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Screening method of avilamycin high-producing strain

A technology of avelamycin and high-yield strains, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problem of poor mutagenesis treatment, poor fermentation level of strains, uneven mycelium dispersion, etc. problems, to achieve good industrial application potential, high content of effective components, and stable fermentation effects

Inactive Publication Date: 2017-09-15
GUANGDONG RONGDA BIOENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Around 2005, there were reports on the breeding of avelamycin strains and the research on fermentation conditions in China. However, the relevant research was still in its infancy, and the level of production was low, and there were many impurities in the fermentation, which further affected the follow-up. Separation and purification bring difficulties and increase production costs. The reason is that the fermentation level of the strain is not good
[0005] S.viridochromogene, as a type of actinomycetes, grows in the form of mycelium and reproduces as spores, and has a cell wall. Direct mutagenesis treatment often has poor treatment effects, and the effect of mutagenesis treatment will be caused by uneven dispersion of mycelia. Not good. In addition, the special structure of spores often makes them insensitive to mutagen, which will cause a lot of false positive effects

Method used

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  • Screening method of avilamycin high-producing strain
  • Screening method of avilamycin high-producing strain
  • Screening method of avilamycin high-producing strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The preparation method of embodiment 1 medium and buffer

[0035] Slant and solid plate medium (g L -1 ): soluble starch 20.0, MgSO 4 0.35, KNO 3 1.2,K 2 HPO 4 0.25, KH 2 PO 4 0.55, FeSO 4 0.01, NaCl 0.5g / L, agar powder 20.0, NaOH to adjust the pH to 7.2-7.5, sterilize at 121°C for 20min, and set aside.

[0036] Seed medium (g L -1 ): Soluble starch 20.0, soybean meal extract powder 8.0, soybean peptone 5.0, D-xylose 7.5, CaCO 3 0.5, FeSO 4 0.01, MgSO 4 0.5, MnCl 2 0.5, adjust the pH to 7.2-7.5 with NaOH, sterilize at 121°C for 25 minutes, and set aside.

[0037] Fermentation medium (g L -1 ): Soluble starch 20.0, soybean meal extract powder 8.0, soybean peptone 5.0, D-xylose 7.5, L-valine 3.0, CaCO 3 0.5, FeSO 4 0.01, MgSO 4 0.5, MnCl 2 0.5, adjust the pH to 7.2-7.5 with NaOH, sterilize at 121°C for 25 minutes, and set aside.

[0038] Regeneration medium (g L -1 ): soluble starch 20.0, MgSO 4 0.35, KNO 3 1.2,K 2 HPO 4 0.25, KH 2 P...

Embodiment 2

[0042] Embodiment 2 A kind of screening method of avelamycin high-yielding bacterial strain

[0043] The screening method of described avelamycin high-yielding bacterial strain, comprises the following steps:

[0044] S1. Take a slant of Streptomyces viridans AVL4 that has been activated and cultivated, as the starting strain, add 5mL sterile normal saline to elute, prepare the spore suspension, and transfer it into the culture medium containing 5% glycine according to the inoculation amount of 5%. In a 250mL Erlenmeyer shaker flask with baffles filled with 50mL seed medium, continue to cultivate at 28°C and 220rpm for 24h. After the cultivation, centrifuge the bacterial solution at 6000rpm at 4°C for 10min, wash twice with phosphate hypertonic buffer, and discard the supernatant , to obtain the starting mycelia prepared by protoplasts;

[0045] S2. Using SMM solution as an osmotic pressure stabilizer, add a final concentration of 12 mg mL to the starting mycelium tube prepar...

Embodiment 3

[0050] Embodiment 3 A kind of screening method of avelamycin high-yielding bacterial strain

[0051] The screening method of described avelamycin high-yielding bacterial strain, comprises the following steps:

[0052] S1. Take a slant of Streptomyces viridans AVL4 that has been activated and cultivated, as the starting strain, add 4mL sterile normal saline to elute, prepare the spore suspension, and transfer it to the container containing 5% glycine according to the inoculation amount of 4%. In a 250mL Erlenmeyer shaker flask with baffles containing 50mL seed medium, continue to cultivate at 27°C and 210rpm for 24h. After the cultivation, centrifuge the bacterial solution at 5000rpm at 4°C for 5min, wash twice with phosphate hypertonic buffer, and discard the supernatant. Obtain the starting mycelium of protoplast preparation;

[0053] S2. Using SMM solution as an osmotic pressure stabilizer, add a final concentration of 9 mg mL to the starting mycelium tube prepared in step ...

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Abstract

The invention relates to the technical field of screening of microbial strains, and in particular to a screening method of an avilamycin high-producing strain. The method comprises the following steps: preparing streptomyces viridocbromogene protoplast; inducing mutation of the protoplast by virtue of an ARTR system; implementing preliminary screening in accordance with morphological characteristics and colony bacteriostatic biological value of the induced strain; and then, implementing shaking flask secondary screening by taking avilamycin bacteriostatic value and avilamycin A component content as indexes; and the strain, obtained from screening, has the advantages of being high in fermenting stability and high in avilamycin yield, and being high in content of effective components. According to the screening method provided by the invention, cost of preparing related products is reduced, and an excellent potential of industrial application is shown.

Description

technical field [0001] The invention relates to the technical field of screening microbial strains, in particular to a screening method for high-yielding avelamycin strains. Background technique [0002] Avilamycin (Avilamycin), also known as bemcillin, can inhibit the synthesis of related proteins by binding to the ribosomes of heterogeneous Gram-positive bacteria in the intestinal tract, thereby maintaining the ecological balance of the overall probiotic flora, and at the same time It can also inhibit intestinal microbial lactic acid production, thereby reducing intestinal peristalsis, delaying the residence time of nutrients in the intestinal tract of animals, promoting digestion and regulating metabolism, thereby promoting animal growth. Due to its unique structure, high safety, easy degradation, almost no residue, growth promotion and disease resistance, avilamycin is widely used in animal feed, especially pig and broiler feed. [0003] Streptomyces viridochromogene is...

Claims

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Application Information

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IPC IPC(8): C12N13/00C12N1/20C12Q1/18G01N30/02C12R1/465
CPCC12N13/00C12N1/20C12Q1/18G01N30/02
Inventor 蒋顺进刘宗新郑雪媚黄炜乾唐谢芳
Owner GUANGDONG RONGDA BIOENG CO LTD
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