Concordance of multigene genealogy along with morphological evidence unveils five novel species and two new records of boletoid mushrooms (fungi) from India.

Agaricales, Russulales and Boletales are dominant orders among the wild mushrooms in Basidiomycota. Boletaceae, one of the major functional elements in terrestrial ecosystem and mostly represented by ectomycorrhizal symbionts of trees in Indian Himalaya and adjoining hills, are extraordinarily diverse and represented by numerous genera and species which are unexplored or poorly known. Therefore, their hidden diversity is yet to be revealed. Extensive macrofungal exploration by the authors to different parts of Himalaya and surroundings, followed by through morphological studies and multigene molecular phylogeny lead to the discovery of five new species of wild mushrooms: Leccinellum bothii sp. nov., Phylloporus himalayanus sp. nov., Phylloporus smithii sp. nov., Porphyrellus uttarakhandae sp. nov., and Retiboletus pseudoater sp. nov. Present communication deals with morphological details coupled with illustrations and phylogenetic inferences. Besides, Leccinellum sinoaurantiacum and Xerocomus rugosellus are also reported for the first time from this country.

Factors affecting composition of fatty acids in wild-growing forest mushrooms.

The importance of mushrooms as a food source is continually increasing. To investigate how environmental factors affect the nutritional value of mushrooms, we harvested them in eastern Poland, south-central Germany, and northwestern Belgium in plots with similar environmental conditions but varying in tree species composition and richness. We used gas chromatography-mass spectrometry (GC-MS) to analyze the fatty acid (FA) content of the mushrooms. Fungal species identity explained the largest part (40%) of the total variation in FA concentration and composition. Environmental factors accounted for 1-12% of variation. The concentration of FA, especially saturated fatty acids, decreased with increasing understory cover and increasing nitrogen concentration in the topsoil. The effect of tree species richness or tree species identity was negligible. Our results suggest that the nutritional value of mushrooms depends mainly on the species identity of fungi, but that their FA content is slightly higher in forests with less undergrowth and in nitrogen-poor soils.

A revised phylogeny of Boletaceae using whole genome sequences.

The porcini mushroom family Boletaceae is a diverse, widespread group of ectomycorrhizal (ECM) mushroom-forming fungi that so far has eluded intrafamilial phylogenetic resolution based on morphology and multilocus data sets. In this study, we present a genome-wide molecular data set of 1764 single-copy gene families from a global sampling of 418 Boletaceae specimens. The resulting phylogenetic analysis has strong statistical support for most branches of the tree, including the first statistically robust backbone. The enigmatic Phylloboletellus chloephorus from non-ECM Argentinian subtropical forests was recovered as a new subfamily sister to the core Boletaceae. Time-calibrated branch lengths estimate that the family first arose in the early to mid-Cretaceous and underwent a rapid radiation in the Eocene, possibly when the ECM nutritional mode arose with the emergence and diversification of ECM angiosperms. Biogeographic reconstructions reveal a complex history of vicariance and episodic long-distance dispersal correlated with historical geologic events, including Gondwanan origins and inferred vicariance associated with its disarticulation. Together, this study represents the most comprehensively sampled, data-rich molecular phylogeny of the Boletaceae to date, establishing a foundation for future robust inferences of biogeography in the group.

Amanitins: The Most Poisonous Molecules of the Fungal World.

Among the toxic metabolites of the fungal world, those that, due to their strong biological effect, can seriously (even fatally) damage the life processes of humans (and certain groups of animals) stand out. Amatoxin-containing mushrooms and the poisonings caused by them stand out from the higher fungi, the mushrooms. There are already historical data and records about such poisonings, but scientific research on the responsible molecules began in the middle of the last century. The goals of this review work are as follows: presentation of the cosmopolitan mushroom species that produce amanitins (which are known from certain genera of four mushroom families), an overview of the chemical structure and specific properties of amanitins, a summary of the analytical methods applicable to them, a presentation of the "medical history" of poisonings, and a summary of the therapeutic methods used so far. The main responsible molecules (the amanitins) are bicyclic octapeptides, whose structure is characterized by an outer loop and an inner loop (bridge). It follows from the unusual properties of amanitins, especially their extreme stability (against heat, the acidic pH of the medium, and their resistance to human, and animal, digestive enzymes), that they are absorbed almost without hindrance and quickly transported to our vital organs. Adding to the problems is that accidental consumption causes no noticeable symptoms for a few hours (or even 24-36 h) after consumption, but the toxins already damage the metabolism of the target organs and the synthesis of nucleic acid and proteins. The biochemical catastrophe of the cells causes irreversible structural changes, which lead to necrotic damage (in the liver and kidneys) and death. The scientific topicality of the review is due to the recent publication of new data on the probable antidote molecule (ICR: indocyanine green) against amanitins. Further research can provide a new foundation for the therapeutic treatment of poisonings, and the toxicological situation, which currently still poses a deadly threat, could even be tamed into a controllable problem. We also draw attention to the review conclusions, as well as the mycological and social tasks related to amanitin poisonings (prevention of poisonings).

