Peach gum polysaccharide degradation product PGP-2 as well as preparation method and application thereof

A PGP-2, peach gum polysaccharide technology, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve problems such as difficult polysaccharide depolymerization, and achieve simple preparation, wide application prospects, and growth inhibition. Effect

Active Publication Date: 2016-06-15
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-2 as well as preparation method and application thereof
  • Peach gum polysaccharide degradation product PGP-2 as well as preparation method and application thereof
  • Peach gum polysaccharide degradation product PGP-2 as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: Microbacterium degrades peach gum to prepare fermented liquid:

[0040] 1. Activation of strains

[0041] 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 12h.

[0042] 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.

[0043] 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;

[0044] 2. Shake flask culture

[0045] 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 , 160rpm sh...

Embodiment 2

[0047] Embodiment 2: the fermented liquid obtained in embodiment 1 is separated and purified:

[0048] 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.

[0049] Choose DEAE-Cellulose52 anion exchange to further purify peach gum crude polysaccharide, as follows:

[0050]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 distill...

Embodiment 3

[0059] Example 3: Identification and analysis of PGP-2

[0060] 1. IR spectrum analysis of PGP-2

[0061] Weigh 5mg of PGP2 for KBr tableting, at 400-4000cm -1 between scans.

[0062] Such as figure 2 As shown, PGP-2 is at 3430cm -1 O─H sugar ring stretching vibration absorption peak is very small, 2888cm -1 The place is C─H (mainly including CH, CH 2 、CH 3 ) stretching vibration absorption peak, 1500~1200cm -1 Several absorption peaks exist at (1243cm -1 、1359cm -1 、1468cm -1 ) is C-H variable angle vibration, 1110cm -1 There is a variable angle vibration of C-O-C at 961cm -1 The peak at may be caused by the vibration of D-glucose, 843cm -1 The position is the C-H variable angle vibration of the pyranose β type.

[0063] 2. MALDI-TOF-MS analysis of PGP-2:

[0064] Take 1 mg of PGP-2 polysaccharide sample and dissolve it in 10 μl of TFA for MALDI-TOF-MS analysis, the results are as follows image 3 shown.

[0065] Depend on image 3 It is concluded that the f...

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Abstract

The invention provides a peach gum polysaccharide degradation product PGP-2. The PGP-2 is obtained by obtaining a fermentation broth by inoculating a peach gum culture medium with microbacterium A5 and performing separation and purification; the microbacterium A5 is preserved, with the preservation number of CCTCC NO:M209174 in the CCTCC (China Center for Type Culture Collection) on August 7, 2009; anion exchange is adopted for separation and purification, and a NaCl solution is adopted for gradient elution. The PGP-2 is simple to prepare, can significantly promote growth of Bifidobacterium while inhibiting growth of Enterococcus faecium, has higher scavenging activity for hydroxyl free radicals and DPPH free radicals and can significantly inhibit growth and proliferation of Hela cells, Galectin-3 mediated cell agglutination experiments indicate that the PGP-2 can significantly inhibit agglutination of Galectin-3 mediated red blood cells, Hela cells and MCF-7 cells.

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-2 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 冉艳红王伟李弘剑周天鸿杨晓萍王雅秋
Owner JINAN UNIVERSITY
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