Microecological preparation-tetragenous viable bacteria preparation and method for preparing same
A live bacteria preparation and live bacteria technology, applied in the field of micro-ecological preparations, can solve the problems of unsustainable effect and single effect
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Embodiment 1
[0116] Example 1: Pure culture of live bacteria
[0117] (1) Pure cultivation of Bacillus subtilis (Bacillus subtilis CGMCC1.1414)
[0118] Strain activation
[0119] Open the freeze-dried strain of Bacillus subtilis aseptically, inoculate it on the nutrient broth agar slant of a test tube with a streak, culture at 37°C for 18-24h, then streak it and transfer it to the nutrient broth agar eggplant bottle slant, cultivate at 37°C for 20-24h , Microscopic examination, when more than 90% of the bacteria form spores, it is mature and ready to transplant.
[0120] Seed culture
[0121] Prepare a sterile conical flask containing glass beads, scrape off the mature bacterial sludge on the slope of the eggplant bottle with sterile water, put it into the conical flask, shake and disperse the bacterial sludge to obtain a uniform bacterial suspension. Heat the bacterial suspension in a water bath at 85°C for 5 minutes, and insert a 5% inoculum volume to 0.1m 3 Seed pot. The filling factor of ...
Embodiment 2
[0156] Example 2: Mixed culture of Clostridium butyricum, Lactobacillus casei and live yeast cells
[0157] Strain activation
[0158] The activation of the lyophilized species of Clostridium butyricum, Lactobacillus casei, and yeast is the same as described in Example 1.
[0159] Seed culture
[0160] The seed culture of Clostridium butyricum, Lactobacillus casei, and yeast is the same as described in Example 1.
[0161] Fermentation tank culture
[0162] The seed culture solution of Clostridium butyricum, Lactobacillus casei, and yeast was inserted into 1m at the inoculum of 6%, 12%, and 2% respectively. 3 Fermentation tank, the filling factor is 80% (v / v).
[0163] The composition of the culture medium of the fermenter is: 20g brown sugar, 5g peptone, 3g yeast extract, KH 2 PO 4 1g, (NH 4 ) 2 SO 4 2g, NaCl 3g, CaCO 3 3g, 3g of NaAc, 1000ml of tap water, pH 7.0, and stand anaerobic culture at 35°C for 48h.
[0164] Bacteria collection
[0165] The culture solution was pumped int...
Embodiment 3
[0166] Example 3: Preparation of pure microbe powder
[0167] (1) Bacillus subtilis original bacteria powder
[0168] Mix the wet cells of Bacillus subtilis collected after pure seed cultivation with dry starch in a mass ratio of 1:2, dry at a low temperature of 40-50℃ for 20-24h, crush with a pulverizer, pass through a 40-mesh sieve, and adjust with dry starch The total number of live bacteria of Bacillus subtilis in the original bacteria powder ≥5×10 8 CFU / gram.
[0169] (2) Clostridium butyricum original bacteria powder
[0170] The wet cells of Clostridium butyricum collected after the pure breed culture are mixed with dry starch in a mass ratio of 1:2.5, dried at a low temperature of 40-50℃ for 20-24h, crushed by a pulverizer, passed through a 40-mesh sieve, and dried with a The total number of viable bacteria of Clostridium butyricum in the starch-regulating original bacteria powder ≥5×10 8 CFU / gram.
[0171] (3) Original Lactobacillus casei powder
[0172] The wet cells of ...
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