Biosynthesis, Gene Expression, and Pharmacological Properties of Triterpenoids of Ganoderma Species (Agaricomycetes): A Review.

For more than 6 millennia, Ganoderma species have been used in traditional Asian medicine due to their health benefits. Ganoderma synthesizes several compounds with biological activity, including lanostane-type triterpenoids like ganoderic acids (GAs), lucidones, and colosolactones. These triterpenoids have been investigated for their antiviral, hypoglycemic, and anticancer effects. GAs are highly oxygenated triterpenoids with different functional groups attached to lanostane skeleton. Their great chemical diversity makes GAs prospects for the development of new drugs to treat multiple illnesses such as cancer. The effect of GAs against cancer cells has been associated with their capability to inhibit specific targets such as STAT3, to induce apoptosis and cell cycle blockage, and to increase natural killer cell activity. Due to the biological activity of these molecules, novel strategies are being developed for Ganoderma production mainly by liquid cultivation, gene overexpression (HMGR, SQS, LS) by elicitors, and modified growing conditions (carbon and nitrogen sources, pH, temperature), which induce reactive oxygen species production, key compounds for secondary metabolism. In addition, some transcription factors are mainly expressed under stress conditions, such as cytochrome P450 genes, which participate in the regulation of triterpenoid synthesis. The fermentation process has been scaled up to a 300-L bioreactor, which shows good GA production. This article reviews current knowledge on bioactive triterpenoids of Ganoderma and their production, biosynthesis, and pharmacological properties, emphasizing gene expression in liquid culture. It also discusses the lack of information regarding other species with high potential.

Phenolic compounds of Phellinus spp. with antibacterial and antiviral activities.

Phellinus Quél is one of the largest genera of Hymenochaetaceae; it comprises about 220 species widely distributed on Earth. Most Phellinus species are lignicolous mushrooms that accumulate bioactive compounds. This research studied the phenolic composition of Phellinus spp. and their relationship with antibacterial and antiviral capacity. Phenolics were extracted from Phellinus badius, P. fastuosus, and P. grenadensis; their antiviral and antibacterial activities were evaluated against Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica, and Escherichia coli O157: H7; and the bacteriophages MS2 and Φ- × 174. Gallic acid, chlorogenic acid, caffeic acid, epicatechin, ferulic acid, catechin, 1,3-dicaffeoylquinic acid, p-coumaric acid, and rutin were found in different proportions among Phellinus spp. Total phenolic content ranged from 96 to 209 mg GAE/g, and total flavonoids from 10 to 27 QE/g. The minimum inhibitory concentrations of P. badius, P. grenadensis, and P. fastuosus against E. coli O157: H7 were 13, 20, and 27 mg/mL, against S. enterica were 20, 30, and 15 mg/mL, and against L. monocytogenes were 10, 15, and 25 mg/mL, respectively. The phenolic content was better correlated with the antibacterial effect against E. coli O157: H7 and L. monocytogenes (r = 0.8-0.9), but not against S. enterica (r = 0.05). The antiviral activity of the extracts (0.9 mg/mL) was 29 to 41% against MS2 and 27 to 38% for Φ-X174 virus (r = 0.8-0.9). In silico analysis showed binding energy values of - 7.9 and - 4.8 kcal/mol between the identified phenolic compounds and the M and G proteins of each virus. The antibacterial and antiviral properties of Phellinus species were correlated with the phenolic content.

Contribution of Bioactive Compounds to the Antioxidant Capacity of the Edible Mushroom Neolentinus lepideus.

Neolentinus lepideus is a fungus consumed by rural communities in Central America and Asia due to its rich flavor; however, little information on its chemical composition is available. With this in mind, the objective of this work was to determine the content of vitamin E and C, ergosterol, and phenolic compounds of this fungus, as well as its antioxidant capacity. The quantified bioactive compounds were two isoforms of vitamin E, highlighting α-tocopherol (3370.35 mg/100 g dry weight, DW) and ergosterol (11.70 mg/100 g DW). The total phenolic content was 164.80 mg gallic acid equivalents/100 g, and nine phenolic compounds were identified (protocatechuic, p-hydroxybenzoic, caffeic, vanillic, ferulic, salicylic, p-anisic, trans-cinnamic acids, and scopoletin). The highest antioxidant capacity was detected in the lipophilic extract with TEAC (27688 µmoles Trolox equivalents/100 g). These results suggest that lipophilic compounds are among the main bioactive compounds in N. lepideus, and they might exhibit the highest radical scavenging properties in non-polar extracts.

