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Copper-rich bacillus subtilis and cultivating method thereof

A Bacillus subtilis and culture medium technology, applied in the field of copper-rich Bacillus subtilis and its cultivation, can solve the problems of animal and plant product residues, natural ecological environment pollution, water eutrophication, etc., and achieve the promotion of feed utilization , low cost, and the effect of promoting the activity of digestive enzymes

Active Publication Date: 2018-07-31
山东德信生物科技有限公司
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  • Abstract
  • Description
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Problems solved by technology

[0003] As we all know, a certain concentration of copper can promote the growth and development of animals, but when high doses of copper are added to the feed, the utilization rate of copper is low, causing nearly 35% of copper to be excreted from the body, causing pollution to the natural ecological environment and water, especially In modern large-scale livestock and poultry farming, the copper content discharged into the environment is very high, which will cause eutrophication of water bodies and cause residues of animal and plant products, thus endangering the entire ecological environment

Method used

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  • Copper-rich bacillus subtilis and cultivating method thereof
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  • Copper-rich bacillus subtilis and cultivating method thereof

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preparation example Construction

[0033] 4) Preparation of Bacillus subtilis copper: Under the final optimized culture conditions, Bacillus subtilis was fermented for a certain period of time, centrifuged to remove inorganic copper and other components in the medium to obtain a wet sample of Bacillus subtilis copper strain, which was subjected to After drying at 55°C, the content was detected to obtain the Bacillus subtilis copper product.

[0034] The preparation parameters of Bacillus subtilis copper are: ① Medium formula: peptone 10g / L, yeast powder 7g / L, NaCl 3g / L, glucose 7.53g / L, Cu 2+ Concentration 51.2mg / L, pH value 6-8; ②Culture conditions: inoculum size 1-3%, culture temperature 24-36℃, culture time 24-56h, rotation speed 150-250r / min. ③ Target strains were subcultured 18 to 25 times.

[0035] When Cu 2+ When the addition amount of the Bacillus subtilis was 51.2mg / kg, the thalline content of the finally obtained Bacillus subtilis copper was 0.43g / L, and the organic copper content of the thalline wa...

Embodiment 1

[0037] Example 1: Screening of copper-enriched Bacillus subtilis

[0038] 1, the steps of the present invention are as follows:

[0039] 1) Screening of strains: gradually increase Cu in the culture medium 2+ The method of acclimatization is carried out in the form of concentration; after the domesticated strains are continuously cultured for several generations, they are compared with the Bacillus subtilis before domestication in terms of cell morphology, enzyme activity, small peptides, amino acids, etc., to further determine the properties of the domesticated Bacillus subtilis characteristics, and can be stably passaged, and finally obtained the Bacillus subtilis (Bacillus subtilis) NT66 strain that can tolerate high concentrations of copper sulfate (300mg / L), which was preserved in Beichen West Road 1, Chaoyang District, Beijing on January 31, 2018. No. 3, Institute of Microbiology, Chinese Academy of Sciences, General Microbiology Center of China Committee for Culture Co...

Embodiment 2

[0046] Embodiment 2: the establishment of culture method

[0047] 1. Establishment of training methods

[0048] 1) Determination of balance point: according to the growth characteristics of the strain and the 2+ Tolerance ability, with the bacteria content and the copper content of the bacteria as the final measurement index.

[0049] 2) Optimization of culture medium and culture conditions: The culture conditions of Bacillus subtilis copper strain were preliminarily determined by optimizing glucose, pH, culture temperature, rotation speed, inoculum size, passage times, etc. through single factor experiments.

[0050] 3) Response surface analysis: According to the single factor test results, the culture conditions of Bacillus subtilis copper strain were determined by response surface analysis.

[0051] Obtained through single factor preliminary screening, the described medium parameters are: ① medium formula: peptone 10g / L, yeast powder 7g / L, NaCl 3g / L, glucose 5g / L, pH valu...

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Abstract

The invention relates to copper-rich bacillus subtilis and a cultivating method thereof. Bacillus subtilis of probiotics and Cu<2+> are fermented together, and the bacillus subtilis is domesticated bygradually increasing the concentration of CuSO4 to obtain a target bacillus subtilis strain NT66. By domestication of the bacillus subtilis by adopting a method of gradually increasing the concentration of Cu<2+>, the tolerance of the bacillus subtilis can be remarkably improved, and the copper content of thalli is increased. Through the optimization of a culture medium and the fermentation condition, a bacillus subtilis copper product is obtained, so that not only are active substances, such as enzyme, amino acid and small peptides, generated in the thallus fermentation process improved, butalso the copper enrichment amount of bacillus subtilis is increased. The prepared bacillus subtilis copper not only has the advantages of bacillus subtilis, inorganic copper is converted into thallusorganic copper, the utilization rate of animals on trace metal elements is greatly improved, and the biological function of organic copper is exerted well.

Description

technical field [0001] The invention belongs to the technical field of production of livestock and poultry microecological preparations, and in particular relates to a copper-rich Bacillus subtilis and a cultivation method thereof. Background technique [0002] As an intestinal probiotic, Bacillus subtilis can rapidly consume free oxygen in the intestinal tract, create a hypoxic environment in the intestinal tract, promote the growth of beneficial anaerobic bacteria, and produce organic acids such as lactic acid, reduce the pH value of the intestinal tract, and inhibit the growth of pathogenic bacteria. The growth of germs stimulates the growth of immune organs, improves the body's immunity, and increases the growth rate. On the other hand, Bacillus subtilis can adsorb divalent metal cations, and is often used to deal with heavy metal environmental pollution; however, the current application of this strain only considers the effect of intestinal microecology, without taking ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K10/18C12R1/125
CPCA23K10/18C12N1/20C12N1/205C12R2001/125
Inventor 王宝维刘国栋黄燕萍葛文华张名爱岳斌
Owner 山东德信生物科技有限公司
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