Enterococcus monteilii and application thereof in fermentation of Chinese herbal medicines
A technology for Enterococcus montelii and fermentation of Chinese herbal medicines, applied in the field of microorganisms, can solve the problems of low output and can not be fully utilized, etc., and achieves the effects of simple operation, broad practical application prospects and strong practicability.
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Embodiment 1
[0027] Isolation and culture of strains derived from cow dung:
[0028] (1) Sample collection and processing
[0029] Prepare 12 sterile 10mL glass sample collection tubes, collect Simmental (9 heads), Charolais (2 heads), and Black and White Flowers (1 head), a total of 12 cow feces samples of three breeds. The feed these cattle eat includes whole price concentrate, whole plant silage, hay, corn, wheat bran and Mushengyuan (fermented traditional Chinese medicine). Collect and accurately weigh 0.5 g of feces samples from each of the above 12 cows, add 9.5 mL of sample diluent (0.9 % mass concentration of normal saline), and add numbers 1-1, 1-2, 1-3, 2-1, 2-2, 2-3, 3-1, 3-2, 3-3, 4-1, 4-2, 4-3 in the sample collection tube and shake quickly to mix. were diluted to 10 -5 、10 -6 、10 -7 multiples of three.
[0030] MRS agar medium
[0031] Peptone 10g / L, beef extract powder 5g / L, yeast extract powder 4g / L, glucose 20g / L, Tween-1ml / L, dipotassium hydrogen phosphate 2g / L, a...
Embodiment 2
[0040] Screening strains that can increase the content of active components in Astragalus membranaceus
[0041] Astragalus decoction pieces are Mongolian Astragalus. Astragalus was naturally dried, crushed to 60 mesh, and stored for later use.
[0042] (1) Recovery and passage of strains
[0043] Prepare MRS medium, and use high pressure for standby. Inoculate 1mL of bacterial liquid into 50mL of prepared MRS liquid medium according to the proportion of 2% of the 12 isolated bacterial strains stored in the freezer, and incubate anaerobically at 37°C for 24h. The resuscitated strains continued to be inoculated into new MRS liquid medium at a ratio of 2%, and thus passed to the fourth generation.
[0044] (2) Preparation and fermentation of fermentation medium
[0045] Prepare 12 groups of fermentation media of 2 L each, and set up 3 replicates. Take 200 g of crude drug Astragalus root as the control group, and compare the content changes of the main components of Astragalus...
Embodiment 3
[0061] The preparation embodiment of the fermented bacterial agent that comprises bacterial strain MY-YD-1
[0062] Strain MY-YD-1 (inoculum size is 10 3 CFU) into the slant of MRS medium for cultivation: time: 16-24 hours, environment: anaerobic fermentation, temperature: 36.5-37.5°C, initial pH: 6.0-6.4.
[0063] The composition of MRS slant medium is: 1% peptone, 1% beef extract, 0.5% yeast powder, 0.2% diamine citrate, 2% glucose, 0.5% sodium acetate, 0.2% dipotassium hydrogen phosphate, 0.0058% MgSO 4 ·7H 2 O, 0.025% MnSO 4 4H 2 O, 0.1% (v / v) Tween-80, pH 6.2, 3% agar.
[0064] The culture obtained in the previous step (the inoculum size was 10 3 CFU) is inserted into primary or secondary seed medium for cultivation: time 24-48 hours, environment: anaerobic fermentation, temperature: 36.5-37.5°C, rotation speed 100r / min, initial pH: 6.0-6.4, and fermentation is obtained after cultivation Bacteria.
[0065] The composition of the seed medium is: 1% peptone, 1% beef...
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