A method for extracting and separating and purifying mannose oligosaccharides from yeast
A sugar oligosaccharide and yeast technology, applied in the field of extraction and separation and purification of mannose oligosaccharides, can solve the problems of separation and purification difficulties, limited promotion and application, and purification difficulties, and achieve the effect of improving production efficiency and reducing costs
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Embodiment 1
[0033] Accurately weigh 300 g of purified yeast and add it to a 2000 mL three-necked flask, put the flask in a vacuum drying oven at 50°C for 12 h, take out the flask from the vacuum drying oven, add 800 mL of acetic anhydride to it, and fix the flask in the oil Make sure that the liquid level of the reactant is immersed below the liquid level of the silicone oil in the oil bath, set the temperature of the oil bath to 90°C, add mechanical stirring, and set the stirring rate to 750 r / min. Slowly add 80 mL of concentrated sulfuric acid dropwise to the flask, and start timing when the temperature of the oil bath reaches and stabilizes at 90°C after the addition of concentrated sulfuric acid is completed, and stop heating when the reaction reaches 10 h. The flask was removed from the oil bath as a whole, and after cooling, the mixture in the flask was quickly poured into 5 L of deionized water and kept stirring vigorously, and allowed to stand still to allow the product to precipit...
Embodiment 2
[0039] Embodiment 2 adopts the same experimental procedure as in Example 1, and the difference is that the volume of the added vitriol oil is 60mL, the reaction time is 8 h, and the R of the product to be collected during the rough separation of the chromatographic column f The value is 0.3~0.4, and the product obtained after further purification is fully acetyl-protected mannotetraose, and 1~2 g of mannotetraose (Manα1-3Manα1-2Manα1-2Man) can be obtained after methanol / sodium methoxide deprotection.
[0040] Depend on Figure 1-4 It can be seen that through the separation and purification of the above steps, the purification of mannotriose and mannotetraose is basically achieved. The anomeric carbon of the polysaccharide is clearly visible in the figure 1 H and 13 C signal, due to the polysaccharide backbone in the hydrogen spectrum 1 The complex overlapping of the H signals is serious and cannot be assigned one by one. The signal assignments of the carbon spectrum are cle...
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