Alpha-1,4-glucan and its preparation method and use
A technology of glucan and crude polysaccharide, which is applied in the directions of pharmaceutical formulations, medical preparations containing active ingredients, organic active ingredients, etc.
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Embodiment 1
[0044] Example 1: Preparation of α-1,4-glucan LJW0F2
[0045] a. Polysaccharide extraction:
[0046] Dried honeysuckle (produced in Pingyi County, Shandong Province, purchased from Shanghai Yanghetang Pharmaceutical Chain Operation Co., Ltd.), degreased with 95% ethanol for one week, and then dried naturally at room temperature. The dried honeysuckle 1000g was extracted 5 times with 20L of boiling water, 6h each time. Sulfuric acid-phenol detects no obvious reaction, filter, combine the extracts each time, heat and concentrate to 3L, add about 500g trichloroacetic acid after cooling to make the final concentration 15%, deproteinize at 4°C, and then centrifuge , the supernatant was neutralized by 1mol / L NaOH to a pH value of 7.0, and then dialyzed against flowing water for 72 hours, the liquid in the dialysis bag was concentrated to 2L volume, and three times the volume of 95% ethanol was added under stirring, and stood overnight, poured The supernatant was removed, centrifug...
experiment Embodiment 1
[0049] Experimental Example 1: Polysaccharide Structure Analysis:
[0050] Show that the relative weight-average molecular weight of LJW0F2 is about 37.1kDa through high performance gel chromatography (HPGPC) analysis, and its purity determination figure is shown in figure 1 . Sugar composition analysis showed that LJW0F2 was a dextran. Infrared spectrum shows, 3403cm -1 O-H stretching vibration absorption peak, 2927cm -1 C-H stretching vibration absorption peak, 1000-1400cm -1 Vibration signals of C-O and sugar ring nearby, 1720cm -1 There is no absorption peak nearby, indicating that the polysaccharide does not contain uronic acid ( figure 2 ). 13 In the C NMR spectrum, the carbon signal at δ100.98 is the C-1 signal of α-glucan. Other carbon signals are C-2 (δ72.97), C-3 (δ74.60), C-4 (δ78.08), C-5 (δ72.61) and C6 (δ61.70) ( image 3 ).
[0051] The types and proportions of sugar residue linkages in polysaccharide LJW0F2 can be analyzed by methylation. The results...
experiment Embodiment 2
[0056] Inhibit Aβ 42 aggregation test
[0057] 1) Thioflavin fluorescent labeling experiment
[0058] Aβ 42 The powder (Rpeptide Company) was dissolved in 110 μL of anhydrous DMSO (concentration: 2 mM) to make a mother solution. Dissolve 1 μL of this solution in 19 μL phosphate buffer (50 mM phosphate, pH 7.5, 100 mM NaCl, 0.02% NaN 3 ), or 1 μL of this solution was added with 10 μL of LJW0F2 polysaccharide solution of different concentrations (0.5 mg / mL, 1.0 mg / mL, 2.0 mg / mL) and then 9 μL of PBS buffer, incubated at 37 °C for 30 min, and then added with thioflavin solution (6.25 μM thioflavin dissolved in 50mM glycine-NaOH, pH 8.5) 80μL, incubated at 37°C, and detected with a microplate reader (Novostar, BMG labtech company) every 2h, the detection wavelength was Ex=450 / 10nm, Em=483 / 10nm.
[0059] Depend on Figure 4A It can be seen that α-1,4-glucan LJW0F2 can completely inhibit Aβ at a final concentration of 100 μg / mL 42 gathering.
[0060] 2) Atomic force microsc...
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