Genetic evidence for the requirements of antroquinonol biosynthesis byAntrodia camphorataduring liquid-state fermentation

Abstract

The solid-state fermentation of Antrodia camphorata could produce a variety of ubiquinone compounds, such as antroquinonol (AQ), AQB, 4-acetyl-AQB and so on. However, ubiquinone compounds are hardly synthesized during liquid-state fermentation (LSF) and the mechanism of synthesis remain unclear. To investigates the influence of different precursors on the synthesis of AQ, three exogenous precursors (ubiquinone 0, UQ0; farnesol and farnesyl diphosphate, FPP) were used in LSF. The results showed that UQ0 successfully induced AQ production; however, farnesol and FPP did not play a role in producing AQ. The results of fermentation experiments show that the precursor that restricts the synthesis of AQ is the quinone ring, not the isoprene side chain. An efficient Agrobacterium-mediated transformation system of A. camphorata was established. Exogenous genes were effectively integrated into the genome of A. camphorata using this system. Then, we overexpressed a series of genes for quinone ring modification (coq2-6) and isoprene synthesis (HMGR, fps) by using the ATMT transformation system, which endogenously increased the content of intracellular precursor for AQ biosynthesis in A. camphorata. The results showed that overexpression of genes for isoprene side chain synthesis could not increase the yield of AQ, but overexpression of coq2 and coq5 could significantly increase AQ production. This is consistent with the results of the fermentation experiment with the addition of precursors. It indicated that the A. camphorata lack the ability to modify the quinone ring of AQ during LSF. Of the modification steps, prenylation of UQ0 is the key step of AQ biosynthesis during LSF. The result will help us to understand the genetic evidence for the requirements of antroquinonol biosynthesis in A. camphorata.

A review on the cultivation, bioactive compounds, health-promoting factors and clinical trials of medicinal mushrooms Taiwanofungus camphoratus, Inonotus obliquus and Tropicoporus linteus

“…Furthermore, bioassay-guided isolation from methanolic extracts of T. camphoratus revealed a potent anti-inflammatory compound, 4-acetylantroquinonol B ( 42 ), which significantly inhibited polyinosinic-polycytidylic acid-induced NO production by RAW264.7 macrophages [ 72 ]. Hence, due to the highly potent anti-tumor properties and demonstrated bioactivities of antroquinonol, many studies have been conducted to enhance its synthesis [ 73 – 78 ].…”

Isolation and identification of bioactive compounds from Antrodia camphorata against ESKAPE pathogens

“…Additionally, the addition of petroleum ether extracts from camphor trees to the culture medium has been shown to significantly increase the total triterpene yield of A. camphorata. Therefore, deep fermentation technology stands out due to its advantages, including short cycles, high efficiency, low cost, and ease of scalable production [43][44][45]. In this study, A. camphorata was cultured using liquid fermentation because it is less time-consuming and cost-effective.…”

Review of Bioactivity, Isolation, and Identification of Active Compounds from Antrodia cinnamomea

“…However, almost no antroquinonol is obtained under liquid-state fermentation conditions [ 57 ]. When ubiquinone 0 is used as a precursor in liquid-state fermentation, antroquinonol production can be successfully induced [ 58 ]. Given the low content of antroquinonol in A. cinnamomea , the preparative isolation of antroquinonol remains a difficult process.…”