Antioxidant Properties and Industrial Uses of Edible Polyporales.

The content of antioxidant compounds varies within fungal species, and the Polyporales order has been recognized for this property. Numerous antioxidant compounds have been identified in Polyporales fungi, including phenolic compounds, ß-glucans, ergosterol, ergothioneine, vitamin C, and tocopherols. Each compound contributes differently to the antioxidant potential of fungi. Besides the health benefits for rural communities caused by fungi consumption, their antioxidant composition attracts the food, cosmetic, and pharmaceutical industries' interest. In this context, the present review compiles, analyzes, and discusses the bioactive composition of edible fungi of the Polyporales order and its contribution to total antioxidant capacity.

A novel sequestrate species from Mexico: Aroramyces guanajuatensis sp. nov. (Hysterangiaceae, Hysterangiales).

Knowledge of sequestrate Hysterangiaceae fungi in Mexico is very limited. In the present study, a new member of the family, Aroramyces guanajuatensis sp. nov., is described. This speciesis closely related to A. balanosporus, but differs from the latter in possessing a tomentose peridium 165-240 µm thick, with occasional large terminal hyphae up to 170 µm, a variable mesocutis (isodiametric to angular), and distinct bright yellowish subcutis. In contrast, A. balanosporus possesses a fibrillose peridial surface with shorter hyphae, a peridium 200-450 µm thick, and a mainly hyaline isodiametric mesocutis with a slightly wider subcutis. The phylogenetic analysis of the LSU gene separated A. guanajuatensis from A. balanosporus with a Bayesian posterior probability (PP) = 1. This is the third Aroramyces species described for the American continent.

Review of Bioactive Molecules Production, Biomass, and Basidiomata of Shiitake Culinary-Medicinal Mushrooms, Lentinus edodes (Agaricomycetes).

Lentinus edodes (shiitake) is a basidiomycete that has been consumed for more than 2000 years because of its nutritional value and health benefits. It has a low lipid content, high fiber content, and a considerable amount of proteins; it also contains B vitamins and minerals in addition to a wide range of functional metabolites including polysaccharides, polysaccharopeptides, lectins, and secondary metabolites with bioactivity, e.g., lentinan, a ß-(1-3)-glucan with immunomodulatory activity, among others. Extracts and pure compounds of shiitake exhibit antibacterial, antifungal, cytostatic, antioxidant, anticancer, and immunomodulatory activity. Because of these attributes, different products derived from shiitake are on the market and are sold as dietary supplements. The traditional substrate for shiitake production is oak wood, yet the search for unconventional substrates has intensified over the past three decades. In particular, submerged cultivation of medicinal mushrooms has attracted great interest because it enables greater control of different fermentation factors to obtain products of interest. However, it is necessary to perform in vivo studies to determine the appropriate doses, side effects, and action spectrum of different bioactive compounds and fractions as well as to improve their production in liquid media and to potentiate their activity. We present an updated review of existing studies on the production of biomass and bioactive compounds of L. edodes in liquid culture and on solid fermentation for obtaining secondary mycelia and basidiomata.

Vineyard Pruning Waste Improves Bioconversion and Chemical Composition of Native Ganoderma spp. (Agaricomycetes) Strains from Mexico.

The aim of this work was to evaluate wood chips from vineyard prunings (VPs) as a potential substrate in cultures of Ganoderma spp. Biological efficiency (BE), production rate (PR), yield, and protein and fat contents of basidiomes increased when the wild strains G. oerstedii and G. subincrustatum, which were isolated from the Sonoran Desert, were cultivated on VPs. The mineral content of the basidiomes varied depending on the strain and substrate. The carbohydrate and phenol contents of the different substrate combinations and of the basidiomes were similar among strains. However, the carbohydrate and phenol contents of the substrates did not correlate with an increase in BE. Conversely, the high availability of protein, fat, and hemicellulose in VPs positively correlated with increases in BE, in yield, and in the protein and fat contents of the basidiomes of the wild strains. Our results clearly demonstrate that, in comparison with the traditional substrate of oak wood, VPs improve the production and biochemical composition of basidiomes. Therefore, cultivation of Ganoderma spp. on VPs is an environmentally friendly strategy for increasing their nutritional value and for cultivating these mushrooms for other biotechnological applications.

