A Wuyiencin high-yielding strain, fermentation method and application

A technology of Wuyiencin and high-yield strains, applied in the field of bioengineering, can solve the problems of strong mutation randomness, heavy screening workload, and unstable strains

Active Publication Date: 2022-05-03
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mutant strains obtained by ARTP technology have a certain degree of randomness, and the workload of screening after mutation is large, the mutation is highly random, and problems such as back mutation and strain instability are prone to occur. High-yielding strains are also unknown

Method used

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  • A Wuyiencin high-yielding strain, fermentation method and application
  • A Wuyiencin high-yielding strain, fermentation method and application
  • A Wuyiencin high-yielding strain, fermentation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] 1. Cultivation of Wuyiencin-producing strains

[0055] Use a sterile toothpick to transfer the Wuyiencin-producing strain-Streptomyces nonhygroscopicus var. Wuyi CK15 to MS medium, spread the plate, and culture it at 28°C.

[0056] MS medium components are: mannitol 20g / L, soybean powder 20g / L, agar powder 20g / L, taking the preparation of 1L as an example, first weigh the components of the medium, then add water to make up to 1L.

[0057] 2. Fermentation and cultivation of Wuyiencin-producing strains

[0058] Cut 2 cm of CK15 from MS medium with a scalpel 2 Inoculate into a Erlenmeyer flask equipped with 200mL fermentation medium, and culture at 28°C for 64h on a shaker at 220rpm.

[0059] The components of the fermentation medium are: corn flour 20g / L, soybean flour 25g / L, glucose 25g / L, calcium carbonate 4g / L.

[0060] 3. Mutation of strains

[0061] a) Streptomyces nonhygroscopicus var. Wuyi CK15 was inoculated into a MS plate with a diameter of 9 cm and cultured...

Embodiment 2

[0084] 1. Fermentation culture in 5L fermenter of Wuyiencin producing strain

[0085] (1) Bacteria culture: inoculate CK15 and M28 on 10 MS plates with a diameter of 9 cm respectively, and culture 5 plates for each of the two strains at 28°C for 7 days. There is no significant difference in the colony morphology between the two strains (such as Figure 4), the aerial hyphae are gray, with white secondary hyphae in between; the basal hyphae are light yellow or light yellow, producing light yellow, yellowish or yellow soluble pigments. Pick a well-growth plate respectively, and then use 8mL sterile water to elute the spores into 15mL sterile corning tubes;

[0086] The composition of MS medium is the same as that in Example 1.

[0087] (2) Fermentation culture: Inoculate all the eluted spore liquids of the above two bacterial strains (each bacterial strain is eluted with 8mL sterile water, and finally eluted to obtain 5-6mL spore liquid) into a 5L fermenter, with 3L Medium, at...

Embodiment 3

[0091] The 500L fermenter fermentation culture of embodiment 3 Wuyiencin producing bacterial strains

[0092] (1) Strain culture: inoculate CK15 and M28 on 20 MS plates with a diameter of 9 cm respectively, and culture 10 plates for each of the two strains at 28°C for 7 days, and then use 60 mL of sterile water to elute the spores to 200 mL in a sterile Erlenmeyer flask;

[0093] The composition of MS medium is the same as that in Example 1.

[0094] (2) Seed liquid culture: Inoculate the above spore liquid into a 50L fermenter with 30L medium inside, at a temperature of 28°C, a speed of 200rpm, an air flow ratio of 1:0.5, and a tank pressure of 0.1kg / cm 2 Cultivate 24h under the condition of obtaining seed liquid;

[0095] The medium components are: glucose 10g / L, peptone 5g / L, yeast powder 3g / L, sucrose 260 g / L.

[0096] (3) Fermentation tank culture: Transfer the vigorously growing primary seed liquid into a 500L fermenter tank with 300L fermentation medium, at a tempera...

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Abstract

The invention relates to a Wuyiencin high-yielding strain, a fermentation method and application. The preservation number of the strain is CGMCC No.19060. The fermentation method of the bacterial strain comprises the following steps: inoculating the bacterial strain into a culture medium, and fermenting and cultivating. The bacterial strain can be used to prepare Wuyiencin, and the fermentation liquid of the bacterial strain has good antibacterial effect on various pathogenic bacteria. The strain of the invention has the advantages of rapid growth, high spore production, high yield of Wuyiencin, high potency and stability, and the like.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a Wuyiencin high-yielding strain, fermentation method and application. Background technique [0002] Wuyimectin is a low-toxicity, high-efficiency, broad-spectrum, and environmentally-friendly new agricultural antibiotic with completely independent intellectual property rights in my country (the chemical structural formula is shown in formula 1). The fungal diseases of many kinds of crops, such as fungal diseases, have obvious control effects, and have been widely used in agricultural production, creating considerable economic, social and ecological benefits. However, the current production is facing the problems of strain degradation, low fermentation titer and high production cost, which restricts the industrialization of Wuyiencin. [0003] [0004] High-yield strain breeding usually adopts physical mutagenesis (ultraviolet, gamma-ray, laser, etc.), chemi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N13/00C12N15/01C12P19/38A01N43/54A01N63/28A01P3/00C12R1/465
CPCC12N1/20C12N13/00C12N15/01C12P19/385A01N43/54A01N63/28
Inventor 施李鸣葛蓓孛张克诚韦秋合杨淼泠张维
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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