Refinement and complete solution NMR analysis of the structure of a 6-branched 1,3-ß-D-glucan (OL-2) isolate from Omphalialapidescens.

OL-2 is a water-soluble ß-glucan produced by Omphalia lapidescens. This versatile glucan has potential applications in various industries, including food, cosmetics, and pharmaceuticals. In addition, OL-2 is known for its promising applications as a biomaterial and drug, owing to its reported antitumor and antiseptic properties. Although the biological activities of ß-glucans vary depending on their primary structure, holistic clarification of OL-2 via solution NMR spectroscopy to ascertain its complete and unambiguous structure has not yet been achieved. In this study, a chain of solution NMR techniques, such as correlation spectroscopy, total correlation spectroscopy (TOCSY), nuclear Overhauser effect and exchange spectroscopy, 13C-edited heteronuclear single quantum coherence (HSQC), HSQC-TOCSY, heteronuclear multiple bond correlation, and heteronuclear 2-bond correlation pulse sequences were used to unambiguously assign all 1H and 13C atoms in OL-2. Based on our investigation, OL-2 consists of a 1,3-ß-glucan backbone chain decorated with a single 6-branched ß-glucosyl side unit on every fourth residue.

Second world record for two Calvatia species (Agaricaceae: Basidiomycota) / Segundo registro mundial para duas espécies de Calvatia (Agaricaceae: Basidiomycota)

Calvatia is a genus of gasteroid fungi, comprising about 47 species worldwide. In this paper we report the second worldwide occurrence of two poorly known species of Calvatia, recorded in the Cerrado biome of Brazil: C. oblongispora and C. nodulata. Detailed morphological descriptions and illustrations, including scanning electron micrographs of hyphae and basidiospores are provided, as well a discussion on their taxonomy and geographic distribution.

Calvatia é um gênero de fungos gasteroides que compreende cerca de 47 espécies em todo o mundo. Neste artigo relatamos a segunda ocorrência de duas espécies pouco conhecidas de Calvatia, registradas no bioma Cerrado do Brasil: C. oblongispora e C. nodulata. Descrições morfológicas detalhadas e ilustrações são fornecidas, incluindo micrografias eletrônicas de varredura de hifas e basidiósporos, bem como uma discussão sobre sua taxonomia e distribuição geográfica.

A phylogenetic overview of Squamanita, with descriptions of nine new species and four new combinations.

Nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS barcode) sequence data from eight type specimens of previously described Squamanita species were obtained. Phylogenetic analysis of ITS and partial nuc 28S rDNA data revealed Squamanita as paraphyletic splitting into two monophyletic groups, which we recognize as the genera Squamanita and Dissoderma. We accept 14 Squamanita and nine Dissoderma species, provide the first sequences of 13 of these, and describe six new species of Squamanita and three new species of Dissoderma. We transfer three species of Squamanita into Dissoderma, one into Cystoderma, and treat S. basii and S. umbilicata as synonyms of D. paradoxum. Squamanita can be distinguished from Dissoderma by the generally larger fleshier basidiomata with a tricholomatoid or amanitoid stature and yellowish to tawny brown pileus and often similarly colored stipe. Most species have cheilo- and pleurocystidia. Species of Dissoderma are small, collybioid or mycenoid, lack cystidia, and the pileus and often upper stipe are purplish gray. Both genera parasitize basidiomata of other agarics.

Proteomic Research on the Antitumor Properties of Medicinal Mushrooms.