Productivity, Physicochemical Changes, and Antioxidant Activity of Shiitake Culinary-Medicinal Mushroom Lentinus edodes (Agaricomycetes) Cultivated on Lignocellulosic Residues.

The effects of substrate and strain on productivity, physicochemical characteristics, and compounds with antioxidant activity were evaluated in basidiomes of the shiitake mushroom, Lentinus edodes. Strains IE-245 and IE-256 and the substrates oak wood shavings (OW), sorghum stubble (SS), and sugar cane bagasse (SC) were used. Productivity was evaluated by measuring biological efficiency (BE), production rate (PR), and yield. Total sugars, total soluble solids, pH, titratable acidity, color parameters, total phenolics, flavonoids, ascorbic acid, and antioxidant activity of the basidiomes were measured. BE, PR and yield were higher with the combination IE-256/SS, at 103.71%, 1.32%, and 34.57%, respectively. The largest amount of total sugars (17.61 mg glucose · g-1 dry weight) was found with combination IE-256/SS. Variation was observed in basidiome color; the lowest luminosity (L*) value (darkest color) was found in the IE-256 strain on the OW substrate (L* = 30.45), whereas that of the IE-245 strain on the SC substrate was the lightest in color (L* = 57.00). The largest amounts of total phenolics were recorded in the IE-256 strain on the OW (6.50 mg gallic acid equivalents [GAE] · g-1 dry weight) and the SS substrates (5.85 mg GAE · g-1 dry weight). The best antioxidant activity was obtained with IE-256-0.80, 0.65, and 0.59 µmol Trolox equivalents · g dry weight-1-on the OW, SC, and SS substrates, respectively. Based on the values of BE, PR, and yield, IE-256/SS was the most productive. Substrate and strain, and their interactions, influenced the physicochemical characteristics of the basidiomes and the amounts of compounds with antioxidant activity they contained.

Response of ligninolytic macrofungi to the herbicide atrazine: dose-response bioassays / Respuesta de macrohongos ligninolíticos al herbicida atrazina: bioensayos dosis-respuesta

The effect of atrazine concentrations on mycelial growth and ligninolytic enzyme activities of eight native ligninolytic macrofungi isolated in Veracruz, México, were evaluated in a semi-solid culture medium. Inhibition of mycelial growth and growth rates were significantly affected (p = 0.05) by atrazine concentrations (468, 937, 1875, and 3750 mg/l). In accordance with the median effective concentration (EC50), Pleurotus sp. strain 1 proved to be the most tolerant isolate to atrazine (EC50 = 2281.0 mg/l), although its enzyme activity was not the highest. Pycnoporus sanguineus strain 2, Daedalea elegans and Trametes maxima showed high laccase activity (62.7, 31.9, 29.3 U mg/protein, respectively) without atrazine (control); however, this activity significantly increased (p < 0.05) (to 191.1, 83.5 and 120.6 U mg/protein, respectively) owing to the effect of atrazine (937 mg/l) in the culture medium. Pleurotus sp. strain 2 and Cymatoderma elegans significantly increased (p < 0.05) their manganese peroxidase (MnP) activities under atrazine stress at 468 mg/l. The isolates with high EC50 (Pleurotus sp. strain 1) and high enzymatic activity (P. sanguineus strain 2 and T. maxima) could be considered for future studies on atrazine mycodegradation. Furthermore, this study confirms that atrazine can increase laccase and MnP activities in ligninolytic macrofungi.

