Screening for proteins related to the biosynthesis of hispidin and its derivatives in Phellinus igniarius using iTRAQ proteomic analysis

Jinjing Guo,Xiuli Wu,Li Zhou,Xiaoxi Liu,Hongyan Ji,Yuanjie Li,Cheng Liu,Zhibo Jiang,Yu Huang,Changcai Bai

doi:10.1186/s12866-021-02134-0

Abstract

BackgroundHispidin (HIP) and its derivatives, a class of natural fungal metabolites, possess complex chemical structures with extensive pharmacological activities. Phellinus igniarius, the most common source of HIP, can be used as both medicine and food. However, the biosynthetic pathway of HIP in P. igniarius remains unclear and we have a limited understanding of the regulatory mechanisms related to HIP. In this work, we sought to illustrate a biosynthesis system for hispidin and its derivatives at the protein level.ResultsWe found that tricetolatone (TL) is a key biosynthetic precursor in the biosynthetic pathway of hispidin and that its addition led to increased production of hispidin and various hispidin derivatives. Based on the changes in the concentrations of precursors and intermediates, key timepoints in the biosynthetic process were identified. We used isobaric tags for relative and absolute quantification (iTRAQ) to study dynamic changes of related proteins in vitro. The 270 differentially expressed proteins were determined by GO enrichment analysis to be primarily related to energy metabolism, oxidative phosphorylation, and environmental stress responses after TL supplementation. The differentially expressed proteins were related to ATP synthase, NAD binding protein, oxidoreductase, and other elements associated with electron transfer and dehydrogenation reactions during the biosynthesis of hispidin and its derivatives. Multiple reaction monitoring (MRM) technology was used to selectively verify the iTRAQ results, leading us to screen 11 proteins that were predicted to be related to the biosynthesis pathways.ConclutionThese findings help to clarify the molecular mechanism of biosynthesis of hispidin and its derivatives and may serve as a foundation for future strategies to identify new hispidin derivatives.

Highlights

Hispidin (HIP) and its derivatives, a class of natural fungal metabolites, possess complex chemical structures with extensive pharmacological activities
To determine that the peaks at 19.7 min and 23.4 min were HIP and phelligridin D, the chemical structures of the two purified compounds A and B were assessed by Nuclear Magnetic Resonance (NMR) spectra and High resolution electrospray ionization mass spectroscopy (HRESIMS)
Compared with the peak position of standards in High performance liquid chromatography (HPLC) and the data from HRESIMS, the compound was speculated to be a combination of transhispidin and cis-hispidin, which was confirmed by 2D NMR

Summary

Introduction

Hispidin (HIP) and its derivatives, a class of natural fungal metabolites, possess complex chemical structures with extensive pharmacological activities. Edible and medicinal fungi are a common source of nutrients and medicines in Asian nations and are well recognized as an important source of biologically active compounds. Phellinus igniarius (DC.Ex Fr.) Quel, a wild macrofungi, contains many bioactive compounds with properties such as antibacterial, antioxidative, antitumor, and antimutagenic activities, and has been widely used in China, Japan, and Korea for many years [1,2,3]. Phelligridins are a class of chemical compounds from P. igniarius that possess radicalscavenging activity and a broad range of pharmacological properties, and includes chemicals such as Phelligridin D and Phelligridin G [6, 7]. Hispidin (6-(3,4-dihydroxystyryl)-4-hydroxy-2-pyrone, HIP), is an important polyphenol found in P. igniarius. HIP has attracted significant attention in the fields of chemistry, pharmacy, and microbiology [18,19,20,21]

Methods

Results

Discussion

Conclusion