Composite microbial preparation as well as preparation method and application thereof
A technology of compound microorganisms and microorganisms, which is applied in the field of compound microbial preparations and its preparation, can solve problems such as complex and cumbersome formulas, cumbersome preparation processes, and complex ratios of microbial preparations, and achieve the effects of reducing production costs and improving fermentation efficiency
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Embodiment 1
[0057] Example 1 Screening of Bacillus subtilis
[0058] The straws are kneaded and crushed to 10-20 mm, water is added to adjust the water content to 60%-70%, and natural fermentation is carried out for 3 days. Weigh 5 g of naturally fermented straw, add 45 mL of 0.85% physiological saline, and shake for 30 minutes.
[0059] Enrichment and separation of Bacillus: Take 5 mL of supernatant, add it to a sterile test tube, and bath in water at 80°C for 20 minutes. Inoculate 10% of the inoculum into LB liquid medium, and enrich in a shaker at 180r / min and 30°C for 24h. Perform a gradient dilution of the enriched culture solution, select 10 -4 , 10 -5 , 10 -6 The gradient was spread on the LB agar medium, 3 parallels for each dilution, cultured at 37°C for 24h to obtain a single colony, and the single bacteria were picked and stored in the LB slant medium.
[0060] Screening and identification of cellulase-producing bacillus: Using the plate streaking method, select strains from the inc...
Embodiment 2
[0061] Example 2 Screening of Lactobacillus
[0062] The straws are kneaded and crushed to 10-20 mm, water is added to adjust the water content to 60%-70%, and natural fermentation is carried out for 3 days. Weigh 5 g of naturally fermented straw, add 45 mL of 0.85% physiological saline, and shake for 30 minutes.
[0063] Enrichment and separation of lactic acid bacteria: draw 5 mL of supernatant, add it to a sterile test tube, and bath in a water bath at 80°C for 20 minutes. It was inoculated into MRS liquid medium at 10% inoculum, and enriched on a shaker at 180r / min and 30℃ for 24h. Perform a gradient dilution of the enriched culture solution, select 10 -4 , 10 -5 , 10 -6 The gradient was spread on MRS agar medium, 3 parallels for each dilution, and anaerobic culture at 37°C for 24h to obtain a single colony. According to the difference of colony morphology, various typical colonies were picked, and four-zone streaking was performed on another MRS solid plate, and cultured ana...
Embodiment 3
[0066] Example 3 Screening of Saccharomyces cerevisiae
[0067] The straws are kneaded and crushed to 10-20 mm, water is added to adjust the water content to 60%-70%, and natural fermentation is carried out for 3 days. Weigh 5 g of naturally fermented straw, add 45 mL of 0.85% physiological saline, and shake for 30 minutes.
[0068] Enrichment and separation of yeast: Take 5 mL of supernatant, add it to a sterile test tube, and bath at 80°C for 20 minutes. It was inoculated into YPD liquid medium at 10% inoculum amount, and enriched on a shaker at 180r / min and 30°C for 24h. Perform a gradient dilution of the enriched culture solution, select 10 -1 , 10 -2 , 10 -3 The gradient was spread on YPD agar medium, 3 parallel for each dilution, and cultured at 30°C for 24h to obtain a single colony. According to the difference of colony morphology, various typical colonies were picked, and four-zone streaking was performed on another YPD solid plate, and cultured anaerobic at 30°C for 24h...
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