Preparation method of water-soluble active polysaccharide in boletus edulis, and application thereof
A technology of delicacy boletus and active polysaccharides, which is applied to the preparation of water-soluble active polysaccharides in delicacy boletus thallus, the field of application of delicacy boletus polysaccharides in antibacterial, can solve the problem of long growth cycle, fruit body active substances Problems such as low yield and difficult cultivation
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Embodiment 1
[0037] A kind of preparation method of water-soluble active polysaccharide in boletus thalline, its steps are:
[0038] A Inoculate Boletus deliciosa (No. ACCC50559, provided by China Agricultural Microorganism Culture Collection Center) on a solid slant medium, and culture it at a constant temperature of 25°C for 6 days to obtain slant seeds. Solid slant medium (%): potato 20, glucose 2.0, yeast powder 1.4, agar 2, peptone 0.6, dihydrogen phosphate 0.1, zinc sulfate heptahydrate 0.05.
[0039] B Take 4-5 loops of the cultured slant and inoculate it into the primary liquid seed medium, add 10 glass beads in a 250ml Erlenmeyer flask with a liquid volume of 100ml, and incubate at 120r / min at 25°C for 60 hours. Each component and its content in the medium are (%): 20% potato, 2.0% glucose, 1.4% yeast powder, 0.6% peptone, 0.1% methyl dihydrogen phosphate, and 0.05% zinc sulfate heptahydrate.
[0040] C Inoculate the first-level liquid seeds after 60 hours of cultivation into the...
Embodiment 2
[0046] A kind of preparation method of water-soluble active polysaccharide in boletus thalline, its steps are:
[0047] Embodiment 2 steps A-E are the same as steps A-E in embodiment 1
[0048] F Take an appropriate amount of crushed boletus mycelium, add 50 times the amount of deionized water, and extract at 53 or 54 or 55 or 56°C for 2 hours. Repeat 3 times, combine the 3 extracts, and centrifuge the extract at 7000rpm for 8min. The filtrate was concentrated at 54°C under reduced pressure (0.08-0.09MPa) to 1 / 3 of the original volume, and 3 times the volume of 95% (v / v) ethanol was added to the concentrated extract with stirring to precipitate the polysaccharide in the concentrated extract , collect the crude polysaccharide precipitate of boletus mycelium; add deionized water to the crude polysaccharide precipitate to dissolve it, make it fully redissolved, use the sevag method to remove protein, and repeat until no protein is detected. After concentrating under reduced pre...
Embodiment 3
[0051] A kind of preparation method of water-soluble active polysaccharide in boletus thalline, its steps are:
[0052] Steps A-C of embodiment 3 are the same as steps A-C in embodiment 1
[0053] D Inoculate the secondary liquid seeds after 40 hours of cultivation into a 250mL Erlenmeyer flask for 72 hours of fermentation at an inoculum volume of 5%. The components of the fermentation medium are (%): yeast extract 1, peptone 2, and glucose 2.
[0054] E The mycelium is separated from the fermentation broth, rinsed with clean water, dried at 50°C, and ground to obtain mycelium dry powder.
[0055] F Take an appropriate amount of crushed boletus mycelium, add 50 times the amount of deionized water, and sonicate at 52 or 53 or 54 or 55 or 56°C for 8.5 minutes. Repeat 3 times, combine the 3 extracts, and centrifuge the extract at 7000rpm for 8min. The filtrate was concentrated at 54°C under reduced pressure (0.08-0.09MPa) to 1 / 3 of the original volume, and 3 times the volume o...
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