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Saccharopolyspora spinosa DS190375 as well as fermentation product, microbial inoculum, fermentation and screening method and application thereof

A technology of DS190375, Saccharopolyspora spinosa, applied in the field of microorganisms, can solve problems such as unsuccessful industrialization, unstable fermentation production level, and obstacles to the successful industrialization of spinosad

Active Publication Date: 2020-10-30
NEW FOUNDER HLDG DEV LLC +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, many scientific research institutions have successfully used methods such as mutagenesis screening, protoplast fusion, and genetic engineering to improve the fermentation units of spinosad strains, but none of them have been successfully industrialized. One of the important reasons is that the high yield of spinosad Bacterial strains are very prone to decline in the production process, resulting in very unstable fermentation production levels and hindering the successful industrialization of spinosads

Method used

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  • Saccharopolyspora spinosa DS190375 as well as fermentation product, microbial inoculum, fermentation and screening method and application thereof
  • Saccharopolyspora spinosa DS190375 as well as fermentation product, microbial inoculum, fermentation and screening method and application thereof
  • Saccharopolyspora spinosa DS190375 as well as fermentation product, microbial inoculum, fermentation and screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to the technical field of microorganisms, and particularly discloses saccharopolyspora spinosa DS190375 as well as a fermentation product, a microbial inoculum, a fermentation and screening method and an application of saccharopolyspora spinosa DS190375. The preservation number of the saccharopolyspora spinosa DS190375 disclosed by the invention is CGMCC No. 18681. The fermentation method comprises the following steps: rejuvenating and screening the strain in a high-salt plate culture medium, and fermenting. The invention also provides a screening method of the saccharopolyspora spinosa with high spinosad yield, which comprises the following steps: screening to obtain the saccharopolyspora spinosa with high salt tolerance, and screening the obtained saccharopolysporaspinosa with high salt tolerance to obtain the strain with high spinosad yield. The saccharopolyspora spinosa DS190375 disclosed by the invention is resistant to high salt and high in spinosad yield.The fermentation method avoids the problem of unstable spinosad yield caused by strain performance degradation.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Saccharopolyspora spinosa DS190375 and its fermentation product, bacterial agent, fermentation and screening method and application. Background technique [0002] Spinosad is a kind of macrolide biologically active substance produced by Saccharopolyspora spinosa, which has a high-efficiency killing effect on most of the Lepidoptera and Diptera agricultural pests. Other agricultural pests also have a certain control effect. The key is that spinosad is very safe for people and the environment, and it is biodegradable. It is an ideal "green pesticide". [0003] At present, the key factors restricting the large-scale industrialization of spinosyn are its complicated fermentation process, low fermentation unit and high production cost. Therefore, increasing the fermentation unit of spinosad and reducing its production cost are the current research focus. In recent years, ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/01C12N13/00C12P19/62C12R1/01
CPCC12N1/20C12N15/01C12N13/00C12P19/62C12R2001/01C12N1/205
Inventor 赵子刚刘露金明义张葵
Owner NEW FOUNDER HLDG DEV LLC
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