Fermented crude extract of streptococcus salivarius, and preparation method and application thereof

A technology of streptococcus salivarius and crude extract, applied in the field of fermented crude extract of streptococcus salivarius and its preparation, can solve problems such as adverse reactions, imbalance of oral flora, limited research, etc., and achieves widening sources, good stability, Significant effect of inhibitory activity

Active Publication Date: 2017-12-08
BRIGHT DAIRY & FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, most of the research reports on dental caries treatment focus on controlling or reducing the number of Streptococcus mutans in the oral cavity. However, a healthy oral environment is generally in a state where harmful bacteria and beneficial bacteria reach a balance. Excessive reduction of harmful bacteria will Bring the imbalance of oral flora, and then cause other adverse reactions of the human body
In addition, some clinically used bactericidal drugs against Streptococcus mutans, such as silver ammoniu

Method used

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  • Fermented crude extract of streptococcus salivarius, and preparation method and application thereof
  • Fermented crude extract of streptococcus salivarius, and preparation method and application thereof
  • Fermented crude extract of streptococcus salivarius, and preparation method and application thereof

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preparation example Construction

[0021] In a specific embodiment, the present invention provides a method for preparing a fermented crude extract of Streptococcus salivarius, the method comprising the following steps: (1) inoculating Streptococcus salivarius CGMCC NO.13308 in sucrose-containing M17 culture Ferment in base to obtain fermented liquid, centrifuge the obtained fermented liquid to get the supernatant; (2) add ammonium sulfate to the supernatant obtained in step (1), and centrifuge to collect ammonium sulfate when the saturation is 40-80%. The precipitate was dissolved, dialyzed, and freeze-dried.

[0022] The above-mentioned technical scheme adopts Streptococcus salivarius (Streptococcus salivarius) CGMCCNO.13308 for the first time, takes M17 medium as the culture medium, and centrifuges, precipitates with ammonium sulfate, dialyzes, and freeze-dries to prepare Streptococcus mutans biofilm formation inhibitory effect. The crude extract discloses that the fermented crude extract of Streptococcus sa...

Embodiment 1

[0033] 1. Materials and methods

[0034] (1) Preparation of seeds (fermentation strains):

[0035] Streptococcus salivarius CGMCC No.13308: Dissolve the freeze-dried powder of Streptococcus salivarius CGMCC No.13308 with a small amount of sterile distilled water, and get a ring with an inoculation loop and streak it on M17 solid medium (purchased from OXOID Co., UK), After anaerobic culture at 37°C for 24 hours, single colonies were picked with an inoculation loop and put into 10 mL of M17 liquid medium (purchased from OXOID Co., UK). Take out after 24 hours of oxygen culture, inoculate in M17 liquid medium (purchased from OXOID Co., UK) with 2% (v / v) inoculum, after 24 hours of anaerobic culture at 37 ° C, the culture is centrifuged at 15000 rpm for 10 minutes, and the supernatant is discarded. After washing the bacterium twice with sterile distilled water, suspend it with the original culture volume of sterile distilled water to obtain seeds for fermentation. The bacterial ...

Embodiment 2

[0043] 1, material and method: with embodiment 1

[0044] 2. Preparation and activity detection of crude extract of Streptococcus mutans biofilm inhibitor

[0045] Streptococcus salivarius CGMCC No.13308 seeds were aseptically inoculated in the M17 liquid medium containing sucrose 5% (w / v) with an inoculum size of 1% (v / v), and cultured anaerobically at 42°C for 24h to obtain a fermentation broth. The obtained fermentation broth was centrifuged at 12000 g for 10 min to obtain the supernatant.

[0046] Slowly add ammonium sulfate to the above supernatant, centrifuge at 12000g for 10min to collect the precipitate precipitated when the saturation of ammonium sulfate is 40-60%. After the dialysis is completed, the solution in the dialysis bag is freeze-dried to obtain the crude extract B of the Streptococcus mutans biofilm inhibitor.

[0047] After determination, the inhibition rate of 10 mg / mL crude extract B solution to Streptococcus mutans biofilm formation was 58%.

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Abstract

The invention provides a fermented crude extract of streptococcus salivarius, and a preparation method and application thereof. The preparation method comprises the following steps of (1) inoculating streptococcus salivarius CGMCC NO.13308 into an M17 culture medium containing saccharose, fermenting to obtain fermentation liquor, and centrifuging the obtained fermentation liquor to obtain a supernate; (2) adding ammonium sulfate into the supernate obtained through the step (1), centrifugally collecting sediments precipitated when the saturability of the ammonium sulfate is 40 to 80 percent, dissolving the obtained sediments, carrying out dialysis, freezing and drying to obtain the fermented crude extract of the streptococcus salivarius. The invention adopts a streptococcus salivarius CGMCC NO.13308 strain for preparing the fermented crude extract for the first time, the fermented crude extract of the streptococcus salivarius has an inhibitory effect on a streptococcus mutans biological membrane, and discloses that the streptococcus salivarius CGMCC NO.13308 has a new function of being fermented to produce a streptococcus mutansbiological membrane forming inhibitor, and the source of the streptococcus mutans biological membrane forming inhibitor is widened.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a fermented crude extract of Streptococcus salivarius and its preparation method and application. 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. If it is not treated in time, the lesion will continue to develop, forming cavities, and eventually the crown will be completely destroyed and disappeared. The final result of its development is tooth loss. Dental caries is characterized by high incidence and wide distribution. The disease is the main common disease of the oral cavity and one of the most common human diseases. The World Health Organization has listed it as one of the three major diseases of human beings, along with tumors and cardiovascular diseases. [0003] There are many factors that cause dental cari...

Claims

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Application Information

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IPC IPC(8): A61K35/744A61P31/04C12P1/04C12R1/46
CPCA61K35/744C12P1/04C12N1/205C12R2001/46
Inventor 韩瑨吴正钧冯华峰
Owner BRIGHT DAIRY & FOOD
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