Medicinal mushrooms are increasingly being recognized as an important therapeutic modality in complementary oncology. Until now, more than 800 mushroom species have been known to possess significant pharmacological properties, of which antitumor and immunomodulatory properties have been the most researched. Besides a number of medicinal mushroom preparations being used as dietary supplements and nutraceuticals, several isolates from mushrooms have been used as official antitumor drugs in clinical settings for several decades. Various proteomic approaches allow for the identification of a large number of differentially regulated proteins serendipitously, thereby providing an important platform for a discovery of new potential therapeutic targets and approaches as well as biomarkers of malignant disease. This review is focused on the current state of proteomic research into antitumor mechanisms of some of the most researched medicinal mushroom species, including Phellinus linteus, Ganoderma lucidum, Auricularia auricula, Agrocybe aegerita, Grifola frondosa, and Lentinus edodes, as whole body extracts or various isolates, as well as of complex extract mixtures.

Treasures from the forest: Evaluation of mushroom extracts as anti-cancer agents.

Mushrooms provide a reliable source of bioactive compounds and have numerous nutritional values, which is one of the reasons why they are widely used for culinary purposes. They may also be a remedy for several medical conditions, including cancer diseases. Given the constantly increasing number of cancer incidents, the great anticancer potential of mushrooms has unsurprisingly become an object of interest to researchers. Therefore, this review aimed to collect and summarize all the available scientific data on the anti-cancer activity of mushroom extracts. Our research showed that mushroom extracts from 92 species, prepared using 12 different solvents, could reduce the viability of 38 various cancers. Additionally, we evaluated different experimental models: in vitro (cell model), in vivo (mice and rat model, case studies and randomized controlled trials), and in silico. Breast cancer proved to be sensitive to the highest number of mushroom extracts. The curative mechanisms of the studied mushrooms consisted in: inhibition of cancer cell proliferation, unregulated proportion of cells in cell cycle phases, induction of autophagy and phagocytosis, improved response of the immune system, and induction of apoptotic death of cells via upregulation of pro-apoptotic factors and downregulation of anti-apoptotic genes. The processes mainly involved the expression of caspases -3, -8, -9, AKT, p27, p53, BAX, and BCL2. The quoted results could lead to the classification of mushrooms as nutraceuticals used to prevent a variety of disorders or to support treatment of cancer diseases.

Molecular systematics and taxonomic overview of the bird's nest fungi (Nidulariaceae).

Fungi in the Nidulariaceae, otherwise known as 'bird's nest fungi', are among the least studied groups of Agaricomycetes (Basidiomycota). Bird's nest fungi are globally distributed and typically grow on woody debris or animal dung as saprotrophs. This group of fungi is morphologically diverse with ca. 200 described species. Phylogenetic relationships of bird's nest fungi were investigated with four commonly used loci (ITS, LSU, tef, and rpb2). The family was resolved as a monophyletic group with Squamanitaceae as a potential sister taxon. Cyathus and Crucibulum each formed its own independent and well-supported clade. Nidula and Nidularia formed a clade together, but each genus is polyphyletic. Two Mycocalia species included in our analyses were on their own separate branches, indicating that this genus is also polyphyletic. Misidentifications were detected in most genera, suggesting that species concepts need to be revisited and refined throughout Nidulariaceae. Several bird's nest fungi species have global geographical distributions whereas others may have more limited ranges. Basic morphological characters of bird's nest fungi have likely been lost or gained multiple times. The phylogenetic placement of Crucibulum is unclear and the sister lineage of bird's nest fungi is not conclusive. Further studies with data from rare species and additional informative genes are needed to fully resolve the topology of Nidulariaceae and identify its sister group with more certainty.

An untold story of a novel mushroom from tribal cuisine: an ethno-medicinal, taxonomic and pharmacological approach.

India showcases an array of fascinating and rare mushrooms that grow exclusively in the wilderness of West Bengal. Thus, the state has always been our prime choice to document myco-diversity and associated indigenous knowledge. Fortuitously, a recent expedition gifted us a violet-coloured Russuloid macrofungus, called "Jam Patra", that plays an integral part in the food security of local ethnic groups. However, the species has not received the much-needed attention among city dwellers and remains abandoned, motivating us to carry a thorough investigation. To our surprise, extensive analyses on morphological features and nrITS based phylogenetic estimation pointed the novelty of the taxon, as justified herein. Extending this research, a water-soluble polysaccharide-rich fraction was isolated to determine therapeutic prospects. Chemical characterization revealed that the backbone of the polymers, organized in triple-helical form, predominantly consisted of ß-glucan accompanied by a lower extent of galactose, mannose and xylose. Subsequently, the effective antioxidant activity was noted in terms of radical scavenging, reducing power and chelating ability with EC50 of 305-2726 µg ml-1. Further, the macromolecules triggered murine macrophages to proliferate, phagocytose, release NO, produce intracellular ROS and change morphodynamics. A significant alleviation in the expression of TLR-2, TLR-4, NF-κB, COX-2, TNF-α, Iκ-Bα, IFN-γ, IL-10 and iNOS was also observed explaining the definite immune-stimulatory activity and supporting traditional consumption of "Jam Patra" as a health-promoting food. Altogether, the study introduces a species in the world's myco-diversity and tribal food list opening doors of various opportunities in functional food and nature-based drug development arenas, which are currently in trend.

