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

Abstract

Background Hispidin (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. Results We 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. Conclution These 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.

Hispidin‐enriched Sanghuangporus sanghuang mycelia SS‐MN4 ameliorate disuse atrophy while improving muscle endurance

“…The extracted sample was sterilized through 0.22‐μm syringe for in vitro and in vivo studies. For hispidin purification, the extraction protocol was conducted according to previous publication with slight modification 20 . In brief, the extract was partitioned using hexane–90% MeOH (2:1) mixture and centrifuged at 3600 × g for 5 min.…”

“…For hispidin purification, the extraction protocol was conducted according to previous publication with slight modification. 20 In brief, the extract was partitioned using hexane-90% MeOH (2:1) mixture and centrifuged at 3600 × g for 5 min. This procedure is repeated two times, and the hispidin was concentrated using the rotary evaporator.…”

Pilot Study: Nutritional and Preclinical Safety Investigation of Fermented Hispidin-Enriched Sanghuangporus sanghuang Mycelia: A Promising Functional Food Material to Improve Sleep

“…Hispidin was isolated according to a previous study with slight modification ( 26 ). Solvents such as methanol, formic acid, acetonitrile, and n-hexane used for extraction and chromatographic separation were analytical grade (Merck, Darmstadt, Germany).…”

Narrative Review: Bioactive Potential of Various Mushrooms as the Treasure of Versatile Therapeutic Natural Product

“…Ganoderma tsugae Polysaccharides [256,257] Gomphus clavatus Ergosterol, phenolic compounds [258,259] Grifola frondosa Phenolic compounds, β-1,6 and β-1,3-glucan [260] Helvella crispa Phenolic compounds [261] Hericium erinaceus Phenolic compounds [262] Hydnum repandum Tocopherols, phenolic compounds [263,264] I. obliquus p-Hydroxybenzoic acid, quercetin, kaempferol [265,266] Laccaria laccata Tocopherols, phenolic compounds [267] Lactarius citriolens Free sugars, fatty acids, tocopherols, and phenolic acids [268] Lactarius deliciosus Phenolic compounds, flavonoids [269][270][271] Lactarius piperatus Phenolic compounds, flavonoids [236,272] Lactarius salmonicolor Phenolic compounds [273,274] Lentinula edodes Gallic acid, protocatechuic acid, catechin, tocopherols [275][276][277] Lepista nuda β-Carotene, α-tocopherol [278][279][280][281] Leucopaxillus giganteus β-carotene, ascorbic acid, lycopene, phenolic compounds [282] Macrolepiota procera Phenolic compounds [283] Marasmius oreades Flavonoids, ascorbic acid [284,285] Meripilus giganteus Gallic, p-coumaric, protocatechin, caffeic, and vanillc acids [286][287][288] Phellinus igniarius Hispidin [289,290] Phellinus linteus β-Tocopherol, protocatechuic acid, gallic acid; pyrogallol; homogentisic acid, αand β-glucans [291] Pleurotus ostreatus β-Glucans galli...…”