Water-soluble Chitooligosaccharide preparation method
A chitooligosaccharide and water-soluble technology is applied in the field of preparation of water-soluble chito-oligosaccharide, which can solve problems such as complicated processes, and achieve the effects of simple process and good tumor-inhibiting activity.
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Embodiment 1
[0010] Example 1: Take 100 grams of crab shell chitosan with a degree of deacetylation of 80.2%, dissolve it with 2 liters of 2% acetic acid solution, adjust the pH of the solution to 5.5 with saturated sodium hydroxide, control the temperature at 47°C, and add 10 milliliters of hemicellulose Suzyme crude enzyme solution catalyzes the degradation of chitosan. When the reaction solution is dripped into 2% sodium hydroxide solution without turbidity, use NMWL 10kDa ultrafiltration membrane to remove enzymes, and the solution that passes through the 10kDa ultrafiltration membrane is further fractionated with 3kDa and 1kDa ultrafiltration membranes to obtain solutions of three components . CHU10-3 is the fraction that passed through 10kDa membrane but failed to pass through 3kDa membrane; CHU3-1 is the fraction that passed through 3kDa membrane but failed to pass through 1kDa membrane. After the filtrate was concentrated in vacuo, the pH solution was adjusted to 8-9 with NaOH, se...
Embodiment 2
[0016] Example 2: Add 50 grams of shrimp shell chitosan with a deacetylation degree of 71.2% into 2 liters of 2% acetic acid solution and stir for 8 hours, then filter, adjust the pH of the solution to 5.0, control the temperature at 40°C, and add lysozyme to catalyze the degradation of chitosan sugar. After 24 hours, microfiltration was performed first, and then the oligosaccharide fraction was separated by ultrafiltration with a NMWL 1kDa ultrafiltration membrane. The component solution that did not pass was adjusted to pH 8-9 with saturated sodium hydroxide, centrifuged, and the supernatant was concentrated in vacuum, then ethanol sedimented, washed with ethanol, and dried to obtain a water-soluble chitooligosaccharide product with a yield of 38.7%. The average degree of polymerization is 21, and the degree of dispersion is 2.65.
Embodiment 3
[0017] Example 3: Take 25 grams of shrimp chitosan with a degree of deacetylation of 73%, add it to 1 liter of 2% lactic acid solution and stir for 8 hours, then filter it, adjust the pH of the solution to 5.5, control the temperature at 45°C, and add chitosan-containing enzyme The complex enzyme with hemicellulase catalyzes the degradation of chitosan. When the reaction solution is dripped into 2% sodium hydroxide solution without turbidity, use NMWL 10kDa ultrafiltration membrane to remove the enzyme, and the solution passing through the 10kDa ultrafiltration membrane is further NMWL The oligosaccharide fraction was separated by ultrafiltration with a 1kDa ultrafiltration membrane. Passed through 10kDa membrane, the component that does not pass through 1kDa membrane, obtains chitooligosaccharide lactic acid salt sample after freeze-drying, yield 23%, average polymerization degree 25, dispersion degree 2.85; After the enzyme, the chitooligosaccharide lactate sample was obtain...
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