Bacillus licheniformis for feed and application of bacillus licheniformis
A technology of Bacillus licheniformis and Escherichia coli, applied in the field of microbiology, can solve the problems of less research and application
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Embodiment 1
[0026] Example 1 Isolation and identification of Bacillus licheniformis (Bacillus licheniformis) M5
[0027] 1. Isolation of strain M5
[0028] 1. Isolation and cultivation of strains
[0029] Take 1g of the soil sample near the pig breeding base of the West Campus of China Agricultural University in Haidian District, Beijing, and put it into a test tube with 9ml of normal saline, vortex and shake it to make a 1:10 dilution, and then take the dilution to increase by 10 times Dilute, and then select 1ml of each of 3 appropriate gradient dilutions and spread them on the LB medium. Cultivate at 37°C for 48-72 hours, observe and record the colony shape, pick a single colony with good growth, and separate and purify by streaking.
[0030] 2. UV mutagenesis and screening of strains
[0031] Pour the sterilized LB medium into a petri dish, and after solidification, take the bacterial suspension obtained in step 1 and spread it on a plate. Control the number of colonies in each pet...
Embodiment 2
[0045] Stress resistance detection of embodiment 2 bacillus licheniformis M5
[0046] 1. Heat resistance test
[0047] The Bacillus licheniformis M5 (CGMCC No.15453) bacterial solution was treated in an 80°C water bath for 10 min, 20 min, and 30 min, respectively, and each treatment was repeated three times. After the treatment, the number of viable bacteria was determined by pouring method.
[0048] The survival rate of Bacillus licheniformis M5 CGMCC No.15453 can reach 95% after being treated at 80°C for 10 minutes, and the survival rate still reaches 77% after 30 minutes.
[0049] 2. Acid resistance test
[0050] Bacillus licheniformis M5 was inoculated into LB medium with pH values of 2.0, 3.0, and 4.0, respectively, and the number of viable bacteria was determined by plate pouring method at 0h, 2h, 4h, and 6h, respectively.
[0051] The activity of Bacillus licheniformis M5 CGMCC No.15453 was less affected in the medium of pH 3.0 and 4.0, and the number of viable bact...
Embodiment 3
[0054] The growth curve determination of embodiment 3 bacillus licheniformis M5
[0055] The growth curve represents the dynamic changes in the whole process of bacteria growing and multiplying in a new suitable environment until aging and death. Bacillus licheniformis M5 CGMCC No.15453 was inoculated into LB medium at an inoculum amount of 1% (v / v), and cultivated at 37°C for 36 hours. The LB medium without bacterial solution was used as a blank control, and every 2 Measure the OD600 value at -6 hours. The experiment was repeated three times, the results were averaged, the data were recorded and the growth curve was drawn. Such as Figure 5 As shown, in 5-28 hours, Bacillus licheniformis M5 was in the logarithmic growth phase, and the reproduction rate was higher. The number of Bacillus licheniformis M5 tends to stabilize at 28-30 hours. After 30 hours, the growth rate decline phase was entered.
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