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Preparation method for konjac mannatide

A technology of konjac mannan and glycopeptides, which is applied in the field of preparation of glycopeptides and konjac mannan peptides, can solve problems such as unseen mannan peptides, and achieve the effect of broadening sources and broadening preparation methods

Inactive Publication Date: 2016-04-06
SHAANXI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no method for preparing mannan peptides from konjac powder

Method used

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  • Preparation method for konjac mannatide
  • Preparation method for konjac mannatide
  • Preparation method for konjac mannatide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 1) Fermentation: Add 1 g of konjac fly powder to 100 mL of fermentation medium, and use Bacillus pyrolyticum and Bacillus pumilus for symbiotic fermentation. After the fermentation is completed, the fermentation liquid is centrifuged at 8000r / min for 30 minutes at 4°C to remove the bacteria, the supernatant is added with 2 times of purified water, and the pH is adjusted to 3.8 with trichloroacetic acid, and the precipitation is removed after standing. The supernatant is the konjac polypeptide liquid.

[0032] Among them, the composition of the medium is: peptone 1g, beef extract 1.5g, NaNO 3 0.2g, K 2 HPO 4 0.2g, KCl0.1g, MgSO 4 ·7H 2 O0.1g, FeSO 4 0.005g, distilled water 100mL. The pH value of the medium is 7, the strain ratio of Bacillus pyrolyticum and Bacillus pumilus is 1:1, and the concentration of the bacterial solution is diluted to 5×10 7 Individual / mL, the inoculum size is 12% (V / V, the inoculum size refers to the volume fraction of the bacterial soluti...

Embodiment 2

[0037] 1) Fermentation: Add 3 g of konjac powder into 100 mL of fermentation medium, and use Bacillus pyrolyticum and Bacillus pumilus for symbiotic fermentation. After the fermentation is completed, the fermentation liquid is centrifuged at 9000r / min for 15 minutes at 4°C to remove the bacteria, the supernatant is added with 3 times of purified water, and the pH is adjusted to 3.9 with trichloroacetic acid, and the precipitation is removed after standing. The supernatant is the konjac polypeptide liquid.

[0038] Among them, the composition of the medium is: peptone 1g, beef extract 1.5g, NaNO 3 0.08g, K 2 HPO 4 0.25g, KCl0.03g, MgSO 4 ·7H 2 O0.08g, FeSO 4 0.01g, distilled water 100mL. The pH value of the medium is 8, the strain ratio of Bacillus pyrolyticum and Bacillus pumilus is 1:2, and the concentration of the bacteria solution is diluted to 8×10 7 individual / mL, the inoculum size is 10% (V / V), the fermentation temperature is 32° C., and the fermentation time is 4...

Embodiment 3

[0043] 1) Fermentation: Add 4 g of konjac powder into 100 mL of fermentation medium, and use Bacillus pyrolyticum and Bacillus pumilus for symbiotic fermentation. After the fermentation is completed, the fermentation broth is centrifuged at 10000r / min for 30min at 4°C to remove the bacteria, the supernatant is added with 4 times of purified water, the pH is adjusted to 4 with trichloroacetic acid, and the precipitation is removed after standing. The supernatant is the konjac polypeptide liquid.

[0044] Among them, the composition of the medium is: peptone 1g, beef extract 1.5g, NaNO 3 0.39g, K 2 HPO 4 0.04g, KCl0.05g, MgSO 4 ·7H 2 O0.05g, FeSO 4 0.001g, distilled water 100mL. The pH value of the medium is 8.5, the strain ratio of Bacillus pyrolyticum and Bacillus pumilus is 1:3, and the concentration of the bacteria solution is diluted to 7×10 8 individual / mL, the inoculum size is 8% (V / V), the fermentation temperature is 34°C, and the fermentation time is 44h.

[004...

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Abstract

The invention provides a preparation method for konjac mannatide. Flying konjac powder is used as the raw material and fermented with pyrolysis-resistant protein bacillus and bacillus pumilus to prepare konjac polypeptide; an ammonium sulfate fractional precipitation method is adopted to extract soluble glycopeptides in the konjac polypeptide; after dialysis and desalination, raw glycopeptides are subjected to DEAE-cellulose column chromatography and gel column chromatography in sequence for separation and purification; lastly, an affinity chromatography method is adopted to obtain purified konjac mannatide. The preparation method fills up the technical blank that the konjac mannatide is prepared from the flying konjac powder serving as the raw material in combination with a fermentation method, the ammonium sulfate fractional precipitation technology and the chromatography technology, opens up a new way for development and utilization of the flying konjac powder and production of products with high added value, and has great significance in the fields of food, medicine and the like.

Description

technical field [0001] The invention belongs to the field of microbial fermentation and medicinal peptides, and relates to a method for preparing glycopeptides, in particular to a method for preparing konjac mannan peptides. Background technique [0002] Mannan peptide used to be called polyanti-A, is the first biological immune enhancer in my country. It is a mixture formed by the combination of mannan peptide molecules with different chain lengths. There are no small molecular sugars and free amino acids. Its main structure is that the polypeptide chain is connected with mannan. Modern pharmacological and clinical studies have shown that mannan peptide has obvious biological activity, which can regulate the immune disorder of the body and significantly enhance autoimmunity. Now it is mostly used in the treatment of immunocompromised or immune disorders, such as recurrent colds, recurrent respiratory infections, leukopenia and adjuvant treatment of tumors to reduce the adv...

Claims

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Application Information

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IPC IPC(8): C12P39/00C12P21/00C07K1/36C07K1/30C07K1/22C07K1/16C12R1/01C12R1/07
Inventor 毛跟年贾莹张诗韵许牡丹
Owner SHAANXI UNIV OF SCI & TECH
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