Se evaluó el efecto de diferentes concentraciones de atrazina sobre el crecimiento micelial y la actividad enzimática de ocho macrohongos ligninolíticos aislados en Veracruz, México. La inhibición del crecimiento micelial y la tasa de crecimiento diaria fueron significativamente (p < 0,05) afectadas por todas las dosis de atrazina (468, 937, 1875 y 3750 mg/l) adicionadas al medio de cultivo. De acuerdo con la concentración efectiva media (CE50), Pleurotus sp. cepa 1 fue el aislamiento más tolerante a la atrazina (CE50 = 2281 mg/l), aunque sus actividades enzimáticas no fueron altas. Pycnoporus sanguineus cepa 2, Daedalea elegans y Trametes maxima mostraron actividades altas de lacasa (62,7, 31,9 y 29,3 U mg/proteína, respectivamente) en ausencia de atrazina (control); estas actividades se incrementaron (p < 0,05) significativamente (191,1, 83,5 y 120,6 U mg/proteína, respectivamente) en presencia de atrazina (937 mg/l) en el medio de cultivo. Pleurotus sp. cepa 2 y Cymatoderma elegans incrementaron significativamente (p < 0,05) sus actividades de manganeso peroxidasa (MnP) bajo la concentración de 468 mg/l de atrazina. Los aislamientos con alta CE50 (Pleurotus sp. cepa 1) y alta actividad enzimática (P. sanguineus cepa 2 y T. maxima) podrían ser considerados para futuros estudios en la micodegradación de atrazina. Además, el presente estudio confirma que la atrazina puede incrementar las actividades lacasa y MnP en macrohongos ligninolíticos.

Response of ligninolytic macrofungi to the herbicide atrazine: dose-response bioassays / Respuesta de macrohongos ligninolíticos al herbicida atrazina: bioensayos dosis-respuesta

The effect of atrazine concentrations on mycelial growth and ligninolytic enzyme activities of eight native ligninolytic macrofungi isolated in Veracruz, México, were evaluated in a semi-solid culture medium. Inhibition of mycelial growth and growth rates were significantly affected (p = 0.05) by atrazine concentrations (468, 937, 1875, and 3750 mg/l). In accordance with the median effective concentration (EC50), Pleurotus sp. strain 1 proved to be the most tolerant isolate to atrazine (EC50 = 2281.0 mg/l), although its enzyme activity was not the highest. Pycnoporus sanguineus strain 2, Daedalea elegans and Trametes maxima showed high laccase activity (62.7, 31.9, 29.3 U mg/protein, respectively) without atrazine (control); however, this activity significantly increased (p < 0.05) (to 191.1, 83.5 and 120.6 U mg/protein, respectively) owing to the effect of atrazine (937 mg/l) in the culture medium. Pleurotus sp. strain 2 and Cymatoderma elegans significantly increased (p < 0.05) their manganese peroxidase (MnP) activities under atrazine stress at 468 mg/l. The isolates with high EC50 (Pleurotus sp. strain 1) and high enzymatic activity (P. sanguineus strain 2 and T. maxima) could be considered for future studies on atrazine mycodegradation. Furthermore, this study confirms that atrazine can increase laccase and MnP activities in ligninolytic macrofungi.(AU)

Se evaluó el efecto de diferentes concentraciones de atrazina sobre el crecimiento micelial y la actividad enzimática de ocho macrohongos ligninolíticos aislados en Veracruz, México. La inhibición del crecimiento micelial y la tasa de crecimiento diaria fueron significativamente (p < 0,05) afectadas por todas las dosis de atrazina (468, 937, 1875 y 3750 mg/l) adicionadas al medio de cultivo. De acuerdo con la concentración efectiva media (CE50), Pleurotus sp. cepa 1 fue el aislamiento más tolerante a la atrazina (CE50 = 2281 mg/l), aunque sus actividades enzimáticas no fueron altas. Pycnoporus sanguineus cepa 2, Daedalea elegans y Trametes maxima mostraron actividades altas de lacasa (62,7, 31,9 y 29,3 U mg/proteína, respectivamente) en ausencia de atrazina (control); estas actividades se incrementaron (p < 0,05) significativamente (191,1, 83,5 y 120,6 U mg/proteína, respectivamente) en presencia de atrazina (937 mg/l) en el medio de cultivo. Pleurotus sp. cepa 2 y Cymatoderma elegans incrementaron significativamente (p < 0,05) sus actividades de manganeso peroxidasa (MnP) bajo la concentración de 468 mg/l de atrazina. Los aislamientos con alta CE50 (Pleurotus sp. cepa 1) y alta actividad enzimática (P. sanguineus cepa 2 y T. maxima) podrían ser considerados para futuros estudios en la micodegradación de atrazina. Además, el presente estudio confirma que la atrazina puede incrementar las actividades lacasa y MnP en macrohongos ligninolíticos.(AU)