Mushroom data creation, curation, and simulation to support classification tasks.

Predicting if a set of mushrooms is edible or not corresponds to the task of classifying them into two groups-edible or poisonous-on the basis of a classification rule. To support this binary task, we have collected the largest and most comprehensive attribute based data available. In this work, we detail the creation, curation and simulation of a data set for binary classification. Thanks to natural language processing, the primary data are based on a text book for mushroom identification and contain 173 species from 23 families. While the secondary data comprise simulated or hypothetical entries that are structurally comparable to the 1987 data, it serves as pilot data for classification tasks. We evaluated different machine learning algorithms, namely, naive Bayes, logistic regression, and linear discriminant analysis (LDA), and random forests (RF). We found that the RF provided the best results with a five-fold Cross-Validation accuracy and F2-score of 1.0 ([Formula: see text], [Formula: see text]), respectively. The results of our pilot are conclusive and indicate that our data were not linearly separable. Unlike the 1987 data which showed good results using a linear decision boundary with the LDA. Our data set contains 23 families and is the largest available. We further provide a fully reproducible workflow and provide the data under the FAIR principles.

Pyrosequencing and Taxonomic Composition of the Fungal Community from Soil of Tricholoma matsutake in Gyeongju.

Tricholoma matsutake is an ectomycorrhizal fungus that has a symbiotic relationship with the root of Pinus densiflora. Soil microbial communities greatly affect the growth of T. matsutake, however, few studies have examined the characteristics of these communities. In the present study, we analyzed soil fungal communities from Gyeongju and Yeongdeok using metagenomic pyrosequencing to investigate differences in fungal species diversity, richness, and taxonomic composition between the soil under T. matsutake fruiting bodies (Sample 2) and soil where the fairy ring of T. matsutake was no longer present (Sample 1). The same spot was investigated three times at intervals of four months to observe changes in the community. In the samples from Yeongdeok, the number of valid reads was lower than that at Gyeongju. The operational taxonomic units of most Sample 2 groups were less than those of Sample 1 groups, indicating that fungal diversity was low in the T. matsutakedominant soil. The soil under the T. matsutake fruiting bodies was dominated by more than 51% T. matsutake. From fall to the following spring, the ratio of T. matsutake decreased. Basidiomycota was the dominant phylum in most samples. G-F1-2, G-F2-2, and Y-F1-2 had the genera Tricholoma, Umbelopsis, Oidiodendron, Sagenomella, Cladophialophora, and Phialocephala in common. G-F1-1, G-F2-1, and Y-F1-1 had 10 genera including Umbelopsis and Sagenomella in common. From fall to the following spring, the amount of phyla Basidiomycota and Mucoromycota gradually decreased but that of phylum Ascomycota increased. We suggest that the genus Umbelopsis is positively related to T. matsutake.

Phylogenetic analysis of the distribution of deadly amatoxins among the little brown mushrooms of the genus Galerina.

Some but not all of the species of 'little brown mushrooms' in the genus Galerina contain deadly amatoxins at concentrations equaling those in the death cap, Amanita phalloides. However, Galerina's ~300 species are notoriously difficult to identify by morphology, and the identity of toxin-containing specimens has not been verified with DNA barcode sequencing. This left open the question of which Galerina species contain toxins and which do not. We selected specimens for toxin analysis using a preliminary phylogeny of the fungal DNA barcode region, the ribosomal internal transcribed spacer (ITS) region. Using liquid chromatography/mass spectrometry, we analyzed amatoxins from 70 samples of Galerina and close relatives, collected in western British Columbia, Canada. To put the presence of toxins into a phylogenetic context, we included the 70 samples in maximum likelihood analyses of 438 taxa, using ITS, RNA polymerase II second largest subunit gene (RPB2), and nuclear large subunit ribosomal RNA (LSU) gene sequences. We sequenced barcode DNA from types where possible to aid with applications of names. We detected amatoxins only in the 24 samples of the G. marginata s.l. complex in the Naucoriopsis clade. We delimited 56 putative Galerina species using Automatic Barcode Gap Detection software. Phylogenetic analysis showed moderate to strong support for Galerina infrageneric clades Naucoriopsis, Galerina, Tubariopsis, and Sideroides. Mycenopsis appeared paraphyletic and included Gymnopilus. Amatoxins were not detected in 46 samples from Galerina clades outside of Naucoriopsis or from outgroups. Our data show significant quantities of toxin in all mushrooms tested from the G. marginata s.l. complex. DNA barcoding revealed consistent accuracy in morphology-based identification of specimens to G. marginata s.l. complex. Prompt and careful morphological identification of ingested G. marginata s.l. has the potential to improve patient outcomes by leading to fast and appropriate treatment.

