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Avirulent Bacillus cereus strain pBC-1 and application thereof

A Bacillus cereus, pbc-1 technology, applied in applications, bacteria, disinfectants, etc., can solve problems such as human health and public health hazards

Active Publication Date: 2016-07-13
朱奎
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although Bacillus cereus has the above-mentioned value at present, due to various reasons, the probiotics of Bacillus cereus currently on the market express at least one bacterial toxin, which poses a great hazard to human health and public health

Method used

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  • Avirulent Bacillus cereus strain pBC-1 and application thereof
  • Avirulent Bacillus cereus strain pBC-1 and application thereof
  • Avirulent Bacillus cereus strain pBC-1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Isolation and identification of Bacillus cereus strain pBC-1

[0023] Take 5g of soil from Jiangsu Wuyi Farm, dissolve it in 10mL of sterile phosphate buffer (pH7.2, 0.1mM), and take 0.5mL to spread on commercial Bacillus cereus selective medium (PEMBA, Thermo) , cultivated at 32°C for 18h. The positive colonies were selected and cultured on the Columbia sheep blood agar plate, and a single colony was selected to be identified as Bacillus cereus by phenotype analysis, biochemical level identification and 16SrRNA sequence comparison. The colony morphology was as follows: figure 1 As shown, it is light gray, opaque, approximately round, with a smooth surface.

Embodiment 2

[0024] Example 2 The cultivation and toxicity detection of Bacillus cereus strain pBC-1

[0025] (1) Bacterial culture

[0026] a. Casein hydrolysis yeast medium (CGY) medium: casein hydrolyzate 20g, yeast extract 6g, ammonium sulfate 2g, dipotassium hydrogen phosphate 14g, potassium dihydrogen phosphate 6g, sodium citrate 1g, magnesium sulfate 2g, Add water to 1L and autoclave for later use.

[0027] b. 10% glucose solution: dissolve 10g of glucose in 100mL of distilled water, sterilize with a 0.22μm membrane filter, store at 4°C for later use.

[0028] c. Configuration of sugar-containing casein hydrolytic yeast culture medium (CGY): Add appropriate amount of 10% glucose solution to the casein hydrolytic yeast culture medium until the final concentration of glucose is 1%, and it is prepared and used immediately.

[0029] d. 10% skim milk medium: take 10g of skim milk, add 100mL of distilled water, and prepare it for immediate use.

[0030] The specific culture method of B...

Embodiment 3

[0046] Preparation and application effect of embodiment 3 bacillus cereus spore feed additive

[0047] (1) Preparation of Bacillus cereus spore preparation

[0048] A. Blood plate culture: inoculate the Bacillus cereus strain pBC-1 preserved at low temperature on a Columbia sheep blood agar plate, and after cultivating at 32°C for 24 hours, pick a single colony and repeatedly culture it once on a Columbia sheep blood agar plate;

[0049] B. Preparation of seed solution: make 1×10 strains cultured in step A with sterilized PBS (0.1mM, pH7.2) 8 CFU / mL bacterial suspension, take 1 mL bacterial suspension inoculum and inoculate in 20 mL conventional CGY liquid medium containing 1% glucose, 250 rpm, 32 ° C water bath for 6 h, to obtain seed liquid;

[0050] C. Preparation of spores: Inoculate the 1% dilution of the seed solution in step B in the soybean casein agar medium, cultivate at 32°C for 7 days, and obtain the crude spore extract of Bacillus cereus pBC-1.

[0051] D. Prepa...

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Abstract

The invention discloses an avirulent Bacillus cereus strain pBC-1, which is preserved in China General Microbiological Culture Collection Center (CGMCC, in Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard No.1, Beicheng West Road, Zhaoyang District, Beijing City, China, 100101), with an accession number of CGMCC No. 9535. Identification results show that the sequence of the strain pBC-1 is Bacillus cereus. Results of analysis at the levels of genes, protein and cells, the strain pBC-1 provided by the invention does not generate any toxin selected from a group consisting of cereulide, hemolysin BL (Hbl), non-hemolytic enterotoxin (Nhe) and cytotoxin K (CytK). The strain has wide application potential and can be applied to medicine and health care, feed additives or biological veterinary drugs as a probiotic candidate bacterial strain and used for removing pollutants in water bodies and preparing and purifying industrial enzymes.

Description

technical field [0001] The invention belongs to the field of microecological preparations, and in particular relates to a strain of non-toxic Bacillus cereus and its application. Background technique [0002] Probiotics have a wide range of potential applications in many fields such as medicine and health care, agriculture, food processing, feed industry and environmental protection, and occupy an important share in the booming biological industry. Safe, efficient and stable bacterial strains are the core content of the industrialization process of the whole probiotics. [0003] Bacillus cereus is an aerobic facultative anaerobic Gram-positive bacillus with oval-shaped spores and high temperature resistance. It can be used to treat diarrhea. The main mechanism is: 1) After the live bacterial preparation enters the intestinal tract, it forms a dominant flora, competes with harmful bacteria for nutrients and attachment sites, and competitively inhibits the growth of harmful b...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/742A61P31/04A23K1/16A01N63/00A01P21/00A01P1/00C12R1/085
Inventor 朱奎
Owner 朱奎
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