A kind of preparation method of bacteriostatic Paenibacillus fermented liquid extract

A technology of bacillus and fermentation liquid, applied in the field of microorganisms, can solve problems such as teratogenicity, deficiency, food poisoning, etc., and achieve high-efficiency antibacterial effects, broad-spectrum antibacterial effects, and effects of inhibiting or killing pathogenic bacteria

Active Publication Date: 2020-05-19
BRIGHT DAIRY & FOOD CO LTD
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  • Abstract
  • Description
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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to overcome the chemically synthesized preservatives in the field of traditional Chinese medicine and food that will affect the original flavor of the food and also have potential carcinogenicity, teratogenicity or food poisoning to the human body. Hidden dangers, lack of natural and safe bacteriostatic agents, this technical problem provides a preparation method of bacteriostatic Paenibacillus fermentation liquid extract

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  • A kind of preparation method of bacteriostatic Paenibacillus fermented liquid extract
  • A kind of preparation method of bacteriostatic Paenibacillus fermented liquid extract
  • A kind of preparation method of bacteriostatic Paenibacillus fermented liquid extract

Examples

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Embodiment 1

[0044] Example 1 Extraction of antibacterial substances derived from Paenibacillus polymyxa (Paenibacillus polymyxa) BD3736

[0045] 1. Cultivation of BD3736 strain

[0046] The BD3736 strain preserved in 20% glycerol was inoculated on LB solid medium, and placed in an incubator at 30°C for 48 hours under atmospheric conditions. Then, a ring of single colony was scraped and placed in 15 mL of sterilized skim milk with a solid content of 10%, and incubated at a constant temperature of 30° C. and 180 r / min for 24 hours to obtain a BD3736 strain seed fermentation broth.

[0047] In a 500mL conical flask, 130mL of sterilized skim milk with a solids content of 10% was loaded, and the above-mentioned BD3736 strain seed fermentation liquid was inoculated according to the inoculation amount of 4% (v / v), and was shaken at a constant temperature of 30°C and 180r / min for 5 days. Obtain BD3736 strain fermentation broth.

[0048] 2. Preparation of BD3736 strain fermentation broth extract...

Embodiment 2

[0050] The antibacterial spectrum of embodiment 2 BD3736 bacterial strain fermented liquid extract

[0051] 1. Preparation of indicator bacteria plate

[0052] The pathogenic bacteria preserved in 20% glycerol were respectively transferred to BHI solid medium, and cultured at 30°C for 48h. Then a ring of single colony was scraped and transferred to liquid BHI, and cultivated at 30°C and 180r / min for 24h. Centrifuge at 12000rpm for 5min to collect the bacterial cells, dilute with sterile water to make bacterial suspension. Use a hemocytometer to count and control the final bacterial concentration to 10 6 cfu / mL.

[0053] After the BHI solid medium is cooled to 55°C, mix it with the prepared indicator bacteria suspension and pour it into a plate, and pour about 20mL of medium into each petri dish. After solidification and sufficient drying, the indicator bacteria plate can be obtained.

[0054] 2. Antibacterial spectrum of BD3736 strain fermentation broth extract

[0055] ...

Embodiment 3

[0057] Example 3 Purification of BD3736 strain fermentation broth extract

[0058] (1) The crude product of the fermentation liquid extract of the BD3736 strain described in Example 1 was dialyzed overnight at 4° C. using dialysis bags with cut-off values ​​of 3500 Da and 1000 Da respectively, and the samples in the bags were collected. Freeze-dry the dialysate to obtain freeze-dried powder.

[0059] (2) The lyophilized powder obtained in step (1) was dissolved in ultrapure water to a mass concentration of 200 mg / mL, and the sample volume was 10 mL, and separated by Sephadex G-15 (column volume about 800 mL) gel column chromatography. The eluent was ultrapure water with pH 7.0, and the flow rate was 3 mL / min. Collect one tube every 4min, about 12mL per tube. It was detected at 280nm with a protein UV detector. See the experimental results figure 2 .

[0060] The samples collected above were evaporated to dryness of the aqueous phase. Determination of the diameter of the...

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Abstract

The invention provides a preparation method of a bacteriostatic Paenibacillus sp. fermentation broth extract. The preparation method comprises the following steps: culturing Paenibacillus polymyxa CGMCC (China General Microbiological Culture Collection Center) No.10062 in cow milk; collecting supernatant of the fermentation broth to obtain the Paenibacillus sp. fermentation broth extract. The bacteriostatic Paenibacillus sp. fermentation broth extract prepared by the preparation method provided by the invention has a broad-spectrum and efficient bacteriostatic effect, and can be used for effectively restraining or killing pathogenic bacteria.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a preparation method of a bacteriostatic Paenibacillus fermentation liquid extract. Background technique [0002] During the process of processing, preservation, and transportation, food is easily contaminated by external microorganisms and deteriorates. Traditional methods of preserving food are usually pickling, smoking, cellar storage, and wine stains. With the continuous development of science and technology, chemically synthesized preservatives have become one of the most important additives in the food industry. However, studies have found that these chemically synthesized preservatives not only affect the original flavor of food, but also have potential safety hazards such as carcinogenicity, teratogenicity, or food poisoning to the human body. Therefore, finding and developing can effectively inhibit or kill food. Natural and safe antimicrobial agents ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P1/04C12R1/01
Inventor 高彩霞刘振民吴正钧崔燕丽吴江吴申懋
Owner BRIGHT DAIRY & FOOD CO LTD
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