ALKANNIN AND ITS INHIBITORY ACTIVITY AGAINST STARCH HYDROLASES
Keywords:
Alkannin, Ultrasonic-assisted extraction, Single-factor experiment, Response surface methodology, α-Amylase, α-Glucosidase, Molecular dockingAbstract
This study optimized the ethanol-based ultrasonic-assisted extraction process of alkannin and evaluated its inhibitory activity and mechanism against α-amylase and α-glucosidase. The suitable ranges for various factors were determined through single-factor experiments. Subsequently, the Plackett-Burman design was employed to screen for significant factors, combined with the steepest ascent design to approach the optimal region, and the Box-Behnken design was utilized to optimize the extraction conditions. Finally, enzyme inhibition assays and molecular docking were conducted to investigate its activity and mechanism. The results indicated that the liquid-to-solid ratio, ultrasonic time, and ethanol volume fraction were significant influencing factors. The optimal extraction conditions were determined as follows: a liquid-to-solid ratio of 10:1 (mL/g), an ethanol volume fraction of 90.5%, an ultrasonic time of 45 min, and 2 extraction cycles. Under these conditions, the extraction yield of alkannin reached 1.952%. The half-maximal inhibitory concentration (IC₅₀) values of alkannin against α-amylase and α-glucosidase were 2.729 μM and 1.409 μM, respectively. Molecular docking revealed that alkannin exhibited non-competitive inhibition against α-amylase (binding energy: -6.9 kcal·mol⁻¹) and competitive inhibition against α-glucosidase (binding energy: -7.8 kcal·mol⁻¹). This study provides an experimental basis for the extraction process optimization and enzymatic inhibitory activity of alkannin.References
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