Streptococcus mutans inhibitor and preparation method

A Streptococcus mutans and inhibitor technology, applied in the field of Streptococcus mutans inhibitors and its preparation, can solve problems affecting the cost of antibacterial drug preparation and use safety

Inactive Publication Date: 2018-11-02
BRIGHT DAIRY & FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Secondly, from the perspective of culture medium, most of the reports use chemical synthesis medium as fermentation base material for biological metabolism or direct synthesis, which will directly affect the preparation cost and safety of antibacterial drugs

Method used

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  • Streptococcus mutans inhibitor and preparation method
  • Streptococcus mutans inhibitor and preparation method
  • Streptococcus mutans inhibitor and preparation method

Examples

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preparation example Construction

[0025] When faced with the technical problem of "how to provide a new antimicrobial agent for Streptococcus mutans", the inventor, through creative work, provides a method for preparing an inhibitor of Streptococcus mutans in a specific embodiment, comprising the following steps (a) inoculating Paenibacillus bovis CGMCC No.8333 in the bran medium to ferment to obtain a fermented liquid; (b) centrifuging the fermented liquid obtained in step (a) to get the supernatant, placing Inactivate the protease in a high-temperature water bath. After cooling, slowly add ammonium sulfate to the supernatant, centrifuge to take the precipitate, add water to dissolve, and freeze-dry to obtain the crude product 1; (c) separate the crude product 1 obtained in step (b) through a gel column , collect, detect, and combine antibacterial active components, and freeze-dry to obtain crude product 2; (d) purify crude product 2 obtained in step (c) by HPLC, collect, detect, and combine antibacterial acti...

Embodiment 1

[0044] 1. Materials and methods

[0045] (a) Preparation of seeds (fermentation strains): dissolve the lyophilized powder of Paenibacillus bovis CGMCC No.8333 (the source of this bacterial strain is referring to the Chinese patent whose publication number is CN 103740618A) with a small amount of sterile distilled water, and inoculate One ring was drawn and streaked on TYC solid medium (purchased from OXOID Co., UK), cultured aerobically at 30°C for 48 hours, taken out, and a single colony was picked with an inoculation loop and put into 10 mL of TYC liquid medium (purchased from OXOID Co., U.K.), use a vortex shaker to evenly disperse the colony in the liquid medium, take it out after 24 hours of shaking culture at 30°C and 180rpm, and inoculate it into the TYC liquid medium (purchased from OXOID Co., UK), 30°C, 180rpm shaking culture for 24 hours, the culture was centrifuged at 15,000rpm for 10 minutes, the supernatant was discarded, the bacteria were washed twice with steril...

Embodiment 2

[0060] 1. Materials and methods

[0061] (a) Preparation of seed (fermentation strain): same as Example 1.

[0062] (b) Preparation of bran medium: fully mix 6% bran with distilled water, sterilize at 95° C. for 20 minutes, and cool to room temperature to obtain the bran medium with the required concentration.

[0063] (c) Detection methods of gel column separation, HPLC purification and antibacterial activity of samples: the same as in Example 1.

[0064] 2. Preparation of Streptococcus mutans inhibitors

[0065] The seeds of Paenibacillus bovis CGMCC No.8333 were aseptically inoculated in 6% (w / w) bran medium with an inoculum size of 1% (v / v), cultivated at 35°C and 100rpm for 96 hours to obtain a fermentation broth.

[0066] Centrifuge the fermentation broth at 10,000g for 10min to take the supernatant, put it in a water bath at 80°C for 10min to inactivate the protease, after cooling, slowly add ammonium sulfate to the supernatant to make the saturation of ammonium sulfa...

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Abstract

The invention belongs to the technical field of biology and specifically discloses a preparation method of a streptococcus mutans inhibitor. The preparation method comprises the following steps: (a) inoculating paenibacillus bovis CGMCC No.8333 into a bran culture medium to be fermented to obtain fermentation liquor; (b) centrifuging the fermentation liquor obtained in the step (a), taking supernatant, putting the supernatant into high-temperature water bath to inactivate protease, adding ammonium sulfate into supernatant after cooling, centrifuging to take precipitate, adding water to dissolve the precipitate and freeze drying to obtain a crude product 1; (c) performing gel column separation on the crude product 1 obtained in the step (b), collecting antibacterial active ingredients and freeze drying to obtain a crude product 2; (d) performing HPLC purification on the crude product 2 obtained in the step (c), collecting antibacterial active ingredients and freeze drying to obtain thestreptococcus mutans inhibitor. By means of the preparation method disclosed by the technical scheme of the invention, varieties and sources of the streptococcus mutans inhibitor are widened; the prepared inhibitor can effectively inhibit streptococcus mutans, staphylococcus aureus and the like and has a better application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an inhibitor of Streptococcus mutans and a preparation method thereof. Background technique [0002] Dental caries, commonly known as caries, tooth decay, is a bacterial disease, can be secondary to pulpitis and periodontitis, and even cause alveolar bone and jaw inflammation. There are many factors that cause dental caries, such as bacteria, oral environment (food, saliva), host, etc. Among them, bacteria are a necessary condition for the occurrence of dental caries. It is generally believed that there are two types of cariogenic bacteria, one is acid-producing bacteria, and the main one is Streptococcus mutans, Actinomyces and Lactobacillus can decompose carbohydrates to produce acid, leading to demineralization of inorganic matter in teeth; the other is Gram-positive cocci, which can destroy organic matter and cause dental caries to form after long-term action. Among ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P1/04A61P31/04A61P1/02C12R1/01
CPCA61K35/74C12P1/04C12R2001/01C12N1/205
Inventor 吴正钧冯华峰高彩霞韩瑨花榜清
Owner BRIGHT DAIRY & FOOD
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