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
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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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