Application of lentinan in preparing drug for inhibiting activity of alpha-glucosaccharase
A technology of glucosidase and lentinan, which is applied in the directions of drug combination, organic active ingredients, pharmaceutical formulations, etc., can solve the problem that α-glucosidase active drugs have not been reported, and achieve the effect of preventing diabetes and reducing postprandial blood sugar.
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Embodiment 1
[0021] Example 1 The inhibitory effect of lentinan on the activity of α-glucosidase
[0022] Take 40 μL α-glucosidase (final concentration is 4 μM) and 40 μL different concentrations of lentinan solution (final concentration is 0.3, 0.6, 0.9, 1.2, 1.5, 1.8, 2.1, 2.4, 2.7 mM), at 37 ° C, pH = Incubate in the phosphate buffer solution of 6.8 for 10 minutes, then add 40 μL p-nitrophenyl-α-D-glucopyranoside (final concentration is 8 mM), put it in a constant temperature shaker at 37°C for 60 minutes, and add 120 μL 0.2mol·L -1 Sodium carbonate solution, its absorbance was measured at 405nm. At the same time, the inhibitor test group, the inhibitor control group, the enzyme activity test group and the enzyme activity control group were made, as shown in Table 1. Each experiment was repeated more than 3 times. Calculate α-glucosidase inhibitory percentage=[1-(A by the following formula 抑制剂实验组 -A 抑制剂对照组 ) / (A 酶活实验组 -A 酶活对照组 )] × 100%. Depend on figure 1 It can be seen that wi...
Embodiment 2
[0026] Example 2 Judgment of the inhibition type of lentinan to α-glucosidase
[0027] Take 40 μL of different concentrations of lentinan (final concentrations: 0, 0.9, 1.5, and 2.1 mM), add 40 μL of α-glucosidase (final concentrations: 1, 2, and 4 μM) respectively, and put them in a constant temperature shaker at 37°C for 10 minutes. Then add 40 μL of p-nitrophenyl-α-D-glucopyranoside (final concentration: 8 mM), react at 37°C for 60 min, and add 120 μL after the reaction to a concentration of 0.2 mol L -1 Sodium carbonate solution, its absorbance was measured at 405nm. Linear fitting was carried out with different enzyme concentrations and reaction speeds. As the concentration of different lentinan increased, the slope of the line increased and all lines passed the origin, and the type of reversible inhibition could be judged. Depend on figure 2 It can be seen that the straight line a is the blank group (the final concentration of lentinan is 0); the straight lines b-d ar...
Embodiment 3
[0028] Embodiment 3 Lentinan is to the Lineweaver-Burk curve of α-glucosidase inhibitory effect
[0029] Take 40 μL of different concentrations of lentinan (final concentration: 0, 0.9, 1.5, 2.1 mM), add 40 μL α-glucosidase (final concentration: 4 μM) respectively, put them in a constant temperature shaker at 37°C for 10 minutes, and then put them in 40 μL p-nitrophenyl-α-D-glucopyranoside (final concentration: 2, 4, 8 mM), react at 37°C for 60 min, after the reaction, add 120 μL with a concentration of 0.2mol L -1 Sodium carbonate solution, its absorbance was measured at 405nm. Using Lineweaver-Burk's double reciprocal plot, if you get a set of straight lines with different slopes and intersecting on the Y axis, it is competitive inhibition; if you get a set of straight lines with different slopes and intersecting on the X axis, it is non-competitive inhibition ; If you get a set of straight lines with different slopes that intersect in the second or third quadrant, it is mi...
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