour the fully reacted emulsion at 35-45°C into 5 times the volume of NaOH / ethanol (1:1) mixed solution, keep stirring for a period of time, discard the oil layer and water layer, wash off the adsorbate on the surface of the particles, and obtain black magnetic chitosan microspheres with a more uniform particle size. Dissolve the cellulase with a weight rat...
Embodiment 3
[0015] Embodiment 3: first the FeSO of 1mol / L 4 ·7H 2 O and 1mol / L FeCl 3 ·6H 2 O was mixed at a molar ratio of 1:2, 5 mol / L NaOH was added, and vigorously stirred for 30 min to obtain an oily magnetic fluid suspension. 50mL 4% (w / v) chitosan solution, mixed with 3mL magnetic fluid, stirred evenly, added 50mL liquid paraffin, dripped emulsifier span80, stirred at high speed to make the system emulsion-like, added 4% glutaraldehyde by volume as Cross-linking agent, pour the fully reacted emulsion at 35-45°C into 10 times the volume of NaOH / ethanol (1:1) mixture, keep stirring for a period of time, discard the oil layer and water layer, and wash away the particles on the surface Adsorbate, to obtain black magnetic chitosan microspheres with uniform particle size. Dissolve cellulase with a weight ratio of 0.3 to the immobilized carrier in 0.05-0.4mol / L, HAc-NaAc buffer solution with a pH of 4.6-5.6, shake fully at room temperature, filter to remove the enzyme solution, and th...
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