Preparation, purification and content detection methods for cerebroside
A technology for separation and purification of cerebrosides, applied in chemical instruments and methods, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of high cost, low output, small scale, etc., and achieve low cost and reliable quality The effect of control and short cycle
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Embodiment 1
[0068] Bacteria used: Tuber melanosporum
[0069] 1. Preparation of truffle mycelium containing cerebroside in the first stage:
[0070] 1), slant strain cultivation: the formula of the slant strain culture medium adopted in the present embodiment is: glucose 20 grams / liter, magnesium sulfate 2 grams / liter, potassium dihydrogen phosphate 2 grams / liter, agar 20 grams / liter 1. 1.0 liter of potato extract; the sterilization conditions are: 121° C. for 20 minutes. The culture temperature is 30°C, and the culture time is 7 days;
[0071] 2), first-level liquid seed culture: the medium formula is glucose 300 g / L, peptone 60 g / L, yeast extract 60 g / L, magnesium sulfate 20 g / L, potassium dihydrogen phosphate 30 g / L; pH value 8.0; (2) culture conditions: culture temperature 35 ℃, shaker speed 250 rev / min;
[0072] The first-level liquid seed culture can be carried out by transferring the slant bacteria into the shake flask equipped with the first-level liquid seed medium. The cultu...
Embodiment 2
[0086] Chinese truffles (Tuber sinense), Indian truffles (Tuber indium), black spore truffles (Tuber melanosporum), white truffles (Tuber magnatum ) and summer truffles (Tuber aestivum ) were subjected to liquid fermentation at different scales. The composition and culture conditions of the fermentation medium are shown in Table 1. Slant strain cultivation, primary liquid seed cultivation and secondary liquid seed cultivation are the same as in Example 1. The detection method of cerebroside content is the same as in Example 1. The yields of cerebrosides in the obtained truffle fermentation liquid and mycelia are shown in Table 2.
[0087] Table 1. Medium composition and culture conditions of different fermentation scales
[0088]
[0089] Table 2. Comparison of cerebroside production under different fermentation scale conditions
[0090]
Embodiment 3
[0092] Change the concentration and culture temperature of the main components of the slant culture medium of Tuber melanosporum (Tuber melanosporum) (see Table 3), and investigate its influence on the production of cerebroside by liquid fermentation of Tuber melanosporum. Primary liquid seed cultivation, secondary liquid seed cultivation and liquid fermentation are the same as in Example 1. The detection method of cerebroside content is the same as in Example 1. The yields of cerebrosides in the obtained tuber melanosporum fermentation broth and mycelia are shown in Table 3.
[0093] Table 3. The effect of slant strain culture on the production of cerebroside by liquid fermentation of Niger truffle
[0094]
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