A kind of horse milk with hypolipidemic acid prepared by mixed fermentation of compound strains and preparation method thereof
A technology of mixed fermentation and compound strains, applied in the field of microbial fermentation, to achieve the effect of wide practicability and development value
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Embodiment 1
[0055] Embodiment one: Kluyveromyces marxe ( Kluyveromyces marxianus ) Screening, separation, purification and identification of WWMJ-1CGMCCNo.8432
[0056] Primary screening: 1 mL of traditional fermented horse milk is put into a test tube equipped with 9 mL of sterile saline, and fully mixed evenly on a vortex mixer. Dilute the fermented sour horse milk in 10-fold steps, and the dilution is 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 . Use a pipette to draw 1mL of each of the above dilutions, pour into the YPD medium by the pouring method, and place the plate in an incubator at 28°C for 24-48h. Observe and record the characteristics of the colonies and pick out suspected yeast colonies, and number the suspected strains.
[0057] Purification of strains: Inoculate the suspected strains on YPD agar medium again, culture them at a constant temperature in a 28°C incubator for 24-48h, and repeat 3-4 times to purify the yeast. The purified yeast strains were inoculated on ...
Embodiment 2
[0063] Embodiment two: the selection of culture medium
[0064] MRS medium: glucose 20g, peptone 10g, beef extract 10g, yeast extract 5g, diamine citrate 2g, dipotassium hydrogen phosphate 2g, sodium acetate 5g, magnesium sulfate 0.58g, manganese sulfate 0.25g, Tween-801mL, distilled water 1000mL.
[0065] YPD medium: yeast extract 10g, peptone 20g, glucose 20g, distilled water 1000mL.
[0066] Wort medium: malt extract powder 130g, chloramphenicol 0.1g, distilled water 1000mL.
[0067] The solid medium of each of the above-mentioned mediums adopts the corresponding addition of 20.0 g / L of agar.
[0068] Kluyveromyces marx ( Kluyveromyces marxianus) Physiological and biochemical test medium of WWMJ-1CGMCCNo.8432: Nitrate reduction medium: 3g of bovine extract, 5g of peptone, 1g of potassium nitrate, 1000mL of distilled water, dissolve each component in distilled water, adjust the pH to 7.2-7.4, and pack into test tubes , about 5mL per tube, sterilized at 121°C for 15min...
Embodiment 3
[0069] Example 3: Mixed fermentation of compound strains to prepare sour horse milk
[0070] See attached figure 1 , using self-selected Kluyveromyces marx ( Kluyveromyces marxianus ) WWMJ-1 and selected Lactobacillus casei ( Lactobacillus casei ), Lactococcus lactis ( Lactococcus lactis ) and Monascus ( Monascus ruber ) to prepare the method for sour horse milk by fermenting the composite strain of the composition, the specific preparation method steps are as follows.
[0071] (1) Horse milk pretreatment: filter fresh horse milk to remove impurities and foreign matter in horse milk, then preheat horse milk to 60-65°C, homogenize at 20MPa for 10-15min, and after homogenization, horse milk Milk is pasteurized and used for fermentation; the amount of mother’s fermented liquid, fermented liquid 1, and fermented liquid 2 that needs to be pretreated is 3:20:20 by volume.
[0072] (2) Preparation of seed fermentation liquid and mother fermentation liquid.
[0073] Seed f...
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