Wild food plants and fungi sold in the markets of Luang Prabang, Lao PDR.

BACKGROUND: Open air markets hold an important position for ethnobiologists. In Southeast Asia, they are seriously understudied, in spite of their incredible biocultural diversity. In order to fill this gap we recorded plants and fungi sold in the open air markets of Luang Prabang, Lao PDR. METHODS: The markets were visited 38 times in four seasons: the dry season, early monsoon, mid-monsoon, and end-of-monsoon, at least 8 times per season. All items were photographed and voucher specimens were collected. Fungi were identified using DNA barcoding techniques. RESULTS: We recorded 110 species of wild edible plants and 54 species of fungi, including 49 wild-collected species. The sold plants included 86 species of green vegetables, 18 species of fruits and 3 species of flowers. Products from woody species constitute around half of all taxa sold. These include the young shoots of tree leaves, which are used for salads-an interesting feature of Lao cuisine. A large number of extremely rare Russula, with no reference sequences represented in databases or even species unknown to science is present on sale in the markets. CONCLUSIONS: Luang Prabang markets are some of the richest in species of wild edible plants and fungi in Asia, and indeed in the whole world. It is worth pointing out the exceptionally long list of wild edible mushrooms which are sold in Luang Prabang (and probably elsewhere in Laos). We view the Morning Market of Luang Prabang as a cultural treasure that unites the traditions of eating a large number of living species with very diverse flora and fauna. Measures should be taken to strike a balance between local foraging traditions and nature conservation priorities.

The genera Leucoagaricus and Leucocoprinus in the Dominican Republic.

We studied species of Leucoagaricus and Leucocoprinus collected in the Dominican Republic over the past 10 years using morphological and molecular methods and carefully compared our collections with previously described neotropical taxa. Twelve new species, eight in Leucoagaricus (La. bulbiger, La. caeruleovertens, La. margaritifer, La. pegleri, La. roseovertens, La. silvestris, La. stillatus, La. turgipes) and four in Leucocoprinus (Lc. antillarum, Lc. fuligineopunctatus, Lc. microlepis, Lc. scissus) are described. Additional records of previously described taxa are also discussed, including the first molecularly annotated occurrences of Lepiota guatopoensis, Lepiota mucrocystis, and La. rubroconfusus in their putative natural habitats and of Lc. cretaceus in the neotropics. Lepiota guatopoensis and Lepiota mucrocystis are transferred here to Leucoagaricus based on their phylogenetic placement and morphological characteristics. Color photographs of fresh basidiocarps and line drawings of microscopic characters are provided for all species.

Phlebopus roseus, a new edible bolete from China, is associated with insects and plants.

Phlebopus roseus is described as new based on collections from southwest China. Phylogenetic analyses of nuclear rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS) and portions of nuclear 28S rDNA (28S), translation elongation factor 1-alpha (tef1), and the largest and second largest subunits of RNA polymerase II (rpb1, rpb2) support P. roseus as a novel species in the genus Phlebopus (Boletinellaceae, Boletales). The new species resembles P. portentosus but differs from it in that mature basidiomata have a bright rose-red-colored stipe and a radiate tubular hymenophore with nested pores. Despite extensive searching, P. roseus has only been found at four sites within a 24-hectare orchard dominated by Eriobotrya japonica, which is agriculturally important given its fruit production (loquats). Therefore, this species appears to be endemic and geographically restricted. The ecology of this bolete is also unique. In line with the trophic behavior of other species in the Boletinellaceae, our observations indicate that P. roseus forms a symbiotic association with the scale insect Coccus hesperidum, identified through sequence analysis of its mitochondrial cytochrome c oxidase subunit I (COI) region, to form fungus-insect galls that develop on roots of E. japonica trees. Phlebopus roseus is an edible mushroom species and is collected from the type location by farmers and sold commercially in limited quantities at local markets alongside P. portentosus and other fungi.

