Hiascic acid

Natural products of lichen-forming fungi are structurally diverse and have a variety of medicinal properties. Despite this, they have limited implementation in industry mostly because the corresponding genes are unknown for most of their natural products. Here, we implement a long-read sequencing and bioinformatic approach to identify the putative biosynthetic gene cluster of the bioactive natural product gyrophoric acid (GA). Using 15 high-quality genomes representing nine GA-producing species of the lichen-forming fungal genus Umbilicaria, we identify the most likely GA cluster and investigate the cluster gene organization and composition across the nine species. Our results show that GA clusters are promiscuous within Umbilicaria, and only three genes are conserved across species, including the polyketide synthase (PKS) gene. In addition, our results suggest that the same cluster codes for different, but structurally similar compounds, namely, GA, umbilicaric-, and hiascic acid, bringing new evidence that lichen metabolite diversity is also generated through regulatory mechanisms at the molecular level. Ours is the first study to identify the most likely GA cluster and, thus, provides essential information to open new avenues for biotechnological approaches to producing and modifying GA and similar lichen-derived compounds. GA PKS is the first tridepside PKS to be identified. IMPORTANCE The implementation of natural products in the pharmaceutical industry relies on the possibility of modifying the natural product (NP) pathway to optimize yields and pharmacological effects. Characterization of genes and pathways underlying natural product biosynthesis is a major bottleneck for exploiting the medicinal properties of the natural products. Genome mining is a promising and relatively cost- and time-effective approach to utilize unexplored NP resources for drug discovery. In this study, we identify the most likely gene cluster for the lichen-forming fungal depside gyrophoric acid in nine Umbilicaria species. This compound shows cytotoxic and antiproliferative properties against several cancer cell lines and is also a broad-spectrum antimicrobial agent. This information paves the way for generating GA analogs with modified properties by selective activation/deactivation of genes.

In the present study, we characterized the phytochemical properties, which were specifically associated with phenolic compounds and antioxidant activities in six distinct ecotypes of Umbilicaria aprina Nyl. from Iran (including Kivarestan, Mishan, Takht-e Nader, Tochal, Sabalan, and Sahand) to detect diversities within the species. Total phenolic concentration (TPC) and radical scavenging capacities of U. aprina ecotypes were evaluated. Moreover, qualitative differences between chemical profiles were surveyed using liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). Relatively moderate TPCs (Kivarestan = 36.12 ± 2.1, Mishan = 41.59 ± 2.2, Takht-e Nader = 31.85 ± 1.3, Tochal = 37.55 ± 2.3, Sabalan = 28.91 ± 2.5, and Sahand = 31.59 ± 2.2) were observed for ecotypes, but a very strong correlation (r = -0/842) was obtained between TPCs and IC50 values. Based on the results of LC-ESI-MS/MS, the following chemical substances were identified: orsellinic acid (1), lecanoric acid (2), evernic acid (3), gyrophoric acid (4), umbilicaric acid (5), hiascic acid (6), stictic acid (7) methyl hiascic acid (8), and an unknown substance (9). The MS/MS fragmentation scheme for each substance was determined and proposed. Wide discrepancies were observed in the chemical profiles of lichen ecotypes, which may corroborate the influence of ecological locality conditions, for example, altitude and slope aspects on secondary metabolism of lichen species U. aprina. The north-facing and east-facing ecotypes (Sabalan and Mishan, respectively) lacked depsidones (stictic acid) mainly because they receive the least direct radiation. Mishan ecotype, as the only east-facing ecotype, showed the most different chemical profile.

Depsides and tridepsides are secondary metabolites found in lichens. In the last 10 years, there has been a growing interest in the pharmacological activity of these compounds. This review aims to discuss the research findings related to the biological effects and mechanisms of action of lichen depsides and tridepsides. The most studied compound is atranorin, followed by gyrophoric acid, diffractaic acid, and lecanoric acid. Antioxidant, cytotoxic, and antimicrobial activities are among the most investigated activities, mainly in in vitro studies, with occasional in silico and in vivo studies. Clinical trials have not been conducted using depsides and tridepsides. Therefore, future research should focus on conducting more in vivo work and clinical trials, as well as on evaluating the other activities. Moreover, despite the significant increase in research work on the pharmacology of depsides and tridepsides, there are many of these compounds which have yet to be investigated (e.g., hiascic acid, lassalic acid, ovoic acid, crustinic acid, and hypothamnolic acid).