Improvement of yield of the edible and medicinal mushroom Lentinula edodes on wheat straw by use of supplemented spawn.

The research evaluated the interactions of two main factors (strain / types of spawn) on various parameters with the purpose to assess its effect on yield and biochemical composition of Lentinula edodes fruiting bodies cultivated on pasteurized wheat straw. The evaluation was made with four strains (IE-40, IE-105, IE-124 and IE-256). Different types of spawns were prepared: Control (C) (millet seed, 100%), F1 (millet seed, 88.5%; wheat bran, 8.8%; peat moss, 1.3%; and CaS04, 1.3%) and F2 (the same formula as F1, but substituting the wheat bran with powdered wheat straw). Wheat straw was pasteurized by soaking it for 1 h in water heated to 65 °C. After this the substrate (2 kg wet weight) was placed in polypropylene bags. The bags were inoculated with each spawn (5% w/w) and incubated in a dark room at 25 °C. A proximate analysis of mature fruiting bodies was conducted. The mean Biological Efficiency (BE) varied between 66.0% (C-IE-256) and 320.1% (F1-IE-124), with an average per strain of 125.6%. The highest mean BE was observed on spawn F1 (188.3%), significantly different from C and F2. The protein content of fruiting bodies was high, particularly in strain IE-40-F1 (17.7%). The amount of fat varied from 1.1 (F1-IE-40) to 2.1% (F2-IE-105) on dry matter. Carbohydrates ranged from 58.8% (F1-IE-40) to 66.1% (F1-IE-256). The energy value determined ranged from 302.9 kcal (F1-IE-40) to 332.0 kcal (F1-IE-256). The variability on BE observed in this study was significantly influenced by the spawn's formulation and genetic factors of the different strains.

Improvement of yield of the edible and medicinal mushroom Lentinula edodes on wheat straw by use of supplemented spawn

The research evaluated the interactions of two main factors (strain / types of spawn) on various parameters with the purpose to assess its effect on yield and biochemical composition of Lentinula edodes fruiting bodies cultivated on pasteurized wheat straw. The evaluation was made with four strains (IE-40, IE-105, IE-124 and IE-256). Different types of spawns were prepared: Control (C) (millet seed, 100%), F1 (millet seed, 88.5%; wheat bran, 8.8%; peat moss, 1.3%; and CaS0(4), 1.3%) and F2 (the same formula as F1, but substituting the wheat bran with powdered wheat straw). Wheat straw was pasteurized by soaking it for 1 h in water heated to 65 °C. After this the substrate (2 kg wet weight) was placed in polypropylene bags. The bags were inoculated with each spawn (5% w/w) and incubated in a dark room at 25 °C. A proximate analysis of mature fruiting bodies was conducted. The mean Biological Efficiency (BE) varied between 66.0% (C-IE-256) and 320.1% (F1-IE-124), with an average per strain of 125.6%. The highest mean BE was observed on spawn F1 (188.3%), significantly different from C and F2. The protein content of fruiting bodies was high, particularly in strain IE-40-F1 (17.7%). The amount of fat varied from 1.1 (F1-IE-40) to 2.1% (F2-IE-105) on dry matter. Carbohydrates ranged from 58.8% (F1-IE-40) to 66.1% (F1-IE-256). The energy value determined ranged from 302.9 kcal (F1-IE-40) to 332.0 kcal (F1-IE-256). The variability on BE observed in this study was significantly influenced by the spawn's formulation and genetic factors of the different strains.

