A kind of Saccharopolyspora spinosa ds190375 and its fermentation product, bacterial agent, fermentation and screening method and application
A technology of DS190375 and Saccharopolyspora spinosa, which is applied in the field of microorganisms, can solve problems such as unsuccessful industrialization, strain decline, and unstable fermentation production level, and achieve the effects of improving efficiency, increasing screening throughput, and high mutation rate
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Embodiment 1
[0061] Embodiment 1: the salt tolerance experiment of starting strain
[0062] Saccharopolyspora spinosa NRRL18395 purchased from the American Agricultural Culture Collection (NRRL) was used as the starting strain. Prepare a batch of plate culture medium without adding, adding 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0% sodium chloride respectively. Spread the starting strains on the plates respectively, and coat 5 plates for each sodium chloride concentration. After the coated plates are cultured in an incubator at 29°C for 9 days, observe the growth of the colonies on the plates of each group ,As shown in Table 1:
[0063] Table 1 Growth of Saccharopolyspora spinosa NRRL 18395 on salt-containing plates
[0064]
[0065]
[0066] Remarks: +++ indicates a large number of colonies, ++ indicates sparse colonies, + indicates only single-digit colony growth, - indicates no growth.
[0067] It can be seen from the above table 1 that when the concentration of...
Embodiment 2
[0068] Embodiment 2: ARTP mutagenesis screening Saccharopolyspora spinosa high-yield strains
[0069] (1) Incline cultivation of Saccharopolyspora spinosa:
[0070] Using Saccharopolyspora spinosa NRRL18395 purchased from the Agricultural Culture Collection (NRRL) as the starting strain, the starting strain was inoculated on the slant medium, and the slant was cultured in an incubator at 29°C for 9 days to obtain a mature slant.
[0071] (2) Shake flask seed culture of Saccharopolyspora spinosa
[0072] From the mature slant cultivated in step (1), scrape the slant spores and inoculate them into the seed culture medium in the seed shaker flask, insert about 1cm into each bottle 2 The spores on the slant were cultured in the seed bottle at 29° C. on a shaker at 220 rpm for 3 days to obtain a shaker bottle seed liquid.
[0073] (3) Preparation of Saccharopolyspora spinosa protoplast suspension
[0074] Get the shake flask seed liquid cultivated in step (2), add it in a steril...
Embodiment 3
[0092] Example 3: Correlation experiment between high-yield traits and high-salt tolerance traits of mutagenized strains
[0093] In production practice, it was found that the high-yield strains of spinosad were prone to yield degradation during production. As can be seen from Table 2, the high-yield bacterial strains obtained by mutagenesis and screening by the method of the present invention will also have different degrees of decline in production capacity after multiple passages. Some bacterial strains have better stability, and some bacterial strains have more serious degradation. In short, during the amplification process of spinosyn strains under production conditions, the easy degradation of high-yield traits is its own characteristic, which is difficult to overcome. In order to study the relationship between the high-yield traits and salt-tolerant traits of the mutant strains obtained by the method of the present invention, the following experiments were designed.
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