Enterococcus faecium, culture medium thereof for high-density solid-state fermentation, and high-density solid-state fermentation method
A technology of Enterococcus faecium and culture medium, which is applied in the field of Enterococcus faecium and its high-density solid-state fermentation medium, can solve the problems of easy feedback control, easy inactivation of strains, high cost, etc., and achieve good economic benefits and live bacteria The effect of high concentration and low production cost
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Embodiment 1
[0040] Example 1 Isolation, screening, identification and preservation of Enterococcus faecium
[0041] 1.1 Isolation of strains
[0042] Take dry cheese samples, grind them, and dilute them 10, 100, and 1000 times with sterile water, respectively, take 200ul and spread them on MRS solid medium, culture them at 37°C for 48 hours, pick colonies, streak MRS medium, and After 4 times of purification, a single colony was picked and cultured in MRS broth, 20% glycerol, and stored at -70°C for future use. After Gram staining, catalase test, nitrate reduction test and 16SrDNA sequencing.
[0043] 1.2 Identification of strain morphological characteristics
[0044] Take a single colony of the purified strain (GLM105), transfer it to MRS solid medium (agar), and culture it in a constant temperature incubator at 37°C for 24h, 36h, and 48h, and observe the size, color, edge, protrusion, and Smoothness, viscosity, transparency and other characteristics. The results showed that strain G...
Embodiment 2
[0087] Embodiment 2 A kind of culture medium of high-density solid-state fermentation Enterococcus faecium
[0088] The formula of the solid-state fermentation medium is: 22% wheat bran, 22% soybean meal, 4% sugar cane molasses, 3% activated carbon, 5% calcium carbonate, 4% β-cyclodextrin, and 40% water content, and the percentages are all by mass . (except that the formula is, the formula of the solid-state fermentation medium can be: wheat bran 15%~25%, soybean meal 19%~29%, sugarcane molasses 3%~5%, activated carbon 2%~4%, calcium carbonate 2% ~6%, β-cyclodextrin 2% ~ 6%, water content 40% ~ 50%, the percentages are mass percentages.)
[0089] The preparation method of the above-mentioned medium: first configure the basic medium (wheat bran, soybean meal and water) according to the conventional method, then add sugarcane molasses, activated carbon, calcium carbonate, β-cyclodextrin, stir well, and autoclave at 115°C for 15 minutes , and cool down. Wheat bran and soybean ...
Embodiment 3
[0090] Embodiment 3 A kind of method of high-density solid-state fermentation culture Enterococcus faecium
[0091] 1) Preparation of strains and their seed solution
[0092] The Enterococcus faecium GLM105 strain was activated on the MRS agar plate, and a single colony was picked and inoculated into the MRS liquid test tube, cultured at 37°C for 24 hours, then transferred to the MRS liquid medium with 2% v / v inoculum, and cultured for 24 hours as the seed solution.
[0093] The main components of MRS medium are peptone 10.0g, beef extract 10.0g, yeast extract 5.0g, diammonium hydrogen citrate 2.0g, glucose 20.0g, Tween 801.0mL, sodium acetate 5.0g, dipotassium hydrogen phosphate 2.0g, sulfuric acid Magnesium 0.58g, manganese sulfate 0.25g, distilled water 1000mL, pH6.2~6.6. The optimum pH is 6.0, the optimum temperature is 37°C, facultative anaerobic.
[0094] 2) Vaccination
[0095] With 3% v / v inoculation amount, the prepared seed liquid is inoculated into the substratum...
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