The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of action
By: Date: 2021-03-10 Categories: foodtechnology Tags: ,
  The result is”A rapid method and mechanism to identify the active compounds in Malus micromalus Makino fruit with spectrum-effect relationship, components knock-out and molecular docking technology” was published in”Food and Chemical Toxicology” magazine (immediate IF:5.759, JCR TOP Q2 area)
The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of actionimage
   Malus micromalus Makino, Malus micromalus Makino, of the Rosaceae, has a rich variety of fruit chemical components, including flavonoids, organic acids and volatile components. Pharmacological research It shows that flavonoids have anti-tumor, prevent cardiovascular and cerebrovascular aging, lighten liver spots, anti-inflammatory, anti-oxidant and anti-aging effects.
The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of actionimage(1)< br/>
  Figure 1 Xifu Begonia Fruit span>

   In this study, the effect of the fruit of Begonia xifuensis on tyrosinase activity was used to construct a”spectrum-effect model” combined with HPLC fingerprints, and screened by the spectrum-effect relationship Active substances, and use component knock-out method and UPLC-MS/MS method to identify the structure of the target component.
The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of actionimage(2)< br/>
  Fig.2 HPLC fingerprints of M. micromalus fruit and its reference fingerprint (R)
The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of actionimage(3)< br/>
  Fig.3 Standardization regression coefficient of PLSA model of characteristics common peak-tyrosinase inhibition activity. div>

The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of actionimage(4)< br/>
  Fig.4 The high resolution mass spectra of Peak 2 knocked-out component.
The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of actionimage(5)< br/>
  Fig.5 The high resolution mass spectra of Peak 8 knocked-out component.
The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of actionimage(6)< br/>
  Fig.6 The MS/MS fragmentation pathway of p-Coumaric acid (a), div>


  4-O-β-Glucopyranosyl-cis-coumaric acid (c).

   On this basis, the enzymatic reaction kinetics is used to explore the influence of knock-out components on the tyrosinase catalyzed reaction and the kinetic model analysis, and with the help of molecular docking The technology predicts the binding mode of active ingredients and tyrosinase, and clarifies the active ingredients that inhibit/activate tyrosinase in the fruit of Begonia xifuensis and their mechanism of action.
The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of actionimage(7)< br/>
  Fig. 7 Lineweaver-Burk plots of tyrosinase after the addition of p-coumaric acid (a), phloridzin (b ),quercetin-3-O-α-rhamnoside (c) and 4-O-β-glucopyranosyl-cis-coumaric acid (d).
The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of actionimage(8)< br/>

  Fig. 8 Molecular docking interaction of tyrosinase with p-coumaric acid(a), phloridzin(b),quercetin-3-O-α-rhamnoside(c) and 4-O-β-glucopyranosyl-cis-coumaric acid(d)
The National Edible Fungus Processing Technology Research and Development Center has made progress in the research of spectrum-effect relationship, component knockout combined with molecular docking technology to quickly identify active ingredients and the mechanism of actionimage(9)< br/>
  Note:(a, b, c, d) Plane projection diagram of the interaction between compound and tyrosinase.

   Through the study of spectrum-effect relationship, it is found that the fruit of Begonia xifuensis has the best inhibitory effect on tyrosinase activity at a concentration of 0.5 g/mL ; PLS-DA analysis showed that the inhibitory effects of P2, P3, P5 and P6 on tyrosinase activity in the characteristic chromatogram of Xifu crabapple fruit were positively correlated (R>0.1); the inhibitory effects of P8 and P9 on tyrosinase activity showed a positive correlation. Negative correlation (R<-0.1). P2, P6, P8, P10, P11, and 12 were identified by UPLC-MS/MS as p-coumarin acid, ferulic acid-acyl glucoside, coumarin acid-4-O-glucoside, and phallokin-2, respectively '-Xylose glucoside, phlorizin and quercetin-3-O-rhamnoside. The results of tyrosinase kinetics experiments showed that coumaric acid (P2) and phlorizin (P11) (P11>0.50 mmol/L) have a competitive inhibitory effect on tyrosinase, and the concentration of phlorizin is less than 0.25 When mmol/L, it has a mixed inhibitory effect. Coumaric acid-4-O-glucoside (P8) and quercetin-3-O-rhamnoside (P12) are mainly expressed in the concentration range of 0-9 mmol/L and 0.3-11 mmol/L, respectively Non-competitive activation type, while quercetin-3-O-rhamnoside (P12) is a mixed activation type. The molecular docking results of tyrosinase showed:p-coumaric acid (P2), coumaric acid-4-O-glucoside (P8), phlorizin (P11), quercetin-3-O-rhamnoside ( P12) is located in the active center of the hydrophobic pocket of the enzyme, which binds to tyrosinase residues by hydrogen bonds, and has a hydrophobic interaction with numerous surrounding hydrophobic residues to maintain the structure of the complex together.

  Using spectrum-effect relationship, ingredient knockout and high-resolution mass spectrometry identification technology to quickly identify active ingredients in complex traditional Chinese medicine systems, expound the overall view of traditional Chinese medicine, and explore the fields of active ingredients/ingredient groups , The team has formed a mature research strategy. The research results have been published in Food and Chemical Toxicology (https://doi.org/10.1016/j.fct.2019.110754);

  Food Science and Human Wellness (https://doi.org/10.1016/j.fshw.2021.02.019);

  Food and Chemical Toxicology (https://doi.org/10.1016/j.fct.2019.110754);

  International Journal of Molecular Sciences (https://doi.org/10.3390/ijms19113439);

  Frontiers in pharmacology (https://doi.org/10.3389/fphar.2020.01342) and other magazines. And with this technological innovation and integration, it won the second prize of Henan Science and Technology Progress Award in 2020.