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Preparation method of bacteroides fragilis capsular polysaccharide A

A technology of Bacteroides fragilis and capsular polysaccharides, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of toxic and harmful organic solvents, unfavorable production safety and environmental protection, etc., to promote shedding, gain The effect of increasing the rate and reducing the toxicity

Pending Publication Date: 2021-08-10
GUANGZHOU ZHIYI PHARMA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] But in essence, the above traditional methods rely on phenol, which is toxic and harmful, and its use is not conducive to production safety and environmental protection

Method used

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  • Preparation method of bacteroides fragilis capsular polysaccharide A
  • Preparation method of bacteroides fragilis capsular polysaccharide A
  • Preparation method of bacteroides fragilis capsular polysaccharide A

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0037] The invention provides a method for preparing Bacteroides fragilis capsular polysaccharide A, the preparation method comprising the following steps:

[0038] Prepare a bacterial suspension of Bacteroides fragilis with pH ≤ 5, extract by heating, and collect the extract to obtain a crude sugar solution of capsular polysaccharide A.

[0039]In the preparation method provided by the present invention, the specific preparation method of the bacterial suspension of pH≤5 Bacteroides fragilis is not particularly limited, including but not limited to the following methods: (1) cultivating Bacteroides fragilis and collecting the collected thalli (can be Called bacteria slime, water content can be controlled at 50% ~ 95%) resuspended in a solvent such as water (for example, resuspended in purified water whose quality is 3 to 10 times the weight of the thallus), and then added to adjust the pH The pH of the bacterial suspension was adjusted to ≤5. The pH regulator can be any one ...

Embodiment 1

[0053] Embodiment 1, the preparation of Bacteroides fragilis bacterium slime

[0054] Streak inoculation of Bacteroides fragilis ZY-312 strain on blood plate, anaerobic culture for 48h. Observe the colony morphological characteristics, staining characteristics, size, club shape and distribution, etc.

[0055] Colony characteristics: After Bacteroides fragilis ZY-312 was cultured on a blood plate at 37°C for 48 hours, it was slightly convex, translucent, white, smooth, non-hemolytic, and the diameter of the colony was between 1mm and 3mm. See figure 1 .

[0056] Morphology under the microscope: Bacteroides fragilis ZY-312 was examined by Gram staining. It is a Gram-negative bacterium with a typical rod shape, blunt rounded ends and dense staining. The uncolored part in the middle of the bacteria is like a vacuole. figure 2 .

[0057] Select a single colony and inoculate it in tryptone broth for fermentation and culture for 8 hours (at a temperature of 37°C). The obtained ba...

Embodiment 2

[0058] Example 2, comparison of preparation methods of Bacteroides fragilis capsular polysaccharide A

[0059] The bacteria slime prepared in Example 1 was used to carry out the experiment.

[0060] (1) Method 1: Prepare Bacteroides fragilis capsular polysaccharide A using the method provided by the invention

[0061] ①Take 50g of bacteria slime (obtained by Example 1), add 300g of purified water to resuspend the bacteria, adjust its pH to 3.5 with 1mol / L hydrochloric acid solution, extract at 100°C for 1.5h, cool to room temperature, and centrifuge at 12000g for 10min at room temperature. Take the supernatant to obtain a crude sugar solution.

[0062] ②The crude sugar solution is concentrated by 10KD ultrafiltration membrane ultrafiltration to remove small molecular impurities until the conductivity is stable, and the reflux liquid is collected;

[0063] ③Add an equal volume of 40mmol / L Tris-HCl (pH8.5) to the reflux liquid to convert salt; DEAE Sepharose FastFlow ion excha...

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Abstract

The invention relates to a preparation method of bacteroides fragilis capsular polysaccharide A. The preparation method comprises the following steps: preparing a bacterial suspension of bacteroides fragilis with the pH value of less than or equal to 5, heating and extracting, and collecting an extracting solution to obtain a crude sugar solution of capsular polysaccharide A. Compared with the prior art, the preparation method has the following beneficial effects that the inventor accidentally finds that the capsular polysaccharide A can be effectively promoted to fall off from the bacteroides fragilis thalli by heating and extracting the bacteroides fragilis bacterial suspension of which the pH value is less than or equal to 5, so that the capsular polysaccharide A is extracted. Compared with a traditional phenol / water method, the capsular polysaccharide A can be obtained under the condition that phenol is not used, and the toxicity is low. Moreover, by adopting the preparation method, the yield of the bacteroides fragilis capsular polysaccharide A is obviously improved.

Description

technical field [0001] The invention relates to the preparation and application technology of Bacteroides fragilis capsular polysaccharide, in particular to a preparation method of Bacteroides fragilis capsular polysaccharide A. Background technique [0002] Bacteroides fragilis (bacteroides fragilis) is a member of the genus Bacteroides among Gram-negative anaerobic bacteria, belonging to the phylum Bacteroidetes, which is completely different from bifidobacteria and lactic acid bacteria of the phylum Firmicutes. There are 25 species of Bacteroides, 10 species from humans only, 10 species from animals only, and 5 species from both humans and animals. Bacteroides fragilis is an obligate anaerobic bacterium, which can be divided into enterotoxigenic Bacteroides fragilis (ETBF)) and non-enterotoxigenic Bacteroides fragilis (ETBF) according to whether it can synthesize and secrete Bacteroides fragilis enterotoxin (BFT) Toxin-type Bacteroides fragilis (Nontoxigenic Bacteroides ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C12N1/20C08B37/00C12R1/01
CPCC12P19/04C12N1/20C08B37/0003
Inventor 邝高波王从峰易晓敏黄海清李平王晔
Owner GUANGZHOU ZHIYI PHARMA INC
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