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Agrobacterium ZX09, water-soluble beta-glucan prepared from Agrobacterium ZX09 and preparation method thereof and application on reducing blood sugar

A technology of Agrobacterium and strains, applied in the field of Agrobacterium sp.ZX09, which can solve the problems of increased production cost, increased difficulty of β-glucan, and restrictions on large-scale use, and achieves low cost, simple extraction method, and unique properties. Effect

Active Publication Date: 2012-05-23
SICHUAN HETAI SYNLIGHT BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But all these solutions increase the difficulty of obtaining the required β-glucan, greatly increase the cost of preparation, and limit the large-scale use in the field of production

Method used

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  • Agrobacterium ZX09, water-soluble beta-glucan prepared from Agrobacterium ZX09 and preparation method thereof and application on reducing blood sugar
  • Agrobacterium ZX09, water-soluble beta-glucan prepared from Agrobacterium ZX09 and preparation method thereof and application on reducing blood sugar
  • Agrobacterium ZX09, water-soluble beta-glucan prepared from Agrobacterium ZX09 and preparation method thereof and application on reducing blood sugar

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Prepare culture medium, NaH 2 PO 4 0.5g, KNO 3 2g, CaCl 2 0.07g, MgCl 2 0.2g, FeSO 4 ·7H 2 O 0.005g, MnSO 4 0.001g, ZnCl 2 0.005g, sucrose 20g, H 2 O1000mL, pH 5.0. The culture solution was sterilized by heating at 121°C for 15 minutes, cooled to 30°C, inoculated with 200 μL of colony liquid on a plate, and incubated at 28°C for 24 hours to form a seed solution. Add 2% seed culture solution to the culture solution that has been sterilized (sterilized at 121° C. for 15 minutes) and cooled to 30° C., and fermented at 28° C. for 48 hours. Two times the volume of ethanol was added to the fermentation broth to obtain crude β-glucan.

[0054] Mice fasted for 15 hours were directly fed polysaccharide 200mg / kg -1 Or feed polysaccharide mixed with starch 200mg / kg -1 , can effectively control the rise of blood sugar after meals.

Embodiment 2

[0056] Prepare culture medium, NaH 2 PO 4 0.5g, KNO 3 2g, CaCl 2 0.07g, MgCl 2 0.2g, FeSO 4 ·7H 2 O 0.0075g, MnSO 4 0.002g, ZnCl 2 0.005g, NaCl 20g, sucrose 20g, H 2 O1000mL, pH 7.2. The culture solution was sterilized by heating at 121°C for 15 minutes, cooled to 30°C, inoculated with 100 μL of colony liquid on a plate, and incubated at 28°C for 24 hours to form a seed solution. Add 10% seed culture solution to the culture solution that has been sterilized (sterilized at 121° C. for 15 minutes) and cooled to 30° C., and fermented at 28° C. for 72 hours. Add twice the volume of 95% ethanol to the fermentation broth to obtain a precipitate which is crude polysaccharide, take out the precipitate, redissolve it in water, and use the sevag method to remove protein three times. Add 2 volumes of ethanol to the deproteinized supernatant to precipitate polysaccharides, and redissolve the precipitate in water. The obtained polysaccharide aqueous solution is separated b...

Embodiment 3

[0058] Prepare culture medium, NaH 2 PO 4 1.0g, KNO 3 2.5g, CaCl 2 0.07g, MgCl 2 0.2g, FeSO 4 ·7H 2 O 0.0075g, MnSO 4 0.002g, ZnCl 2 0.005g, NaCl 20g, sucrose 30g, H 2 O1000mL, pH 7.2, heat the culture solution at 121°C to sterilize for 15 minutes, cool to 30°C, insert 200μL of plate colony liquid, and incubate at 30°C for 24h to form a seed solution. Add 4% seed culture solution to the culture solution that has been sterilized (sterilized at 121° C. for 15 minutes) and cooled to 30° C., and fermented at 28° C. for 48 hours. Add twice the volume of 95% ethanol to the fermentation broth to obtain a precipitate, which is the crude product, take out the precipitate, redissolve it in water, and use the sevag method to remove protein three times. Add 2 volumes of ethanol to the deproteinized supernatant to precipitate polysaccharides, and redissolve the precipitate in water. The obtained polysaccharide aqueous solution is separated by DEAE-32 cellulose and Sepharose...

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Abstract

The invention discloses Agrobacterium ZX09, water-soluble beta-glucan prepared from the Agrobacterium ZX09 and the preparation method thereof and the application on reducing blood sugar, wherein the bacterial strain of the Agrobacterium ZX09 is obtained by separating saline soil from Dongying city in Shandong province; the bacterial strain is Agrobacterium sp. and is collected in China Center for Type Culture Collection (CCTCC) on January 21st, 2010; and the collection number is CCTCC NO:M2010020. The structure general formula of the water-soluble beta-glucan prepared from the Agrobacterium ZX09 is as follows: ->(3)-beta-D-Glup-(1->3)-[beta-D-Glup-(1->3)-beta-D-Glup-]3-alpha-D-Glup-(1->3)-alpha-D-Glup-(1)n->, and n in the structural formula is from 100 to 200. The bacterial strain has the advantages of novelty, special properties, salt resistant growth, no pigment secretion in the process of growth, simple and convenient cultivation operation, simple technology, total synthetic media, definite components and low cost, and is suitable for industrial production.

Description

technical field [0001] The invention belongs to the technical field of microbes and microbial extracellular polysaccharides, in particular to an Agrobacterium sp. ZX09, a novel water-soluble β-glucan molecule produced by the bacterium, a preparation method thereof and an application in lowering blood sugar. Background technique [0002] β-glucan is widely distributed in fungi, bacteria and plants. At present, the β-glucan of medicinal and edible fungi mainly comes from the cell walls of edible and medicinal basidiomycete fungi and ascomycete yeasts. β-glucan in grains Sugars are mainly found in the cell walls of oat and barley endosperm, and the extraction of β-glucan from these substances is more complicated and the yield is lower. [0003] β-glucan not only exerts a variety of biological activities in various organisms, but also has multiple functions in the process of interaction between various organisms, and is an efficient biological response regulator (BRM). Accordin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C08B37/02C12P19/08A61K31/716A61P3/10C12R1/01
Inventor 张建法修爱慧朱斌
Owner SICHUAN HETAI SYNLIGHT BIOTECH LTD
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