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Peach gum polysaccharide degradation product PGP-1 and preparation method and application thereof

A technology of PGP-1 and peach gum polysaccharides, applied in biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., can solve difficult problems such as polysaccharide depolymerization, and achieve simple preparation, growth inhibition, and strong scavenging ability Effect

Active Publication Date: 2016-07-13
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the steric hindrance of the polysaccharide conformation, it is difficult for a single enzyme to completely depolymerize the polysaccharide

Method used

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  • Peach gum polysaccharide degradation product PGP-1 and preparation method and application thereof
  • Peach gum polysaccharide degradation product PGP-1 and preparation method and application thereof
  • Peach gum polysaccharide degradation product PGP-1 and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: Microbacterium degrades peach gum to prepare fermented liquid:

[0041] 1. Activation of strains

[0042] Store the strains on the slant at 4°C, streak the nutrient agar plate, incubate upside down at 37°C for 24 hours, pick a single clone from the plate and transfer it to the LB test tube culture medium, 6ml / branch, 200rpm, 37°C, and shake for 12 hours.

[0043] 1.8% LB liquid medium: 3.6g compound LB, dilute to 200ml with deionized water, PH=4, 121℃, sterilize for 20 minutes and set aside.

[0044] The strain is Microbacterium A5, which was preserved in the China Center for Type Culture Collection on August 7, 2009, and its preservation number is CCTCCNO:M209174;

[0045] 2. Shake flask culture

[0046] Inoculate the activated seed liquid into the peach gum shaker medium, 500ml Erlenmeyer flask, the liquid volume is 200ml, the initial peach gum concentration is 8%, the initial pH=4, the inoculum volume is 3%, and the culture temperature is 30°C , 160r...

Embodiment 2

[0048] Embodiment 2: the fermented liquid gained in embodiment 1 is separated and purified:

[0049] First, the fermented liquid prepared in Example 1 is mixed with Sevage reagent (chloroform: amyl alcohol or n-butanol (mixed in a 4:1 or 5:1 ratio)) in a certain ratio, and then centrifuged to remove the interface between the extract and the Sevage reagent. The denatured protein at place, and then adopt 75% ethanol to precipitate above-mentioned substance, obtain peach gum crude polysaccharide.

[0050] Choose DEAE-Cellulose52 anion exchange to further purify peach gum crude polysaccharide, as follows:

[0051]Add 10g of DEAE-52 to 500ml of water and soak for 24 hours to fully expand the cellulose particles and remove suspended fine particles. Then transfer to Buchner funnel (with 200-mesh nylon mesh inside) for suction filtration, soak in 200ml0.5mol / L NaOH–NaCl solution for 4h, transfer to Buchner funnel (with 200-mesh nylon mesh inside) for suction filtration, and distilled...

Embodiment 3

[0060] Example 3: Identification of PGP-1:

[0061] 1. Purity identification and molecular weight determination of PGP1

[0062] Using ShodexKS-802 and KS-804 series column, differential refraction detector, through Agilent1100series high performance liquid chromatography, with different relative molecular mass of dextran (P-5, P-10, P-20, P-50, P-100, P-200, P-400 and P-800) were used as standard items to make a standard curve to measure the purity and relative molecular mass of the polysaccharide.

[0063] Such as figure 2 As shown, there is a single peak with higher purity at 13.191min, indicating that PGP-1 is a homogeneous component with higher purity. Such as image 3 As shown, the relative molecular mass of PGP-1 is in the range of 1×10 5 ~1×10 6 In the range.

[0064] 2. IR spectrum analysis of PGP1

[0065] Weigh 5mg of PGP1 for KBr compression, at 400-4000cm -1 between scans.

[0066] Such as Figure 4 As shown, PGP-1 contains polysaccharides at 4000~400cm...

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Abstract

The invention provides a peach gum polysaccharide degradation product PGP-1.The peach gum polysaccharide degradation product PGP-1, with molecular weight between 1X10<5> and 1X10<6>, contains no neutral polysaccharide and consists of arabinose, xylose, mannose, glucose and galactose in a molar ratio of 0.637:0.054:0.073:0.022:0.556.The peach gum polysaccharide degradation product PGP-1 is simple to prepare and capable of promoting bifidobacterium growth remarkably, inhibiting enterococcus faecium growth remarkably, scavenging hydroxyl free radicals and DPPH free radicals effectively and inhibiting Hela cell growth remarkably; Galectin-3-mediated cell agglutination inhibiting experiments show that the peach gum polysaccharide degradation product PGP-1 is capable of remarkably inhibiting agglutination of chicken blood red blood cells, Hela cells and MCF-7 cells mediated by Galectin-3.

Description

technical field [0001] The invention belongs to the technical field of peach gum polysaccharide preparation, and more specifically relates to a peach gum polysaccharide degradation product PGP-1 and its preparation method and application. Background technique [0002] Peach gum is a resin secreted from the bark of the Rosaceae plant peach or mountain peach. The medicinal value of peach gum is widely recorded in ancient books of traditional Chinese medicine. Peach gum is flat in nature, sweet and bitter in taste, good at promoting blood circulation and reducing swelling, dredging stranguria and relieving pain, quenching thirst and quenching thirst. Clinical application has good effects on bloody stranguria, stone stranguria, diabetes and other diseases. [0003] my country is rich in peach gum resources, which is a high-quality traditional Chinese medicine resource. There are reports on the composition of peach gum polysaccharides at home and abroad, but the enzymatic hydr...

Claims

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Application Information

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IPC IPC(8): C08B37/00C12P19/14C12P19/04A61P1/00A61P35/00A61P39/06C12R1/01
CPCC08B37/0003C12P19/04C12P19/14
Inventor 冉艳红刘天祥李弘剑周天鸿王伟杨晓萍
Owner JINAN UNIVERSITY
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