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Cryptococcus Neoformans capsular polysaccharide preparation method

A technology of cryptococcus neoformans and capsular polysaccharides, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of high cost, difficulty in large-scale production, and small processing capacity, and achieve simplification of the preparation process Effect

Inactive Publication Date: 2013-03-06
天津贻诺琦生物工程有限公司
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantage of this method is that the operation is complicated, the processing volume is small, the cost is high, and it is not easy to scale production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1, using the preparation method of the present invention to extract the capsular polysaccharide of Cryptococcus neoformans, the steps are as follows:

[0023] Prepare 0.6L of YM broth medium, its composition is 1.8g yeast extract, 3g peptone, 1.8g maltose extract, 6g D-glucose. The culture temperature is 30° C., the shaking speed is 200 rpm, and the culture time is 36 h. The bacteria solution was sterilized at 121°C for 25 minutes, and centrifuged at 16,000g for 30 minutes to remove bacteria. Slowly add calcium acetate powder to the supernatant while stirring to a final concentration of 5%. Add glacial acetic acid to adjust the pH to about 5.0. Add three volumes of 95% ethanol to the above solution, a precipitate will appear immediately, and stand overnight at 4°C. Pour off most of the supernatant, and transfer the remaining liquid to a 50ml round bottom centrifuge tube. Centrifuge at 12,000g for 5 minutes. The supernatant was discarded, and the precipit...

Embodiment 2

[0024] Example 2, using the preparation method of the present invention, using cetyltrimethylammonium bromide to purify the crude capsular polysaccharide, the specific steps are as follows:

[0025] 1) Add NaCl powder to the crude capsular polysaccharide solution to a final concentration of 0.2M, slowly add CTAB powder while stirring at a rate of 3 times the total mass of the polysaccharide, let stand for 1 hour, and centrifuge at 23°C and 12,000g for 5 minutes , add about 15ml of 10% ethanol to resuspend the precipitate, centrifuge at 12,000g for 30 minutes at 23°C, discard the supernatant; repeat the ethanol wash twice; Absolute ethanol, a precipitate will be formed immediately, let it stand for 1 hour, 23°C, 12,000g centrifuge for 10 minutes, resuspend the precipitate with 70% ethanol, 23°C, 12,000g centrifuge for 10 minutes, discard the supernatant, the precipitate is the purified new type Cryptococcal capsular polysaccharide.

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PUM

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Abstract

The invention relates to the fungal polysaccharide preparation field, and concretely relates to a preparation method of Cryptococcus Neoformans capsular polysaccharide. The invention provides a method for preparing the Cryptococcus Neoformans capsular polysaccharide through utilizing cetyl trimethyl ammonium bromide. The method comprises the following steps: 1, processing a secretion supernatant containing the capsular polysaccharide through using calcium acetate and glacial acetic acid, and precipitating through using ethanol to prepare a crude capsular polysaccharide; and 2, processing a solution of the crude capsular polysaccharide through using the cetyl trimethyl ammonium bromide to obtain the purified capsular polysaccharide. The method has the advantages of simplicity, high efficiency, low cost, and effective avoiding of the use of toxic and harmful chemical reagents.

Description

technical field [0001] The invention relates to the field of polysaccharide preparation of fungi. In particular, the present invention relates to the preparation of capsular polysaccharides of Cryptococcus neoformans. Background technique [0002] Cryptococcus neoformans is an important opportunistic pathogenic fungus mainly found in aviary farms or soil contaminated with bird excrement, in dust and air in the rooms of AIDS and HIV-associated cryptococcosis patients The pathogen of Cryptococcus neoformans can also be found. Cryptococcus neoformans often infects immunocompromised or immunocompromised patients, causing clinical symptoms and even life-threatening. [0003] In recent years, fungal meningitis has increased significantly due to the long-term and extensive application of broad-spectrum antibacterial drugs, adrenal cortex hormones, and immunosuppressants after tumor organ transplantation, as well as the prevalence of AIDS. Cryptococcus neoformans meningitis (cryp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/02C12P19/04C12R1/01
Inventor 史东东何永胜周泽奇李宁粟艳王东东彭洁
Owner 天津贻诺琦生物工程有限公司
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