A phylogenetic assessment of Pholiota and the new genus Pyrrhulomyces.

Multigene data sets were assembled to evaluate the phylogeny of species attributed to the genus Pholiota sensu A.H. Sm. & Hesler. This effort included generation of just more than 200 new sequences from 19 type collections of Pholiota and recent samples from East Asia. Phylogenetic analyses reinforced the autonomous phylogenetic positions of pholiotoid taxa in the genera Flammula (Hymenogastraceae) and Kuehneromyces (Strophariaceae). Samples of Pholiota astragalina from diverse geographic regions split into two species-level lineages but occupied an isolated phylogenetic position apart from Pholiota sensu stricto. The new genus Pyrrhulomyces is described to accommodate P. astragalina and a new cryptic species from the Southern Appalachians, Pyrrhulomyces amariceps. Pyrrhulomyces is distinguished from other genera of Strophariaceae by the blackening basidiomata with a bitter taste, smooth basidiospores without a germ pore under light microscopy, presence of pleurochrysocystidia, an ixocutis, rugulose spore ornamentation under scanning electron microscope (SEM), and association with late stages of conifer wood decay. Pholiota subochracea was found to be sister to a clade containing samples of Hypholoma and Bogbodia, but this portion of the Strophariaceae will require further taxon and gene sampling to resolve relationships between these three taxa. Pholiota sensu stricto comprised at least two major groups, but several residual poorly placed lineages were also noted depending on the data set analyzed. New combinations are made in the genera Flammula, Kuehneromyces, and Stropharia for three species of Pholiota-P. abieticola, P. obscura, and P. scabella, respectively, based on molecular annotation of type collections. Overall, 20 new synonymies are proposed, mostly in Pholiota. Illustrations of Pyrrhulomyces are provided along with a key to genera of Strophariaceae and Hymenogastraceae.

Amanita in the Guineo-Congolian rainforest: Epitypes and new species from the Dja Biosphere Reserve, Cameroon.

Four epitypes and three new species of Amanita (Amanitaceae, Agaricales, Agaricomycetes, Basidiomycota) are described from Guineo-Congolian rainforests of Cameroon. Amanita echinulata, A. fulvopulverulenta, A. robusta, and A. bingensis are epitypified based on collections that are the first since the species were described nearly a century ago. Morphological features of the epitypes are described and enumerated. Amanita minima, Amanita luteolamellata, and A. goossensfontanae are described as new and added to the known macromycota of tropical Africa. Habit, habitat, and known distribution are provided for each species. Sequence data for the internal transcribed spacer (ITS) locus are provided for types and other collections of all taxa, and a molecular phylogenetic analysis across the genus Amanita corroborates morphology-based infrageneric placement for each.

Stabilization-destabilization and redox properties of laccases from medicinal mushroom Ganoderma lucidum and human pathogen Yersinia enterocolitica.

Laccases or benzenediol oxygen oxidoreductases (EC 1.10.3.2) are polyphenol multicopper oxidases that are known for their structural and functional diversity in various life forms. In the present study, the molecular and physico-chemical properties (redox-potential and secondary structures) of fungal laccase isozymes (FLIs) isolated from a medicinal mushroom Ganoderma lucidum were analyzed and compared with those of the recombinant bacterial laccases (rLac) obtained from different Yersinia enterocolitica strains. It was revealed that the FLIs contained His-Cys-His as the most conserved residue in its domain I Cu site, while the fourth and fifth residues were variable (Ile, Leu, or Phe). Evidently, the cyclic voltammetric measurements of Glac L2 at Type 1 Cu site revealed greater E° for ABTS/ABTS+ (0.312 V) and ABTS+/ABTS2+ (0.773 V) compared to the E° of rLac. Furthermore, circular dichroism-based conformational analysis revealed structural stability of the FLIs at acidic pH (3.0) and low temperature (<30 °C), while the isozymes were destabilized at neutral pH (7.0) and high-temperature conditions (>70 °C). The zymographic studies further confirmed the functional inactivation of FLIs at high temperatures (≥70 °C), predominantly due to domain unfolding. These findings provide novel insight into the evolution of the catalytic efficiency and redox properties of the FLIs, contributing to the existing knowledge regarding stress responses, metabolite production, and the biotechnological utilization of metabolites.