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. 2019 Nov 7;9(1):16172.
doi: 10.1038/s41598-019-52711-7.

The integration of metabolome and proteome reveals bioactive polyphenols and hispidin in ARTP mutagenized Phellinus baumii

Henan Zhang  1 , Ruibing Chen  2 , Jingsong Zhang  1 , Qitao Bu  2 , Wenhan Wang  1 , Yanfang Liu  1 , Qing Li  3 , Ying Guo  2 , Lei Zhang  4   5 , Yan Yang  6
Affiliations

The integration of metabolome and proteome reveals bioactive polyphenols and hispidin in ARTP mutagenized Phellinus baumii

Henan Zhang et al. Sci Rep. .

Abstract

Phellinus baumii, also called "Sang Huang" in China, is broadly used as a kind of health food or folk medicine in Asia for its high biological activities, e.g. anti-tumor, anti-oxidation and anti-inflammatory activities. Although some previous studies have indicated that polysaccharides and flavonoids showed the activity of inhibiting tumor cells, the active metabolites of P. baumii needs further research. In our study, a stable P. baumii mutant (A67), generated by ARTP mutagenesis strategy, showed more significantly inhibiting tumor cells and enhancing antioxidant activity. Our further studies found that the increase of polyphenols content, especially hispidin, was an important reason of the biological activity enhancement of A67. According to the results of the integrated metabolome and proteome study, the increase of polyphenol content was caused by upregulation of the phenylpropanoid biosynthesis. This study expanded the understanding of active compounds and metabolic pathway of P. baumii.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Discovery of the A67 mutant strain with high polyphenols and flavonoids content. (A) Preparation of P. baumii protoplasts. (a and b), One hour after enzyme treatment; (c and d), three hours after enzyme treatment. (B) Comparison of polyphenols, flavonoids, glucose content and biomass between SH1 and A67. (C) Comparison of appearance characteristics of SH1 and A67. (D) The antagonistic reaction between A67 and SH1.
Figure 2
Figure 2
Biological activities of P. baumii ethanol extracts at different concentrations in vitro. (A) Inhibitory effects on tumor cell line HepG2. (B) Inhibitory effects on tumor cell line K562. Each value is expressed as means ± SD (n = 3). 5-fluorouracil (5-fu) served as a positive control and DMSO as negative control. (C) Comparison of antioxidant effects of P. baumii ethanol extracts at different concentrations by DPPH method. (D) Comparison of antioxidant effects of P. baumii ethanol extracts at different concentrations by FRAP and TEAC method.
Figure 3
Figure 3
Differences of metabolite accumulation between SH1 and A67 strains. (A) PCA scores plots of nine SH1 and nine A67 strains based on the extract spectral data of UPLC-LTQ Orbitrap and announced differential metabolites. (B) PLS-DA scores plots of nine SH1 and nine A67 strains based on the extract spectral data of UPLC-LTQ Orbitrap and announced differential metabolites. (C)The structure of 5 differential polyphenol metabolites. (D) The changes and relative content of five differential metabolites. Different colors represent different metabolites, the size of the circle represents the relative content and the Y-axis represents the fold changes.
Figure 4
Figure 4
Biological activities of hispidin. Inhibitory effects of hispidin at different concentrations on (A) HepG2 and (B) K562 cell lines in vitro. Antioxidant effects of hispidin at different concentrations by (C) DPPH method and (D) FRAP and TEAC method.
Figure 5
Figure 5
Differences of proteome between SH1 and A67 strains. (A) PLS-DA scores plots. (B) Number of differentially expressed proteins. (C) GO analysis of differential proteins. (D) Correlation network between differential metabolites and proteins.
Figure 6
Figure 6
The research strategy that integrates omics data to explore bioactive products and biosynthesis pathway analysis of hispidin. The solid lines represent the confirmed metabolic pathway. Dotted lines represent presumed metabolic pathway. The red arrows represent the upregulated steps in A67 strain. Dotted arrows represent the presumed steps. The histograms represent the relative expression level of the proteins in the SH1 and A67 strains.
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