A kind of compound probiotics and preparation method thereof
A technology of composite microecology and preparation, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as improper post-processing, overfilling, and small environmental adaptability of strains, so as to achieve good product stability and improve Strong immune activity and anti-pollution ability
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Embodiment 1
[0034] The acquisition of embodiment 1 symbiotic strain combination
[0035] All the following strains are considered as candidate strains for the research of microecological preparations: (1) Newly preserved strains with excellent functions in our laboratory: Bacillus subtilis ( Bacillus subtilis ) CGMCC No.9356 and Enterococcus faecium ( Enterococcus Faecium ) CGMCC No.9134; (2) The laboratory has preserved excellent strains including: Bacillus licheniformis (Bacillus licheniformis)CGMCC No.7030; Saccharomyces cerevisiae ( Saccharomyces cerevisiae ) CGMCCNo.6921; Lactobacillus pumilus ( Lactobacillus brevis ) CGMCC No.4756; Lactobacillus johnsonii ( Lactobacillus johnsonii ) CGMCC No.4926, Lactobacillus acidophilus (Lactobacillus acidophilus) CGMCC No.6499; Pediococcus lactis ( Pediococcus acidilactic) CGMCC No.6500; (3) Commercially available strains: the following strains were isolated from commercially available single-strain powder: 1 strain of Lactococcu...
Embodiment 2
[0038] Embodiment 2 9 strains of bacteria liquid mixed culture
[0039] The symbiotic bacterial strain combination of 9 strains was screened, and after the 9 strains were cultured in the same medium YH, 1 mL of the seed solution of each strain with the same concentration was taken, and mixed into a certain volume of liquid medium YH for mixed culture. The fermentation temperature 33°C, shaker speed 100rpm, initial pH value 7.0, continuous culture for 4 days, record the number of live bacteria on 0, 3 and 4 days, the results are shown in Table 1: after 4 days of mixed culture of 9 strains, the total concentration of bacillus reached 1.4 ×10 12 CFU / mL or more, the total concentration of lactic acid bacteria is 6.08×10 11 CFU / mL or more, the yeast concentration is 6.52×10 8 More than CFU / mL, it shows that the 9 strains grow well together.
[0040] Table 1
[0041]
Embodiment 3
[0042] Example 3 Preparation of 9 strains of bacterial seed liquid
[0043] (1) Preparation of seed solution of 4 strains of Bacillus
[0044] Bacillus subtilis H8-1, the preservation number is CGMCC No.9356; Bacillus megaterium; Bacillus licheniformis (Bacillus licheniformis) CGMCC No.5094; single colonies of Bacillus amyloliquefaciens CGMCC No.7030 were inoculated into 250mL Erlenmeyer flasks containing 100ml of MNB medium, the shaker speed was 180rpm, and cultured in a constant temperature shaker at 33°C for 16h; the prepared The first-grade seed solution, with an OD600 of about 0.9, was inoculated into a 1L Erlenmeyer flask containing 500mL of MNB medium according to the inoculum amount of 1.0%, and cultivated in a constant temperature shaker at 33°C for 12h, with a shaker speed of 180rpm, and an OD600 of about 0.8 ;
[0045] (2) Preparation of 4 strains of lactic acid bacteria seed solution
[0046] Enterococcus faecium, the preservation number is CGMCC No.9134; Lacto...
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