Quantitative changes in the biochemical composition of lignocellulosic residues during the vegetative growth of Lentinula edodes

The chemical changes in barley-straw (BS), wheat-straw (WS) and vineyard-pruning (VP) substrates were determined during colonization of Lentinula edodes mycelia (during primordium development) in solid state fermentation. Primordia appeared 39-50 days after inoculation. VP appeared to promote early sporophore initiation. The concentration of hemicellulose in BS and VP decreased gradually from 25.5 percent to 15.6 percent and from 15.8 percent to 12.3 percent, respectively. However in WS, hemicellulose decreased from 27.2 percent to 9.5 percent. Lignin broke down continuously in BS and WS, with 31.8 percent and 34.4 percent degradation, respectively; higher than that of cellulose. During the pinning stage, the C:N ratio decreased in VP and BS, but not in WS. On all substrates the phenols decreased notably throughout the first week of mycelial growth. The time elapsed (days) to pinning was positively correlated with cellulose content (r=0.89), total sugar (r=0.85) and inversely correlated to lignin (r=-1.00) and phenol content (r=-0.55).

Quantitative changes in the biochemical composition of lignocellulosic residues during the vegetative growth of Lentinula edodes.

The chemical changes in barley-straw (BS), wheat-straw (WS) and vineyard-pruning (VP) substrates were determined during colonization of Lentinula edodes mycelia (during primordium development) in solid state fermentation. Primordia appeared 39-50 days after inoculation. VP appeared to promote early sporophore initiation. The concentration of hemicellulose in BS and VP decreased gradually from 25.5% to 15.6% and from 15.8% to 12.3%, respectively. However in WS, hemicellulose decreased from 27.2% to 9.5%. Lignin broke down continuously in BS and WS, with 31.8% and 34.4% degradation, respectively; higher than that of cellulose. During the pinning stage, the C:N ratio decreased in VP and BS, but not in WS. On all substrates the phenols decreased notably throughout the first week of mycelial growth. The time elapsed (days) to pinning was positively correlated with cellulose content (r=0.89), total sugar (r=0.85) and inversely correlated to lignin (r=-1.00) and phenol content (r=-0.55).

Bioconversion of agrowastes by Lentinula edodes: the high potential of viticulture residues.

The production of four strains of edible mushroom Lentinula edodes was evaluated through solid-state fermentation (SSF) of vineyard pruning (VP), barley straw (BS), and wheat straw (WS). Biological efficiency, proximal composition, and energy value of the fruiting bodies, as well as substrate chemical changes after harvest, were determined. The shortest primordium formation time (28 days), highest biological efficiency (93.25%), highest yield (37.46%), and shortest production cycle (6 days) were observed in VP. The fruiting bodies obtained from VP had high energy value (379.09 to 392.95 kcal) and contents of protein (12.37 to 17.19%), but low contents of fat (1.82 to 2.15%). After SSF, phenol concentration decreased on VP (1.2 mmol/L) and BS (0.31 mmol/L), but on WS remained practically the same. Hemicellulose decreased in all substrates; cellulose increased on WS and decreased in the rest of the treatments. Lignin decreased on WS and BS, but its concentration increased on VP. The variability observed in the degradation capacity of lignocellulosic components was influenced by the substrate's nature, environmental factors, and genetic factors among strains. VP has great potential for shiitake production due to its low cost, short production cycles, and high biological efficiency.

Evaluation of the viability of pleurotus spp. strains after liquid nitrogen cryopreservation

Viability of 6 mushrooms strains of the Pleurotus genus (2 from P. djamor var. djamor, 1 from P. ostreatus var. ostreatus, 2 from P. ostreatus var. columbinus and 1 from P. pulmonarius) after liquid nitrogen cryopreservation (-196§) was evaluated. The contact time for the mycelia of these strains with the cryoprotectant (glycerol) was studied 1,2 and 3 hours before freezing. We also tested the effect of different times (5, 10 and 15 minutes) and temperatures (30, 45 and 60§C) of the thawing system for mycelial recovery. The results showed a marked tendency toward faster mycelial recovery when samples were thawed at 30§C, while at 60§C no recovery was observed. A change in thawing and contact times with the cryoprotectant did not affect the results significantly, as the thawing temperature and strain employed affected