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Bacillus subtilis and application thereof

A Bacillus subtilis, single colony technology, applied in the field of microorganisms, can solve the problems of proteases not having high temperature resistance, low protease production activity, poor application effect, etc., to improve the intestinal microbial environment, fully digest food, and bitter peptides The effect of low content

Active Publication Date: 2019-02-05
广州大峰收技术服务有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

Inventors have developed new types of bacilli called Bifidobactrium sp., which make up about 7% of all diarrhea cases caused by gastrointesting harmful organisms such as Escherichia coli (ECO) among others. These bacterias help break down certain substances like fiber from cow's milk while also being able to stimulate beneficial gut flora growth for improved nutritional value. They work well at temperatures between 0°C and 150 degrees Celsium but they cannot effectively decompose them due to their highly active ingredients. By adding these specialized bacils together into feeds containing other important components, we aimed towards creating stronger and tasteier versions of those added supplements.

Problems solved by technology

The technical problem addressed in this patented text relates to finding better ways for producing Bacilli with improved properties like antibiotics or other beneficial substances from their natural sources while maintaining their effectiveness at higher temperatures than current methods due to poor cellular entry mechanisms during feed ingestion (gastrointestinal).

Method used

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  • Bacillus subtilis and application thereof
  • Bacillus subtilis and application thereof
  • Bacillus subtilis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1, Screening of Bacillus subtilis (Bacillus subtilis) CDMCC NO.60294

[0040] 1. Primary screening

[0041] (1) Reagents used:

[0042] The preparation method of the physiological saline is: 0.85wt% sodium chloride, and it is used after autoclaving.

[0043] The preparation method of the artificial gastric juice is: 1%wt pepsin, 0.85%wt sodium chloride, adjust the pH value to 1.5 with HCl, and use a 0.22μm bacterial filter to filter and sterilize for future use.

[0044] The preparation method of the artificial bile salt is as follows: adding 0.3wt% pig bile salt (analytical pure) to the liquid broth, and autoclaving for subsequent use.

[0045] The preparation method of LB solid medium is: yeast extract 5g, tryptone 10g, sodium chloride 10g, agar 15g, add water to 1000mL, adjust pH to 7.0 with NaOH, autoclave at 121°C for 20min, cool to 50°C, pour flat.

[0046] (2) method:

[0047] Pig manure, chicken manure, cow manure, tempeh, rotten rice straw, pig fa...

Embodiment 2

[0062] Embodiment 2, identification of Bacillus subtilis (Bacillus subtilis) CDMCC NO.60294

[0063] (1) Preliminary identification of Bacillus subtilis Z-2 (DFS01, CDMCC NO.60294)

[0064] The colony morphology of the screened bacteria Z-2 (DFS01, CDMCC NO.60294) was observed, and the morphological characteristics of the bacteria were observed after Gram staining. The surface of the colony is rough and wrinkled, opaque, dirty white, the colony is round, crater-shaped, with jagged edges, Gram-positive bacteria; the spores are oval to columnar, located in the center of the cell or slightly biased, after spore formation Bacteria do not expand. It was preliminarily speculated to be a strain of the genus Bacillus.

[0065] (2) Molecular identification of Bacillus subtilis Z-2 (DFS01, CDMCC NO.60294)

[0066] Use the bacterial universal primers to carry out PCR identification on the screened bacterial strains: extract the template DNA according to the operating instructions of...

Embodiment 3

[0075] Embodiment 3, preparation Bacillus subtilis (Bacillus subtilis) DFS01 liquid agent-L1

[0076] (1) Primary slant culture of Bacillus subtilis:

[0077] Inoculate a ring of bacteria solution in the glycerol storage tube of Bacillus subtilis (Bacillus subtilis) Z-2 (DFS01, CDMCC NO.60294) on the first-grade slant medium, and cultivate it at 37°C for 25 hours to obtain the first-grade Bacillus subtilis Bacillus single colony; the described primary slant medium is a mixed solution (g / L) configured in the following proportions: soybean peptone 6.0, yeast extract 6.0, sodium chloride 1.0, glucose 4.0, agar 20, add water to 1000mL, pH 7.0.

[0078] (2) Secondary liquid culture of Bacillus subtilis:

[0079] The first-level Bacillus subtilis single bacterium colony that step (1) cultivates is inoculated in the secondary culture fluid, and liquid volume 250mL / 1000mL cultivates 25 hours under 35 ℃, 150rpm conditions, obtains the second-level Bacillus subtilis seed liquid; Th...

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Abstract

The invention relates to bacillus subtilis DFS01 and application thereof. The collection number of the strain is CDMCC NO. 60294. Compared with the existing known bacillus subtilis strain, the bacillus subtilis DFS01 is remarkably characterized in that (1) the strain has high enzyme production activity, the produced proteases comprise keratinase, carboxypeptidase and aminopeptidase which can withstand the temperature of 100 DEG C compared with proteases produced by ordinary bacillus subtilis strains, the content of bitter peptides produced by decomposing the proteases is low, and a fermentation product has very strong flavor and very high livestock palatability; (2) the strain has the characteristic of high intestinal cell adhesion capability, and the adhesion rate is up to 62.78 percent.

Description

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Claims

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Application Information

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Owner 广州大峰收技术服务有限公司
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