High-flux screening method of nosiheptide active streptomycete high-yield strain

A technology of active Streptomyces and high-yielding strains, which is applied to the field of high-throughput screening of high-yielding strains of Streptomyces spp. active with nosiheptide, can solve the problems of high cost, time-consuming, laborious and other problems, and achieves an accurate and rapid screening method and improves screening efficiency. Effect

Inactive Publication Date: 2015-01-28
ANHUI UNIVERSITY OF TECHNOLOGY AND SCIENCE
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  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the time-consuming, laborious, low efficiency and high cost of current methods for screening high-yield strains of nosiheptide active streptomyces, the present invention can effectively solve the problem by providing a high-throughput screening method for high-yield nosiheptide active streptomyces strains The above existing problems; optimize the fermentation conditions of the 24 deep-well plate at the same time, and verify the correlation between the titer of the 24 deep-well plate and the titer of the shake flask

Method used

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  • High-flux screening method of nosiheptide active streptomycete high-yield strain
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  • High-flux screening method of nosiheptide active streptomycete high-yield strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 24 deep-well plate seed medium loading volume on the impact of Nosiheptide titer

[0029] The initial volume of seed medium was selected as 0.7mL, 1.0mL, 1.3mL, 1.6mL, 1.9mL, 2.2mL, 2.5mL, 2.8mL, 3.1mL respectively, and their effects on the production of nosiheptide in 24 deep-well plates were compared. affect, as a result figure 1 shown. Depend on figure 1 It can be seen that the output of nosiheptide is the highest when the initial liquid loading volume is 2.5mL.

Embodiment 2

[0030] Embodiment 2 24 deep-well plate fermentation medium loading liquid volume on the influence of nosiheptide titer

[0031] According to mass percentage (%), starch 7.0, amylase (0.1% of starch), glucose 0.5, yeast powder 0.2, soybean powder (oil-containing) 4.0, (NH 4 ) 2 SO 4 0.1, KNO 3 0.05, NaCl 0.4, light calcium carbonate 0.4, and sterilized at 115°C for 25 minutes to obtain the fermentation medium.

[0032] Fermentation tests were carried out at 0.7mL, 1.0mL, 1.3mL, 1.6mL, 1.9mL, 2.2mL, 2.5mL, 2.8mL, 3.1mL, 3.4mL of fermentation medium, and the production of Nosiheptide was compared effect, the result is figure 2 shown. When the fermentation medium in the 24-deep-well plate was 1.0 mL, the yield of the 24-deep-well plate was the highest.

Embodiment 3

[0033] Example 3 Effect of fermentation time on 24 deep-well plate culture titer

[0034] Select different fermentation times to measure the output of Nosiheptide, the results are as follows: image 3 shown. Depend on image 3 It can be seen that in the early stage of fermentation, the yield of nosiheptide increases with the prolongation of fermentation time, but its yield decreases after 120 hours of fermentation. Therefore, it is advisable to choose 120 hours of fermentation time.

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Abstract

The invention provides a high-flux screening method of a nosiheptide active streptomycete high-yield strain. A novel ARTP breeding system is utilized to induce mutation of nosiheptide active streptomycete; a response surface process is utilized to optimize the 24-deep-hole plate fermentation conditions; and a 24-deep-hole plate culture process is utilized to establish the high-flux mutant screening method, thereby obtaining the high / stable-yield nosiheptide mutant strain. After verifying the relevance between the 24-deep-hole plate titer and shake flask titer, the screening method is very accurate and quick. Compared with the traditional shake flask fermentation screening process, the 24-deep-hole plate process can obviously enhance the screening efficiency of the nosiheptide high-yield strain.

Description

technical field [0001] The invention belongs to the field of microbial mutation breeding and fermentation engineering, and in particular relates to a high-throughput screening method for nosipeptide-active Streptomyces high-yield strains. Background technique [0002] Nosiheptide is a sulfur-containing polypeptide antibiotic, also known as Nosiheptide, which is mainly produced by Streptomyces actives. It has the advantages of promoting animal growth, improving feed utilization, enhancing animal disease resistance, and has no residue in animals, safe for humans and animals, and less environmental pollution. It is an ideal substitute for antibiotics for feeding. [0003] At present, in the European Union, Japan, Taiwan and other regions, nosiheptide is added to the feed as the only feed antibiotic. Domestic researchers have done a lot of research on the breeding of nosiheptide-active streptomyces strains and the optimization of fermentation process, which has greatly improved...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12N15/01C12R1/46
Inventor 薛正莲周扬秦艳飞赵世光王洲
Owner ANHUI UNIVERSITY OF TECHNOLOGY AND SCIENCE
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